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Pubmed - Astaxantina

  • Astaxanthin: A Potential Therapeutic Agent in Cardiovascular Disease

    Robert G. Fassett 1,2,* and Jeff S. Coombes 2

    1    Renal Research Royal Brisbane and Women’s Hospital and The University of Queensland School of Medicine, Level 9 Ned Hanlon Building, Butterfield Street, Brisbane, Queensland 4029, Australia

    2    School of Human Movement Studies, The University of Queensland, St. Lucia, Brisbane,

    Queensland 4072, Australia; E-Mail: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.

     

    * Author to whom correspondence should be addressed; E-Mail: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.; Tel.: +61-419399571; Fax: +61-736368572.

     

    Received: 7 February 2011; in revised form: 14 March 2011 / Accepted: 18 March 2011 /

    Published: 21 March 2011

         

     

     

    Abstract: Astaxanthin is a xanthophyll carotenoid present in microalgae, fungi, complex plants, seafood, flamingos and quail. It is an antioxidant with anti-inflammatory properties and as such has potential as a therapeutic agent in atherosclerotic cardiovascular disease. Synthetic forms of astaxanthin have been manufactured. The safety, bioavailability and effects of astaxanthin on oxidative stress and inflammation that have relevance to the pathophysiology of atherosclerotic cardiovascular disease, have been assessed in a small number of clinical studies. No adverse events have been reported and there is evidence of a reduction in biomarkers of oxidative stress and inflammation with astaxanthin administration. Experimental studies in several species using an ischaemia-reperfusion myocardial model demonstrated that astaxanthin protects the myocardium when administered both orally or intravenously prior to the induction of the ischaemic event. At this stage we do not know whether astaxanthin is of benefit when administered after a cardiovascular event and no clinical cardiovascular studies in humans have been completed and/or reported. Cardiovascular clinical trials are warranted based on the physicochemical and antioxidant properties, the safety profile and preliminary experimental cardiovascular studies of astaxanthin.

     

    Keywords: antioxidants; xanthophyll carotenoid; inflammation; Haematococcus pluvialis; oxidative stress

     
       

     

     

     

    1. Introduction

     

    Astaxanthin is a xanthophyll carotenoid of predominantly marine origin, with potent antioxidant  and anti-inflammatory effects demonstrated in both experimental and human studies. Oxidative stress and inflammation are common pathophysiological features of atherosclerotic cardiovascular disease hence astaxanthin may have a potential therapeutic role in this condition. This review will summarise the available evidence suggesting astaxanthin may be of therapeutic value in cardiovascular disease.

     

    2.   Oxidative Stress and Inflammation

     

    Oxidative stress and inflammation are established non-traditional risk factors for atherosclerosis associated cardiovascular morbidity and mortality [1]. Dietary antioxidants can reduce the oxidation of lipids and proteins and have the potential to protect from the development of arterial stiffening and atherosclerosis [2–4]. Cross-sectional and prospective observational studies have demonstrated an association between the intake of dietary antioxidants and/or their plasma levels and a reduction of cardiovascular events [5–10]. This supports the theory that oxidative stress is a pathophysiological process involved in atherosclerotic vascular damage. Also, a reduced dietary antioxidant intake is associated with oxidative stress and inflammation [11]. Newer more potent dietary antioxidants such   as astaxanthin have yet to be studied in this setting. Studies that have assessed the intake of b-carotene or dietary b-carotene supplementation have shown higher b-carotene consumption is associated with a reduction in cardiovascular disease [6,12–17]. Other than a few studies [18–20], cardiovascular intervention trials using antioxidants have not demonstrated benefits [21–23]. This may be because study participants did not have oxidative stress and/or the antioxidants used were insufficiently potent. In addition, it is becoming recognized that there is communication between oxidative stress and inflammatory processes leading to the additional hypothesis that antioxidants may be able to modify both deleterious events. Further research is needed studying antioxidants with different biological actions in patients with demonstrated oxidative stress.

     

    3.   Carotenoids

     

    Carotenoids are ubiquitous, and found in high concentrations in plants, algae and microorganisms. Humans and other animals cannot synthesize them and therefore are required to source them in        their diet [24]. Carotenoids are classified, according to their chemical structure, into carotenes and xanthophylls. The carotene carotenoids include b-carotene and lycopene and the xanthophyll carotenoids include lutein, canthaxanthin, zeaxanthin, violaxanthin, capsorubin and astaxanthin [25,26].

    The effects of carotenoids vary dependent on how they interact with cell membranes [25]. The effects of astaxanthin, zeaxanthin, lutein, b-carotene and lycopene on lipid peroxidation have been assessed using a polyunsaturated fatty acid enriched membrane model [25,27]. Non-polar carotene carotenoids such as lycopene and b-carotene caused membrane disorder and lipid peroxidation in contrast to the polar xanthophyll carotenoid astaxanthin, which preserved membrane structure [27]. Contrasting effects of different carotenoids may be responsible for the differing biological effects seen in clinical studies. For instance, in some studies the non-polar carotenoid, b-carotene has been shown  to  have  no  benefit  on  cardiovascular  disease  [28–32]  and  in  fact  it  may  be  pro-oxidant  at high

     

     

    doses [33]. In contrast, the polar carotenoid astaxanthin has protective effects on the cardiovascular system demonstrated in animal studies. However,  this  has  not  been  studied  in  human  clinical  trials [34–36]. b-carotene at physiological levels may act in differing ways when ultraviolet A light A (UVA) acts on keratinocytes including vitamin A-independent pathways [37]. Astaxanthin, canthaxanthin  and  b-carotene  had  differential  effects  on  UVA-induced  oxidative  damage  [38].   In addition, carotenoids may also alter the immune response [39] and transcription [40].

     

    4.   Astaxanthin

     

    Astaxanthin contains two oxygenated groups on each ring structure (see Figure 1), which is responsible for its enhanced antioxidant features [41]. It is found in living organisms particularly in the marine environment where it is present in microalgae, plankton, krill and seafood. It gives salmon, trout, and crustaceans such as shrimp and  lobster  their  distinctive  reddish  coloration  [42].  It  is  also present in yeast, fungi, complex plants and the feathers of some birds including flamingos and quail [42]. In 1987, the United States Food and Drug Administration approved astaxanthin as a feed additive for use in the aquaculture industry and in 1999 it was approved for use as a  dietary  supplement (nutraceutical) [41]. The microalgae Haematococcus pluvialis produces the astaxanthin isomer (3S, 3S′), which is the same as the form found in wild salmon. Synthesis of astaxanthin is not possible in humans and it cannot be converted to vitamin A, which means excess intake will not cause hypervitaminosis A toxicity [43,44]. Astaxanthin and canthaxanthin are scavengers of free radicals, chain-breaking antioxidants and potent quenchers of reactive oxygen and nitrogen species including singlet oxygen, single and two electron oxidants [45–47]. They (astaxanthin and canthaxanthin) have terminal carbonyl groups that are conjugated to a polyene backbone [26] and are more potent antioxidants and scavengers of free radicals than carotene carotenoids such as b-carotene [47,48]. For these reasons dietary supplementation with astaxanthin has the potential to provide antioxidant protection of cells and from atherosclerotic cardiovascular disease [49].

     

    Figure 1. Molecular structure of astaxanthin.

     
       

     

     

    5.   Astaxanthin Formulations

     

    • Astaxanthin of Marine Origin

     

    Astaxanthin used in nutritional supplements is usually a mixture of configurational isomers produced by Haematococcus pluvialis, a unicellular microalga [50]. Astaxanthin can be produced       in its natural forms on a large scale [51].  The  initial  production  of  astaxanthin  from  Haematococcus pluvialis  uses  closed  culture  systems  followed  by  a  5–7  day,  “reddening”  cycle,

     

     

    conducted in open culture ponds. At each production stage, the cultures are closely monitored by microscopic examination to ensure they remain free of contamination. After the reddening cycle, Haematococcus pluvialis cultures are harvested, washed and dried. The final step for the production of astaxanthin is extraction of dried Haematococcus pluvialis biomass using supercritical carbon dioxide to produce a purified oleoresin, which is free of any contamination. Other sources used for the commercial production of astaxanthin include cultures of Euphausia pacifica (Pacific  krill),  Euphausia superba (Antarctic krill), Pandalus borealis (shrimp) and Xanthophyllomyces dendrorhous, formerly Phaffia rhodozyma (yeast). Astaxanthin from natural sources varies considerably from one organism to another. For instance, the astaxanthin found in seafood will depend on the stereoisomer ingested. Astaxanthin produced by haematococcus pluvialis, consists of the (3-S,3′-S) stereoisomer which is most commonly used in aquaculture. It is therefore the form most commonly consumed        by humans.

     

    • Synthetic Astaxanthin

     

    There are three stereoisomers of astaxanthin; (3-R,3′-R), (3-R,3′-S) and (3-S,3′-S). Disodium disuccinate astaxanthin (DDA) is a synthetic astaxanthin containing a mixture of all three stereoisomers, in the proportions 1:2:1. DDA was manufactured by Cardax Pharmaceuticals and used in animal studies investigating the myocardial ischemia-reperfusion injury models [34–36,52–54]. This form of astaxanthin was touted to have better aqueous solubility, unlike other carotenoids, and this enabled both oral and intravenous administration. DDA is no longer available but the same company now produces a second synthetic astaxanthin compound; Heptax/XanCor, CDX-085. The company claims that it is developed for thrombotic protection, triglyceride reduction, metabolic syndrome, and inflammatory liver disease. In addition, it has increased water dispersibility and enhanced bioavailability compared to natural astaxanthin and DDA. The synthetic forms are metabolized via hydrolysis in the intestine yielding free astaxanthin for intestinal absorption. CDX-085 has been used  in one study, discussed below [55].

    It is not yet clear which form of astaxanthin should be administered in clinical studies, the natural form from the marine environment or a synthetic form. As the proportions of stereoisomers, vary between these different forms of astaxanthin they may not be therapeutically equivalent [56]. Thus synthetic astaxanthin could result in different outcomes when assessed clinically [57].

     

    6.   Astaxanthin-Experimental Studies

     

    Experimental studies undertaken with astaxanthin specifically relevant to the cardiovascular system are summarised in Table 1. Astaxanthin attenuates mediators of oxidative stress and inflammation and has shown beneficial effects in non-cardiovascular models of disease [58–69]. In addition, astaxanthin has decreased markers of lipid peroxidation [70], inflammation [61,62,67,68] and thrombosis [55].

     

     

    Table 1. Animal studies investigating the cardiovascular effects of astaxanthin.

     

    Study

    Model

    Dosage

    Duration/timing of supplementation

    Effects of astaxanthin

    Lauver et al.

    2008 [34]

    Dog with occlusive carotid artery thrombus

    DDA 10, 30, or

    50 mg/kg/body weight IV

    30 min after occlusion

    - Reduced incidence of secondary thrombosis

    Aoi et al.

    2003 [63]

    C57BL/6 mice

    Diet supplemented with astaxanthin 0.02% weight/weight and food intake recorded

    3 weeks

    -   Attenuation of exercise increased 4-hydroxy-2- nonenal-modified protein and 8-hydroxy-2′- deoxyguanosine in cardiac and gastrocnemius muscle

    -   Attenuation of exercise increases in creatine kinase and myeloperoxidase activity in cardiac and gastrocnemius muscle

    -   Astaxanthin accumulated in cardiac and gastrocnemius muscle

    Gross and Lockwood 2004 [35]

    Myocardial infarct model Sprague-Dawley rats

    DDA 25/50/75 mg/kg body weight intravenously daily

    4 days prior to myocardial infarction

    - Myocardial infarct size significantly reduced

    Hussein et al.

    2005 [71]

    Stroke prone Spontaneously hypertensive rats

    Astaxanthin 50 mg/kg body weight/day

    5 weeks

    -   Significant blood pressure reduction

    -   Delayed incidence of stroke

    Lauver et al.

    2005 [52]

    Rabbit model of myocardial ischemia/reperfusion

    DDA 50 mg/kg body weight/day intravenously

    5 days

    -   Significant reduction in complement activation

    -   Significant reduction in myocardial infarct size

    Gross et al.

    2005 [54]

    Canine model of myocardial ischemia/reperfusion

    DDA 50 mg/kg body weight/day intravenously

    2 h or daily for four days

    -   Significant reduction in myocardial infarct size

    -   Two of three dogs treated for four days had 100% cardiac protection

     

     

    Table 1. Cont.

     

    Gross et al.

    2006 [36]

    Sprague-Dawley rats Left anterior descending coronary artery occlusion/reperfusion

    DDA 125 or 500 mg/kg body weight/day orally

    7 days

    -   Astaxanthin loading of myocardium indicating good bioavailability

    -   Trends in lowering of lipid peroxidation products

    -   Significant reduction in myocardial infarct size

    Hussein et al.

    2006 [72]

    Spontaneously hypertensive rats

    Astaxanthin 5% in olive oil (5 mg/kg/day orally)

    7 days

    -   Significant reduction in nitric oxide end products

    -   Significant reduction in elastin bands in aorta

    -   Significant reduction in wall/lumen arterial ratio in coronary arteries

    Aoi et al.

    2008 [73]

    ICR mice

    Astaxanthin 0.02% w/w

    4 weeks

    Astaxanthin increased fat utilization during exercise and prolonged exercise

    Astaxanthin prevented increase in hexanoyl-lysine modification of CPT I with exercise

    Nakao et al.

    2010 [74]

    BALC/c mice

    Astaxanthin 0, 0.02,

    0.08% orally/day

    8 weeks

    -   No change in blood glutathione concentration

    -   No change in lymphocyte mitochondrial membrane potential

    -   Higher myocardial mitochondrial membrane potential and contractility index

    Khan et al.

    2010 [55]

    C57BL/6 mice

     

     

    Human umbilical vein endothelial cells and platelets from

    Wistar-Kyoto rats

    CDX-085 500 mg/kg body weight/day

    14 days

    -   CDX-085 administered orally to C57BL/6 mice was associated with presence of free astaxanthin in the plasma, heart, liver and platelets

    -   Mice fed astaxanthin had significantly increased basal arterial blood flow and delay in occlusive thrombosis after endothelial injury

    -   Human umbilical vein endothelial cells and platelets from Wistar-Kyoto rats treated with free astaxanthin has significantly

                                                                                                                                            increased release of nitric oxide and decreased peroxynitrite levels

     

     

    • Cardiovascular Studies

     

    A series of experiments have been conducted to assess the efficacy of DDA in protecting the myocardium using the myocardial ischemia-reperfusion model in rats, rabbits and dogs [35,36,53,54]. Prior treatment for four-days with intravenous DDA using doses of 25, 50 and 75 mg/kg body weight in Sprague-Dawley rats significantly reduced myocardial infarct size [35]. The degree of cardiac protection correlated with the dose of DDA administered. In a study in rabbits using a myocardial ischaemia-reperfusion model prior intravenous treatment with 50 mg/kg/day of DDA for four days resulted in a significant decrease in the size of the myocardial infarction and an improvement in myocardial salvage [52]. Animals treated with DDA had an attenuation of inflammation and complement activation suggesting there was a reduction in tissue inflammation [52]. In another study using a dog model intravenous DDA was administered daily for four-days prior to occlusion of the left anterior descending coronary artery or two hours prior to coronary artery occlusion [54]. After an hour of coronary occlusion and three hours of reperfusion there was a significant reduction in myocardial infarct size in the dogs treated with DDA. In the four-day treatment group, two out of three dogs had complete cardiac protection [54]. In a rat study, the effects of seven days of pre-treatment with oral DDA, 125 and 500 mg/kg/day on the concentrations of free astaxanthin in myocardial tissue [36]. The astaxanthin concentration in the myocardium was 400 nM after oral DDA at a dose of 125 mg/kg/day for seven-days and it was 1634 nM after 500 mg/kg/day. There was also a reduction of multiple lipid peroxidation products. The doses of DDA used in these experiments were quite high and at this stage it is not known whether such doses would be safe to use in humans.

    The effects of astaxanthin  on blood pressure (BP) were assessed in spontaneously hypertensive   rats (SHR). There was a significant reduction in BP after 14-days of oral astaxanthin administration whereas this did not occur in normotensive Wistar Kyoto rats [71]. Astaxanthin administered orally for five-weeks in stroke prone SHR also resulted in a significant BP reduction [71]. Oral astaxanthin also enhanced nitric oxide induced vascular relaxation in the rat aortas [71] In experiments in SHR, oral astaxanthin significantly decreased nitric oxide end products indicating that it may be exerting its BP effects via this pathway [72]. Studies using the SHR aorta and coronary arteries demonstrated that astaxanthin reduced the wall/lumen ratio in coronary arteries  and  decreased  elastin  bands  in  the aorta [72]. This suggests that astaxanthin may beneficially mediate atherosclerotic CVD processes.

    Recently, a series of two experiments were reported in the one article, one using the synthetic astaxanthin (CDX-085) and the other using free astaxanthin [55]. CDX-085 administered orally to C57BL/6 mice resulted in the presence of free astaxanthin in the plasma, heart, liver and platelets.  Mice that were fed astaxanthin had significantly increased basal arterial blood flow and a delay in occlusive thrombosis after endothelial injury. Also, in an in vitro study, human umbilical vein endothelial cells and platelets isolated from Wistar-Kyoto rats that were treated with free astaxanthin has significantly increased nitric oxide release and a decrease in peroxynitrite levels [55]. The authors concluded the results support the potential of astaxanthin as a potential therapy to prevent thrombosis associated with cardiovascular disease.

    Astaxanthin administered to C57BL/6 mice resulted in a reduction in exercise-induced increases in the oxidative stress biomarkers 8-hydroxy-2′-deoxyguanosine and 4-hydroxy-2-nonenal-modified protein in both cardiac and gastrocnemius muscle [63]. Increases in myeloperoxidase and creatinine

     

     

    kinase activity in cardiac and gastrocnemius muscle were also reduced by astaxanthin. After three-weeks of astaxanthin supplementation there was evidence of accumulation of astaxanthin in gastrocnemius and cardiac muscle. Astaxanthin given to female BALB/c mice for eight-weeks resulted in a dose dependent increase in plasma astaxanthin but no significant changes in blood glutathione or change in lymphocyte mitochondrial membrane potential and cardiac contractility index measured on echocardiography. The mice that were fed 0.08% astaxanthin in the diet had higher cardiac mitochondrial membrane potential and contractility index compared with control animals [74]. This suggests dietary astaxanthin provides cardiac protection. Astaxanthin administered for four weeks to eight week old ICR mice resulted in increased exercised induced fat utilization and prevention of increased hexanoyl-lysine modification of carnitine palmitoyltransferase I (CTP I) [73]. In a canine carotid artery thrombosis model, administration of DDA resulted in a dose-dependent reduction in carotid artery re-thrombosis and a reduction of re-thrombosis after thrombolysis but there was no  effect on hemostasis [34].

     

    • Diabetes Studies

     

    Diabetes mellitus and its associated nephropathy is a common cause of chronic kidney disease and is complicated by accelerated atherosclerotic cardiovascular disease [75]. In studies involving diabetic db/db mice, supplementation with astaxanthin produced a reduction in the levels of blood glucose [60]. In the kidney there was significantly decreased relative mesangial area in the animals supplemented with astaxanthin. Also proteinuria and urinary excretion of 8-OHdG were attenuated. Mice supplemented with astaxanthin had less glomerular 8-OHdG immunoreactive cells [60]. Hyperglycemia induced reactive oxygen species production, activation of transcription factors, and cytokine expression and production by normal human mesangial cells was suppressed significantly by astaxanthin [66].

     

    7.   Astaxanthin Studies in Humans

     

    Although no cardiovascular outcomes studies using astaxanthin have been reported in humans there have been clinical studies that have investigated the effects of astaxanthin in human health and other diseases (Table 2). The majority of these have been conducted in healthy participants who volunteered to assess astaxanthin dose, bioavailability, safety and oxidative stress, which are all potentially relevant to the cardiovascular system. Studies have also been conducted in other medical conditions such as reflux oesophagitis, where measurements of oxidative stress and/or inflammation have been included.

     

    • Dosing

     

    Human clinical studies have  used  oral  astaxanthin  in  a  dose  that  ranges  from  4  mg  up  to  100 mg/day, given from a one off dose up to durations of one-year (Table 2).

     

     

    Table 2. Clinical studies investigating the safety, bioavailability and effects of astaxanthin on oxidative stress.

     

    Study

    Study  population (n = subject numbers)

    Dosage

    Study design

    Duration of supplementation

    Effects of astaxanthin

    Iwamoto et al.

    2000 [70]

    Volunteers (n = 24)

    Different doses: 1.8,

    3.6, 14.4,

    21.6 mg/day

    Open labelled

    2 weeks

    - Reduction of LDL oxidation

    Osterlie et al.

    2000 [76]

    Middle aged male volunteers (n = 3)

    100 mg

    Open labelled

    Single dose

    - Astaxanthin taken up by VLDL chylomicrons

    Mercke Odeberg et al.

    2003 [77]

    Healthy male volunteers (n = 32)

    40 mg

    Open labelled parallel

    Single dose

    - Enhanced bioavailability with lipid based formulation

    Spiller et al.

    2003 [78]

    Healthy adults (n = 35)

    6 mg/day

    (3 × 2 mg tablets/day)

    Randomised, double blind, placebo controlled

    8 weeks

    - Demonstrated safety assessed by measures of blood pressure and biochemistry

    Coral-Hinostroza et al.

    2005 [79]

    Healthy adult males (n = 3)

    10 mg and

    100 mg

    Open labelled

    Single dose or 4 weeks

    - Cmax 0.28 mg/L at 11.5 h at high dose and

    0.08 mg/L at low dose

    -   Elimination half life 52 ± 40 h

    -   z-isomer selectively absorbed

    Karppi et al.

    2007 [80]

    Healthy non-smoking Finnish males (n = 40)

    8 mg/day

    Randomised, double blind, placebo controlled

    12 weeks

    -   Intestinal absorption adequate with capsules

    -   Reduced levels of plasma 12 and 15 hydroxy fatty acids

    -   Decreased oxidation of fatty acids

     

     

    Table 2. Cont.

     

    Parisi et al.

    2008 [81]

    Non-advanced age related macular degeneration

    (n = 27)

    4 mg/day

    Randomised controlled trial open labelled no placebo

    12 months

    - Improved central retinal dysfunction in age related macular degeneration when administered with other antioxidants

    Miyawaki et al.

    2008 [82]

    Healthy males (n = 20)

    6 mg/day

    Single blind, placebo controlled

    10 days

    - Decreased whole blood transit time (improved blood rheology)

    Rufer et al.

    2008 [83]

    Healthy males (n = 28)

    5 mg/g salmon flesh (wild vs. aquacultured)

    Randomised, double blind, placebo controlled

    4 weeks

    - Bioavailability initially better with ingestion of aquacultured salmon but equivalent at day 28.

    Isomer (3S, 3′S) greater in plasma compared with isomer proportion in salmon flesh

    Park et al.

    2010 [84]

    Healthy females (n = 14)

    0, 2, 8 mg/day

    Randomised, double blind, placebo controlled

    8 weeks

    -   Decreased plasma 8-hydroxy-2′-deoxyguanosine after week four in those taking astaxanthin.

    -   Lower CRP after week four in those taking 2 mg/day astaxanthin

     

     

    • Bioavailability

     

    Astaxanthin bioavailability from the marine environment was assessed in a randomised double  blind trial in 28 volunteers [83]. Participants were given either 250 g of wild salmon or aquaculture salmon (5 µg/g) to eat. Wild salmon ingest astaxanthin naturally from krill whereas aquacultured salmon acquire it from fish that are fed astaxanthin that might be derived from a synthetic source. Plasma levels of astaxanthin were higher at 3, 6, 10 and 14 days during ingestion of the aquacultured compared with the wild salmon. Plasma levels of the (3-S, 3′-S) isomer of astaxanthin appeared at higher levels than its proportionate level in the flesh of the salmon. This suggests that isomers of astaxanthin might have different bioavailability. The plasma isomers of astaxanthin have also been studied after ingestion of single oral dose of 10mg and also 100 mg over four-weeks. Astaxanthin plasma elimination half-life was 52 (SD 40) h and there was a non-linear dose response and selective absorption of z-isomers [79].

     

    • Safety

     

    The safety of astaxanthin administered orally was assessed in a double-blind, randomised placebo-controlled trial undertaken in healthy adults [78]. Volunteers took either 6 mg/day of astaxanthin or placebo for eight-weeks. BP and biochemistry measured after four and eight weeks of therapy revealed no significant differences in these parameters between treatment and placebo groups and these did not differ from baseline. The authors concluded that healthy adults could safely consume 6 mg/day of astaxanthin derived from a Haematococcus pluvialis algal extract. Measuring whole blood transit time in 20 healthy males was used to assess the effects of astaxanthin on blood rheology in humans. Six milligrams of oral astaxanthin per day for ten days improved blood rheology as evidenced by decreased whole blood transit time [82]. Escalating concentrations of astaxanthin were tested          in vitro with blood taken from volunteers, 8 of whom were taking asprin and 12 who were not [85]. Even supra-therapeutic concentrations of astaxanthin had no adverse effects on indices of platelet, coagulation and fibrinolytic function. These results support the safety profile of astaxanthin for future clinical trials. No significant side effects have been reported so far in published human studies in  which astaxanthin was administered to humans.

     

    • Oxidative Stress and Inflammation

     

    Oral supplementation with astaxanthin in studies in healthy human volunteers and patients with reflux oesophagitis demonstrated a significant reduction in oxidative stress, hyperlipidemia and biomarkers of inflammation [70,80,86]. In a study involving 24 healthy volunteers who ingested astaxanthin in doses from 1.8 to 21.6 mg/day for two weeks, LDL lag time, as a measure of susceptibility of LDL to oxidation, was significantly greater in astaxanthin treated participants indicating inhibition of the oxidation of LDL [70]. Plasma levels of 12- and 15-hydroxy fatty acids were significantly reduced in 40 healthy non-smoking Finnish males given astaxanthin [80] suggesting astaxanthin decreased the oxidation of fatty acids [80]. The effects of dietary astaxanthin in doses of   0, 2 or 8 mg/day, over 8 weeks, on oxidative stress and inflammation were investigated in a double blind study in 14 healthy females [84]. Although these participants did not have oxidative stress or

     

     

    inflammation those taking 2 mg/day had lower CRP at week eight. There was also a decrease in DNA damage measured using plasma 8-hydroxy-2′-deoxyguanosine after week four in those taking astaxanthin. Astaxanthin therefore appears safe, bioavailable when given orally and is suitable for further investigation in humans.

     

    8.   Clinical Trial Using Astaxanthin

     

    A double-blind randomised placebo-controlled clinical trial (Xanthin study) is currently being conducted to assess the effects of astaxanthin 8mg orally day on oxidative stress, inflammation and vascular function in patients that have received a kidney transplant [87]. Patients in the study  undertake measurements of surrogate markers of cardiovascular disease including aortic pulse wave velocity, augmentation index, brachial forearm reactivity and carotid artery intima-media thickness. Depending on the results from this pilot study a large randomised controlled trial assessing major cardiovascular outcomes such as myocardial infarction and death may be warranted.

     

    9.   Conclusions

     

    Experimental evidence suggests astaxanthin may have protective effects on cardiovascular disease when administered prior to an induced ischemia-reperfusion event. In addition, there is evidence that astaxanthin may decrease oxidative stress and inflammation which are known accompaniments of cardiovascular disease. At this stage we do not know whether astaxanthin has any therapeutic value in human cardiovascular disease either in a preventative capacity or when administered after a cardiovascular insult. It has been proposed that astaxanthin may provide cardiovascular protection through reducing oxidative stress, which is one of the non-traditional risk factors for the development of atherosclerotic cardiovascular disease. The role of oxidative stress in cardiovascular disease is supported by evidence from observational studies that have found associations between antioxidant intake, oxidative stress and cardiovascular outcomes. Despite this, clinical intervention studies using antioxidants including vitamin E, b-carotene and vitamin C, have not proved successful [22,23]. These intervention studies may have failed because of flawed design where patients were not included based on the presence of oxidative stress. Hence, many participants may not have been in a state of oxidative stress and able to benefit from antioxidant therapy. Also, in those participants where oxidative stress may have existed there was no way of assessing whether the therapy adequately corrected this. Thus, the antioxidants used such as vitamin E, b-carotene and vitamin C may not have been effective  because insufficient doses were used or an inadequate length of therapy followed to correct the oxidative stress. Some antioxidants such as b-carotene may be pro-oxidant at higher doses, which could have confounded study results.

    Astaxanthin is a potent antioxidant and based on its physicochemical properties and the results of preliminary experimental studies in ischaemia-reperfusion models of cardiovascular disease, it warrants consideration for testing in human clinical trials. There have been no safety concerns noted so far in human clinical studies where astaxanthin has been administered. As astaxanthin is a potent antioxidant and is associated with membrane preservation, it may protect against oxidative stress and inflammation and provide cardiovascular benefits.

     

     

    Acknowledgements

     

    Cyanotech the manufacturer of BioAstin, a proprietary brand of astaxanthin, is providing financial support and astaxanthin capsules and placebo for a clinical trial being conducted by the authors. The sponsor has no role in the study itself.

     

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    1. Fassett, G.; Healy, H.; Driver, R.; Robertson, I.K.; Geraghty, D.P.; Sharman, J.E.; Coombes, J.S. Astaxanthin vs. placebo on arterial stiffness, oxidative stress and inflammation in renal transplant patients (Xanthin): A randomised controlled trial. BMC Nephrol. 2008, 9, 17.

     

    Samples Availability: Available from the authors.

     

    © 2011 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).

  • Discovering the link between nutrition and skin aging

    Silke K. Schagen,1,† vasiliki A. Zampeli,1, 2,† evgenia Makrantonaki1,2 and Christos C. Zouboulis1,*

    1Departments of Dermatology, venereology, Allergology and immunology, Dessau Medical Center; Dessau, Germany; 2Laboratory for Biogerontology, Dermato-Pharmacology and Dermato-endocrinology; institute of Clinical Pharmacology and Toxicology; Charité Universitaetsmedizin Berlin; Berlin, Germany

    †These authors contributed equally to this  work.

    Keywords: nutrition, diet, ultraviolet protection, skin aging, antioxidants, fatty acids, flavonoids,  vitamins

    Abbreviations: 1,25(OH)2D3, 1,25-dihydroxy vitamin D3; CoQ10, coenzyme Q10; CR, caloric restriction; EFAs, essential fatty acids; EGCG, (-)-epigallocatechin-3-gallate; FoxO transcription factors, forkhead box class O transcription factor; GH, growth hormone; GTPs, green tea polyphenols; DHEAS, dehydroepiandrosterone sulphate; HRT, hormone replacement therapy;

    IGF-I, Insulin-like growth factor-I; IU, international unit; JNK, jun N-terminus kinase; mTORC1, mammalian target of rapamycin complex 1; MMP, matrix metalloproteinase; MST1, STE-like 20 protein kinase 1; ROS, reactive oxygen species; UL, upper intake levels; UV,  ultraviolet

    Introduction

    Beauty comes from the inside. The connection between nutri- tion and skin condition or rather the effect of nutrition on skin aging has been an interesting research field not only for scien- tists but also a common field of interest for humans throughout the years, from ancient times to nowadays. Skin aging consists   of two didactically independent, clinically and biologically, dis- tinct processes.1 The first is intrinsic skin aging, which represents chronological aging and affects skin in the same pattern it affects all internal organs.2 The second is extrinsic skin aging, which we view as aged skin and is the result of external factors and environ- mental influence, mainly chronic sun exposure and ultraviolet (UV) irradiation but also smoking, pollution, sleep deprivation and poor nutrition.

    Prevention is the best and most effective way to work against extrinsic skin aging effects. The best prevention strategy against the harmful action of free radicals is a well regulated lifestyle

    (caloric restriction, body care and physical exercise for body), with low stress conditions and a balanced nutritional diet, includ- ing anti-oxidative rich food.

    Frequently researched antioxidants such as carotenoids, tocophenols and flavonoids, as well as vitamins (A, C, D and  E), essential omega-3-fatty acids, some proteins and lactobacilli have been referred as agents capable of promoting skin health and beauty.3,4 To find a proper balance, this review considers the beneficial “anti-aging” effects of increased reactive oxygen species (ROS) signaling recently.

    The appropriate generation of ROS (for instance after physi- cal exercise) has beneficial cell-protective and anti-aging effects. ROS activate via stimulation of STE-like 20 protein kinase 1 (MST1) and Jun N-terminus kinase (JNK) specific phosphor- ylations of forkhead box class O transcription factor (FoxO transcription factors), which thereafter translocate from the cyto- plasm into the nucleus and thereby induce the expression of anti- oxidative enzymes like superoxide dismutase, catalase and others. The expression and upregulation of the cell’s own intrinsic anti- oxidative enzyme systems finally do the “job” and protect the cell against accumulating and harmful cellular levels of ROS.5 Remarkably, upregulation of nuclear FoxO levels suppresses cell proliferation and induces apoptosis.

    The aim of this work is to review the existing literature and eventually to give an insight to the question whether diet actually influences the way our skin ages.

    Vitamins

    L-ascorbic acid (vitamin C). Vitamin C, also named L-ascorbic acid, is water soluble, photosensitive and is the most important antioxidant in the hydrophilic phase. Vitamin C is not naturally synthesized by the human body and therefore adequate dietary intake of vitamin C is required and essential for a healthy human diet.

    review

    The richest natural sources are fresh fruits and vegetables such as citrus fruits, blackcurrant, rose hip, guava, chili pepper or parsley. Stability of the vitamin C molecule depends on aggregate condition and formulation.

    L-ascorbic acid can be used orally and topically for skin benefits. Vitamin C is a cofactor for lysyl and prolyl hydroxy- lase, which stabilize the triple helical structure of collagen.6 It also plays a role in cholesterol synthesis, iron absorption and increases the bioavailability of selenium. The most commonly described cutaneous manifestations accompanying vitamin C deficiency are attributed to the impaired collagen synthesis. Enlargement and keratosis of hair follicles mainly of the upper arms and curled hairs, the so-called ‘corkscrew hairs’, are usu- ally described. The follicles become hemorrhagic with time and they sometimes mimic the palpable purpura of leucocytoclastic vasculitis.7

    Additionally, vitamin C deficiency is known for causing scurvy, a disease with some manifestations such as fragility, skin lesions in form of petechiae, gum bleeding, ease of developing bruises or slow wound healing.8

    Topically ascorbic acid is used in various cosmetic products, for example in lightening of skin dyspigmentation, anti-aging and sun protection formulations. The idea of sun protecting products is to have a combination product between a “passive” protection with a UV filter and an “active” protection with the antioxidant. UVB protection by vitamin C is frequently mentioned in the literature.6,9-11 However, the study by Wang et al. indicates that more work in formulation of cremes is needed, since there seem to be many products in which the desired effects are not measur- able.12 The use of vitamin C in cosmetic products is difficult as its reducing capacity occurs very fast and its degradation may occur under the presence of oxygen even before the topical application to the skin.13

    Nutricosmetic products with L-ascorbic acid work as free radi- cal scavengers and repair the membrane bound oxidized vitamin

    E.14 A long-term study observed the effects of a combination     of ascorbic acid and D-α-tocopherol (vitamin E) administered orally to human volunteers on UVB-induced epidermal damage. The treatment was well-tolerated and could be used prophylacti- cally against the hazardous effects of solar UV irradiation and skin cancer, according to the authors.9 Another paper describes an 8-week study, which compared topical and systemic antioxi- dant treatment. Topical and systemic treatment both seemed to be good photoprotectants.15

    There are many preparations of vitamin C- based products available on the market, but these are predominantly based on more stable esters and other derivatives of vitamin C which more readily penetrate the skin but are not necessarily converted to the only active vitamin C, L-ascorbic acid.16 These topical or oral products do not have the effects provided by L-ascorbic acid.

    Tocopherols (vitamin E). The vitamin E complex is a group of 8 compounds called tocopherols. Tocopherol is a fat-soluble membrane bound antioxidant and consequently a free-radical scavenger especially of highly reactive singlet oxygen. Tocopherol is like vitamin C a naturally occurring endogenous non-enzy- matic antioxidant.

    Vitamin C and vitamin E act synergistically. When UV-activated molecules oxidize cellular components, a chain reaction of lipid peroxidation in membranes rich in polyunsatu- rated fatty acids is induced. The antioxidant D-α-tocopherol is oxidized to the tocopheroxyl radical in this process and it is regen- erated by ascorbic acid to D-α-tocopherol.17,18 Beside ascorbic acid, glutathione and coenzyme Q10 can also recycle tocopherol. Higher amounts of tocopherol are available in vegetables, veg- etable oils like wheat germ oil, sunflower oil, safflower oil and seeds, corn, soy and some sorts of meat. The intake of natural vitamin E products helps against collagen cross linking and lipid

    peroxidation, which are both linked to aging of the  skin.

    With the process described above, D-α-tocopherol is involved in stabilizing the cell membrane by inhibiting oxidation of poly- unsaturated fatty acids, such as arachidonic acid of membrane phospholipids. Topical applied vitamin E is described to reduce erythema, sunburned cells, chronic UVB-induced skin damage and photocarcinogenesis in the majority of the published stud- ies.13,19 Vitamin E deficiency has been associated with a syndrome of edema with papular erythema or seborrhoiec changes, dryness and depigmentation in premature  infants.20

    There are many clinical studies, which have tested the effects of tocopherol. The data seem to be controversial, but high doses of oral vitamin E may affect the response to UVB in humans.21 Data of Ekanayake-Mudiyanselage and Thiele suggest that vita- min E levels are dependent on the density of sebaceous glands in the skin. In a 3-week study with daily oral supplementation of moderate doses of α-tocopherol significantly increased vitamin E levels measured in skin sites rich in sebaceous glands, such as the face. This should be considered when designing clinical vitamin E studies.22

    Oral combination treatments of vitamins C and E, partly with other photoprotective compounds, did increase the photoprotec- tive effects dramatically compared with monotherapies. Experts recommend that this synergetic interplay of several antioxidants should be taken into consideration in future research on cutane- ous photoprotection.23

    Carotenoids (vitamin A, β-carotene, astaxanthin, retinol).

    Carotenoids are vitamin A derivates like β-carotene, astaxanthin, lycopene and retinol, which are all highly effective antioxidants and have been documented to possess photoprotective properties. Findings of Scarmo et al. suggest that human skin, is relatively enriched in lycopene and β-carotene, compared with lutein and zeaxanthin, possibly reflecting a specific function of hydrocarbon carotenoids in human skin photoprotection.24

    β-carotene is the most prominent member of the group of carotenoids, natural colorants that can be found in the human diet.25 Compared with other carotenoids, the primary role of β-carotene is its provitamin-A activity. β-carotene can be cleaved by BCMO1  enzyme into 2 molecules of all-trans-retinal. There   is no difference between naturally occurring and chemically synthesized β-carotene. Furthermore,  β-carotene  can also act  as a lipid radical scavenger and as a singlet oxygen quencher, as demonstrated in vitro.26 Based on the distribution of BCMO1 in human tissues it seems that β-carotene metabolism takes place in a wide variety of organs, including the  skin.27

    Carrots, pumpkin, sweet potatoes, mangos and papaya are some examples of β-carotene containing fruits and vegetables.

    Upon dietary supplementation, β-carotene can be further enriched in skin, in which it is already a major carotenoid.28 β-carotene is an endogenous  photoprotector,  and  its  efficacy to prevent UV-induced erythema formation has been demon- strated in various studies.29,30 In healthy volunteers, a 12-week oral administration of β-carotene may result in a reduction of UV-induced erythema.31 Similar effects have been described in volunteers receiving a lycopene-rich diet.32

    The systemic photoprotecting effect of β-carotene depends both on dose and duration of treatment. In studies document- ing protection against UV-induced erythema, supplementation with carotenoids lasted for at least 7 weeks, with doses > 12 mg/d of carotenoids.31,33-35 With treatment periods of only 3–4 weeks, studies reported no protective effects.36 Furthermore, β-carotene supplementation can significantly reduce the rate of mitochondrial mutation in human dermal fibroblasts after UV irradiation.37

    Astaxanthin is found in microalgae, yeast, salmon,  trout,  krill, shrimp, crayfish and crustacea. Astaxanthin is biosynthe- sized by microalgae or phytoplankton, which are consumed by zooplankton or crustacea. They accumulate astaxanthin and, in turn are ingested by fish which then accrue astaxanthin in the food chain.38 Therefore, astaxanthin has considerable potential and promising applications in human health and nutrition39 and has been attributed an extraordinary potential for protecting the organism against a wide range of diseases (reviewed in refs. 40 and 41).

    The UV protective effects of algal extract containing 14% of astaxanthin compaired to synthetic astaxanthin have also been tested. The authors of this study reported that preincubation with synthetic astaxanthin or an algal extract could prevent UVA- induced alterations in cellular superoxide dismutase activity and decrease in cellular glutathione content.42

    In a study of Camera et al. the modulation of UVA-related injury by astaxanthin, canthaxanthin, and β-carotene for sys- temic photoprotection in human dermal fibroblasts has been com- pared.43 Astaxanthin showed a significant photoprotective effect and counteracted UVA-induced alterations to a great extent. The uptake of astaxanthin by fibroblasts was higher than that of can- thaxanthin and β-carotene, which lead to the assumption that the effect of astaxanthin toward photooxidative changes was stronger than that of the other substances. A recent study of Suganuma  et al. showed that astaxanthin could interfere with UVA-induced matrix-metalloproteinase-1 and skin fibroblast elastase/neutral endopeptidase expression.44 Both studies suggest that effects of UVA radiation, such as skin sagging or wrinkling can be pre- vented or at least minimized by topical or oral administration of astaxanthin.36,42,44

    Lycopene is a bright red carotene and carotenoid pigment and phytochemical found in tomatoes and other red fruits and vegeta- bles, such as red carrots, watermelons and papayas (but not straw- berries or cherries). Although lycopene is chemically a carotene, it has no vitamin A activity.

    β-carotene  and  lycopene  are  usually  the  dominating carot-

    enoids in human blood and tissues and are known to    modulate

    skin properties when ingested as supplements or as dietary prod- ucts. While they cannot be compared with sunscreen, there is evidence that they protect the skin against sunburn (solar ery- thema) by increasing the basal defense against UV light-mediated damage.45

    A study confirmed that the amounts of lycopene in plasma and skin are comparable to or even greater than those of β-carotene. When skin is exposed to UV light stress, more skin lycopene is destroyed compared with β-carotene, suggesting a role of lyco- pene in mitigating oxidative damage in tissues.46 Lycopene and tomato products are also mentioned for preventing cancer.47,48

    Retinol is important for the human body; however the body itself cannot synthesize it. Retinol, a fat-soluble unsaturated iso- prenoid like its two important metabolites retinaldehyde and reti- noic acid, is essential for growth, differentiation and maintenance of epithelial tissues and influences reproduction. In human skin two retinoid receptors are expressed, which can be activated by retinol and its metabolites.49

    Retinaldehyde, additionally being important for vision, is cre- ated by in vivo oxidation of retinol in a reversible process. The normal plasma concentration of vitamin A in humans is 0.35– 0.75  μg/ml.50,51

    Retinol must derive from diet. Natural retinol and retinol ester are contained in liver, milk, egg yolk, cheese and fatty fish etc. Naturally occurring and synthetic vitamin A (retinol) show simi- lar biological activities. Different retinol products, both for cos- metic (topical) and pharmaceutical (topical, systemic) use can be found on the market.

    In a review of topical methods to counteract skin wrinkling and irregular pigmentation of aging skin, Bayerl evaluates the effects of vitamin A acid derivatives, chemical peeling and bleach- ing agents. Also, the effects of UV protection by using sunscreens and topical antioxidants are reviewed.52 The topical retinoid treat- ments inhibit the UV-induced, MMP-mediated breakdown of collagen and protect against UV-induced decreases in procollagen expression.53-55

    Endogenous retinoids cannot be linked to the pathogenesis of common skin diseases like acne and psoriasis. Oral treatment with retinol or retinal derivatives has not been proposed as a possible anti-aging treatment. Humans require 0.8–1 mg or 2400–3000  IU vitamin A per day (1 IU = 0.3 μg).51

    Unfortunately the large CARET trial mentioned lung cancer- promoting effects of 25,000 IU retinyl palmitate combined with 30 mg β-carotene intake in smokers.56 Thus, the belief that chem- ical quenching of free radicals by natural compounds like reti- nyl palmitate and β-carotene exerts always beneficial effects has been challenged. Omenns data showed that an artificial systemic increase of antioxidants by dietary supplementation intended to modify UV erythema thresholds may have severe internal adverse effects which even may not only increase risk of cell aging but   of tumor promotion. However experts still recommend dietary intake of fruits and vegetable.

    Vitamin D. In humans vitamin D serves two functions, it acts as a prohormone and the human body can synthesize it itself through sun exposure. Skin is the major site for UV-B mediated vitamin D3, and 1,25-dihydroxy vitamin D3 synthesis. Smaller

    Table 1. recommended dietary allowances for vitamin  D

    Age

    Male

    Female          Pregnancy          Lactation

    0–12 months*

    400 iU

    (10 mcg)

    400 iU

    (10 mcg)

    1–13 years

    600 iU

    (15 mcg)

    600 iU

    (15 mcg)

    14–18 years

    600 iU

    (15 mcg)

    600 iU

    (15 mcg)

    600 iU

    (15 mcg)

    600 iU

    (15 mcg)

    19–50 years

    600 iU

    (15 mcg)

    600 iU

    (15 mcg)

    600 iU

    (15 mcg)

    600 iU

    (15 mcg)

    51–70 years

    600 iU

    (15 mcg)

    600 iU

    (15 mcg)

    >70 years

    800 iU

    (20 mcg)

    800 iU

    (20 mcg)

    *Ai, adequate intake; iU, international unit; mcg, microgram; 40 iU = 1 mcg.

    amounts of vitamin D2 and D3 come from the dietary intake of animal-based foods such as fatty fish or egg yolk. Some products like milk, cereals and margarine can be enriched with vitamin  D.

    Excess of vitamin D is stored in fat of the body and can result in toxic effects. This toxicity presents with nausea, vomiting, poor appetite, weakness, weight loss and constipation. Food-intake of vitamin D high enough to cause toxicity is very unlikely.

    The skin is one of the key tissues of the human body vitamin D endocrine system. It is important for a broad variety of inde- pendent physiological functions, which are reviewed in Reichrath et al.51 Besides its role in calcium homeostasis and bone integrity 1,25-dihydroxy vitamin D3 [1,25(OH)2D3] is also essential for numerous physiologic functions including immune response, release of inflammatory cytokines and regulation of growth and differentiation in normal and malignant tissues such as breast, lung and colon.51 1,25(OH)2D3 protects human skin cells from UV-induced cell death and apoptosis,57 inhibits the activation of stress-activated protein kinases,58 such as the c-Jun NH2-terminal kinase and p38, and suppresses IL-6 production. Several in vitro and in vivo studies have documented the protective effect of 1,25(OH)2D3 against UVB-induced skin damage and carcino- genesis.58,59 Furthermore, 1,25(OH)2D3 induces the expression  of antimicrobial peptide genes in human skin60 and plays a sig- nificant role in preventing opportunistic infections. With increas- ing age the capacity of the skin to produce vitamin D3 declines and consequently the protective effects of the vitamin. There are several factors contributing to this deficiency state among them behavioral factors, for example limited sun exposure or malnu- trition, which can be partially altered by behavior modification and various intrinsic factors like reduced synthetic capacity. In skin, the concentration of 7-dehydrocholesterol—a vitamin D3 precursor—showed an approximately 50% decline from age 20 y to age 80 y61 and the total amount of pre-vitamin D3 in the skin of young subjects was at least two times greater than when com- pared with that of the elderly subjects. Vitamin D and calcium supplementation is therefore of great importance in the elderly population.13

    Chang et al. also suggest an association between skin aging and levels of 25(OH)D3, another precursor of vitamin D. It may be possible that low 25(OH)D3 levels in women, who show less skin aging may reflect underlying genetic differences in vitamin D synthesis.62

    Many other studies that tested oral vitamin D treatment showed skin cancer prevention, which is linked to anti-aging effects.63,64

    In 2009, the American Academy of Dermatology and the Canadian Cancer Society recommended a  200  IU/day  dosis for children (0–14 y), 200 IU for the age population between 14–50 y, 400 IU for the 50–70 y and 600 IU for people over  their 71st year of age.65

    A higher dose of vitamin D 1000 IU/day (adults) and 400 IU/day (children 0–14 y) intake has been recommended for individuals with known risk factors for vitamin D insufficiency like dark skin individuals, elderly persons, photosensitive indi- viduals, people with limited sun exposure, obese individuals or those with fat  malabsorption.65

    The Food and Nutrition Board published a new recommenda- tion for dietary allowance levels and tolerable upper intake levels (ULs) for vitamin D intake in 2010. The recommended dietary allowance (Table 1) represents a daily intake that is sufficient to maintain bone health and normal calcium metabolism in healthy people.66

    Long-term intakes of vitamin D above the upper intake levels increase the risk of adverse health effects. Most reports suggest a toxicity threshold for vitamin D of 10,000 to 40,000 IU/day and serum 25(OH)D levels of 500–600 nmol/L (200–240 ng/mL).

    With daily intakes below 10,000 IU/day, toxicity symptoms are very unlikely. However, recent results from observational studies, national survey data and clinical trials have shown adverse health effects over time at much lower levels of vitamin D intakes and serum 25(OH)D. Since serum levels of approxi- mately 75–120 nmol/L or 30–48 ng/mL have been associated with increased all-cause mortality, greater risk of cancer at some sites like the pancreas, greater risk of cardiovascular events as well as more falls and fractures with elderly subjects, the Food and Nutrition Board advises that serum 25(OH)D levels above 125–150 nmol/L (50–60 ng/mL) should be avoided and cites research results that link vitamin D intakes of 5,000 IU/day  with a serum concentration at a maximum of 100–150 nmol/L (40–60 ng/mL).66

    Polyphenols

    Polyphenols have drawn the attention of the anti-aging research community over the last decade, mainly because of their antiox- idant properties, their great intake amount in our diet and the increasing studies showing their probable role in the prevention of various diseases associated with oxidative stress, such as cancer and cardiovascular and neurodegenerative diseases.67 Their total dietary intake could be as high as 1 g/d, which is much higher than that of all other classes of phytochemicals and known dietary antioxidants.68,69 They are mostly found in fruits and plant- derived beverages such as fruit juices, tea, coffee and red wine.

    Vegetables, cereals, chocolate and dry legumes are also sources for the total polyphenol intake.69 Several thousand molecules having a polyphenol structure have been identified in plants being gen- erally involved in defense against UV radiation or aggression by pathogens. Depending on the number of phenol rings and the way that these rings bind to one another, polyphenols can be divided into many different functional groups such as the phenolic acids, flavonoids, stilbenes, and lignans.67 Flavonoids are also further divided into flavones, flavonols, isoflavones, and flavanones, each with a slightly different chemical structure.6

    It has been reported that the polyphenolic content of foods can be easily affected or seriously reduced by methods of meal preparation and culinary traditions. For example, onions, which are a major source of phenolic acids and flavonoids, and tomatoes lose between 75% and 80% of their initial content when boiled over 15 min, 65% when cooked in a microwave oven and 30% when fried.70 In French fries or freeze-dried mashed potatoes no remaining phenolic acids were to be found.71

    Laboratory studies of different polyphenols such as, green tea polyphenols, grape seed proanthocyanidins, resveratrol, silyma- rin and genistein, conducted in animal models on UV-induced skin inflammation, oxidative stress and DNA damage, suggested that these polyphenols, combined with sunscreen protection, have the ability to protect the skin from the adverse effects of UV radiation, including the risk of skin cancers.72 The underlying mechanism of polyphenols actions has been a major discussion over the last decades. One of the most abundant theories is that the cells respond to polyphenols mainly through direct interac- tions with receptors or enzymes involved in signal transduction, which may result in modification of the redox status of the cell and may trigger a series of redox-dependent reactions.73,74 As antioxidants, polyphenols may improve cell survival; as prooxi- dants, they may induce apoptosis and prevent tumor growth.69,75 However, the biological effects of polyphenols may extend well beyond the modulation of oxidative stress.69

    Some interesting polyphenols, flavonoids and botanical anti- oxidants and their properties, which have drawn attention for their unique anti-aging effects are discussed next.

    Flavonoids. Phlorizin. Phlorizin belongs to the group of dihy- drochalcones, a type of flavonoids and it is naturally occurring  in some plants. It could be found in the bark of pear (Pyrus com- munis), apple, cherry and other fruit trees. It has been used as a pharmaceutical and tool for physiology research for over 150 y. However, its anti-aging effects have only been reported in the last years. Investigations of the effects of phlorizin on lifespan of the yeast Saccharomyces cerevisiae showed an improvement of the viability of the yeast, which was dose-dependent under oxidative stress.76 Further investigations on humans are needed.

    Many other botanical extracts, which are not discussed in  this review, have been described to have potent anti-oxidant properties. Among them silymarin,77 apigenin78 and  genistein79 have been demonstrated to have beneficial effects on skin aging parameters.

    Botanical anti-oxidants. The nutrient-sensitive kinase mam- malian target of rapamycin complex 1 (mTORC1) integrates nutrient signaling. This mTORC1  is the central hub    regulating

    protein and lipid synthesis, cell growth and cell proliferation and the process of autophagy and is thus intimately involved in cen- tral regulatory events associated with cell survival and cell aging. Intriguingly, all natural plant-derived polyphenols like EGCG, resveratrol, curcumin, genestin and others are  natural  inhibi- tors of mTORC1, recently described in this journal.80 Natural polyphenols exert their major metabolic activity as mTORC1 inhibitors, a fundament aspect relating calorie restriction and/or nutrient-derived mTORC1 attenuation to deceleration of aging. In fact, it has recently been demonstrated that mTORC1 inhibi- tion by rapamycin extended life span in mice.81 This antioxidants from naturals souce exhibit more crucial functions as “Botanical mTORC1 inhibitors” and attenuate mTORC1 signaling, a ben- eficial property which decelerates cell metabolism, energy expen- diture, mitochondrial activity and thus total ROS generation and oxidative stress load of the cells.

    Resveratrol (Stilbenes). Resveratrol is an antioxidant, natural polyphenol, abundant in the skin of grapes (but not in the flesh). It has been the subject of intense interest in recent years due to  a range of  unique  anti-aging  properties.  High  concentrations of natural resveratrol and resveratrol oligomeres are found in grape shoots from Vitis Vinifera. Resveratrol and its oligomeres, trans-piceatannol, the dimers epsilon-viniferin, ampelopsin, iso- epsilon-viniferin, the trimers miyabenol C and the tetramers hopeaphenol, R-viniferin and R2-viniferin belong to the sub- group of stilbenes. Resveratrol works both as a chelating agent and as a radical scavenger and in addition it takes part in inflam- mation  by  inhibiting  the  production  of  IL-8  by LPS-induced

    MAPK phosphorylation and a block of NFkB activation.82 In

    2002 Bhat et al. reported that resveratrol possesses cancer chemo- preventive activities.83 Cardiovascular benefits via increased nitric oxide production, downregulation of vasoactive peptides, lowered levels of oxidized low-density lipoprotein, and cyclooxygenase inhibition; possible benefits on Alzheimer disease by breakdown of β-amyloid and direct effects on neural tissues; phytohormonal actions; antimicrobial effects; and sirtuin activation, which is believed to be involved in the caloric restriction-longevity effect have also been reported.84 As far as skin is concerned, resveratrol has been recently shown to possess a protective action in vitro against cell death after exposure of HaCaT cells to the nitric oxide free radical donor sodium nitroprusside.85 Furthermore, Giardina et al. reported in 2010 that in experiments in vitro with skin fibroblasts treated with resveratrol there was a dose-related increase in the rate of cell proliferation and in inhibition of col- lagenase activity.86 Steinberg showed that resveratrol oligomers hopeaphenol, epsilon-viniferin, R2-viniferin, ampelopsin inhibit the growth number of human tumor cell lines significantly stron- ger than resveratrol itself.87,88

    Curcumin. Curcumin is the principal curcuminoid of the pop- ular Indian spice turmeric, which is a member of the ginger family (Zingiberaceae) and is frequently found in rice dishes to add yel- low color to the otherwise white rice. Curcumin has been shown to protect against the deleterious effects of injury by attenuating oxi- dative stress and suppressing inflammation (reviewed in ref. 89). In human fibroblasts curcumin induced cellular stress responses through  phosphatidylinositol  3-kinase/Akt pathway  and redox

    signaling, thus providing evidence that curcumin-induced hor- metic stimulation of cellular antioxidant defenses can be a use- ful approach toward anti-aging intervention.90 Oral ingestion in rodents has produced correction of cystic fibrosis defects and inhi- bition of tumor proliferation, but human trials are  lacking.6,91,92

    Green tea polyphenols. Green tea polyphenols (GTPs) derivat- ing from the leaves of the Camellia sinensis have been postulated to protect human skin from the cutaneous signs of photoageing. In animal models, UV-induced cutaneous edema and cyclooxy- genase activity could be significantly inhibited by feeding the animals with GTPs.93 However, in a study in 2005, although participants treated with a combination regimen of topical and oral green tea showed histologic improvement in elastic tissue content, clinically significant changes could not be detected.94 Many laboratories have reported that topical treatment or oral consumption of green tea polyphenols inhibits chemical carcino- gen- or UV radiation-induced skin tumorigenesis in different animal models. Studies have shown that green tea extract also possesses anti-inflammatory activity. These anti-inflammatory and anti-carcinogenic properties of green tea are due to their polyphenolic constituents present therein. The major and most chemopreventive constituent in green tea responsible for these biochemical or pharmacological effects is (-)-epigallocatechin- 3-gallate (EGCG).95 EGCG can directly inhibit the expression of

    metalloproteinases such as MMP-2, MMP-9 and MMP-12,96 and is a potent inhibitor of leucocyte elastase,97 which is instrumental in tumor invasion and metastasis.

    Topical application of green tea extract containing GTPs on C3H mice reduced UVB- induced inflammation.98 The research- ers also found protection against UV-induced edema, erythema, and antioxidant depletion in the epidermis. This work further investigated the effects of GTPs after application to the back of humans 30 min before UV irradiation. A decrease of myeloper- oxidase activity and infiltration of leukocytes compared with the untreated skin was documented.99

    Ubiquinol (Coenzyme Q10)

    Coenzyme Q10 (CoQ10) is a fat-soluble, endogenous (synthe- sized by the body), vitamin-like substance that is mainly stored in the fat tissues of our body. It is present in most eukaryotic cells, primarily in the mitochondria and plays an important role as a component of the electron transport chain in the aerobic cellular respiration, generating energy. Ubiquinol is also a well- known powerful antioxidant compound. In the skin, CoQ10 is mainly to be found in the epidermis where it acts in combination with other enzymic and non-enzymic substances as the initial barrier to oxidant assault.100 Primary dietary sources of CoQ10 include oily fish (such as salmon and tuna), organ meats (such   as liver), and whole grains. The amount of CoQ10 needed in human organism can be gained through a balanced diet, how- ever in the market CoQ10 is available in several forms as a supple- ment, including soft gel capsules, oral spray, hard shell capsules, and tablets. As a fat-soluble substance it is better absorbed when taken with fat rich meals. CoQ10 is also added to various cos- metics. It has been shown on rats that a CoQ   supplementation

    elevates CoQ homologs in tissues and their mitochondria, thus causing a selective decrease in protein oxidative damage, and an increase in antioxidative potential.101 Furthermore, in a human study where 50 mg each of vitamin E, coenzyme Q10, and sele- nium were administered combined with the use of topical bio- cosmetics, an increase in stratum corneum CoQ10 was noted after 15 and 30 d of ingestion.102 In cases of primary CoQ10  deficiency in vitro experiments have shown that they should be treated with CoQ10 supplementation and that complementary administration of antioxidants with  high bioavailability  should be considered if oxidative stress is present.103 On the other hand, in experiments contacted on mice the supplemental intake of CoQ10 had no effect on the main antioxidant defense or pro- oxidant generation in most tissues, and had no impact on the life span of mice.104

    Pre- and Probiotics

    The term probiotic is defined as “living microorganisms, which, when consumed in adequate amounts, confer a health effect on the host.”105,106

    The most commonly used probiotics in humans and animals are enterococci, lactobacilli and bifidobacteria, which are natural residents of the intestinal tract.

    A prebiotic is a non-viable food component that confers a health benefit on the host associated with modulation of the microbiota.107 Oligofructose and other oligosaccharides are prebi- otic which have a significant effect on the population of luminal flora, in particular, stimulating bifidobacterial   populations.

    Currently, finding alternatives to antibiotics for skin treat- ment is receiving a lot of interest in research. It has been found that, similarly to the gut microflora, the skin’s microbiota plays a beneficial role. Thus, the possibility to modulate the microbiota more selectively is highly interesting.

    UV exposure is known to negatively  affect  immune  sys- tem functions.108 Clinical studies that used probiotic bacteria (Lactobacillus johnsonii NCC 533) to modulate the cutaneous immune homeostasis altered by solar-simulated UV exposure in humans suggest that certain probiotics can help preserve the skin homeostasis by modulating the skin immune  system.109,110

    According to Schouten et al., a prebiotic diet caused reduced acute allergic skin response in recipient mice.111

    Essential Fatty Acids (Vitamin F)

    Essential fatty acids (EFAs) are long-chain polyunsaturated fatty acids derived from linolenic, linoleic and oleic acids. They can- not be produced in the human body and they have to be con- sumed through our daily dietary intake. EFAs have also been known as vitamin F. Arachidonic acid is a semi-EFA, as it can be synthesized in the body from linoleic acid. The two families of EFAs are ω-3, derived from linolenic acid, and ω-6, derived from linoleic acid, with the number indicating the position of  the first double bond continuing from the terminal methyl group on the molecule.6,112 They are present in multiple food sources such as fish and shellfish, flaxseed, hemp oil, soya oil, canola oil,

    chia seeds, pumpkin seeds, sunflower seeds, leafy vegetables, wal- nuts, sesame seeds, avocados, salmon and albacore tuna. EFAs are essential for the synthesis of tissue lipids, play an important role in the regulation of cholesterol levels and are precursors of prostaglandins.113

    The association between nutrient intakes and skin aging has been examined in 2008 in 4025 women (40–74 y), using data from the first National Health and Nutrition Examination Survey. Skin-aging appearance was defined as having a wrinkled appearance, senile dryness, and skin atrophy. Higher linoleic acid intakes were associated with a lower likelihood of senile dryness and skin atrophy.114 In a study where the effect of fish oil on UV (UV) B-induced prostaglandin metabolism was examined, 13 patients with polymorphic light eruption received dietary supple- ments of fish oil rich in omega-3 polyunsaturated fatty acids for 3 mo. The authors managed to show a reduction in UV-induced inflammation, possibly due to lowered prostaglandin-E2 lev- els.115 Furthermore, oral administration of an antioxidant mix- ture containing vitamin C, vitamin E, pycnogenol and evening primrose oil significantly inhibited wrinkle formation caused by chronic UVB irradiation through significant inhibition of UVB- induced matrix metalloproteinase (MMP) activity accompanied by enhancement of collagen synthesis on hairless mouse skin.116

    EFAs can also be found as artificial supplements in the mar- ket. Fish oil supplements are usually made from mackerel, her- ring, tuna, halibut, salmon, cod liver, whale blubber, or seal blubber, are rich in omega-3 fatty acids and often contain small amounts of vitamin E. They might be also combined with cal- cium, iron, or vitamins A, B1, B2, B3, C or  D.

    Caloric Restriction

    It is widely accepted that caloric restriction (CR), without malnutrition, delays the onset of aging and extends lifespan in

    diverse animal models including yeast, worms, flies, and labo- ratory rodents.117 Although the underlying mechanisms remain still unknown, some explanations such as alterations of hor- mone metabolism, hormone-related cellular signaling, oxidation status, DNA repair, apoptosis, and oncogene expression, have been postulated.118,119 In a histological  study  on  Fischer  344  rats undergoing dietary CR, the histomorphological changes resulting from intrinsic aging were delayed or prevented by CR. Namely, a trend toward increased values for collagen and elastic fibers, fibroblasts, and capillaries and a prevention of age-related increase in the depth of the epidermis, dermis, and fat layer was observed in skin samples from CR rats.120 Furthermore, in skin tissues of mice with CR weight control a palette of genes showed a differential expression when compared with mice receiving normal diet. The authors concluded that dietary CR showed profound inhibitory impact on the expression of genes relevant to cancer risks.121 Studies evaluating CR in nonhuman primates and its effects on human health, and on the metabolic param- eters are ongoing.

    Conclusions

    To conclude, nutrition and skin aging still remains a controver- sial and conflicting subject. A promising strategy for enhancing skin protection from oxidative stress is to support the endoge- nous antioxidant system, with antioxidants containing products that are normally present in the skin.11 However, this should be not confused with a permanent intake of non-physiological high dosages of isolated antioxidants. Fruit and vegetables consump- tion may represent the most healthy and safe method in order to maintain a balanced diet and youthful appearing   skin.

    Disclosure of Potential Conflicts of Interest

    No potential conflicts of interest were disclosed.

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  • Astaxanthin: Sources, Extraction, Stability, Biological Activities and Its Commercial Applications—A Review

    Ranga Rao Ambati 1,*, Siew Moi Phang 1, Sarada Ravi 2 and Ravishankar Gokare Aswathanarayana 3

    1    Institute of Ocean and Earth Sciences, University of Malaya, Kuala Lumpur 50603, Malaysia; E-Mail: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.

    2    Plant Cell Biotechnology Department, Central Food Technological Research Institute, (Constituent

    Laboratory of Council of Scientific & Industrial Research), Mysore-570020, Karnataka, India; E-Mail: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.

    3    C. D. Sagar Centre for Life Sciences, Dayananda Sagar Institutions, Kumaraswamy Layout,

    Bangalore-560078, Karnataka, India; E-Mail: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.

     

    * Author to whom correspondence should be addressed; E-Mail: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.; Tel.: +603-79674610; Fax: +603-79676994.

     

    Received: 10 October 2013; in revised form: 10 December 2013 / Accepted: 11 December 2013 /

    Published: 7 January 2014

     
       

     

     

    Abstract: There is currently much interest in biological active compounds derived from natural resources, especially compounds that can efficiently act on molecular targets, which are involved in various diseases. Astaxanthin (3,3′-dihydroxy-β, β′-carotene-4,4′-dione) is a xanthophyll carotenoid, contained in Haematococcus pluvialis, Chlorella zofingiensis, Chlorococcum, and Phaffia rhodozyma. It accumulates up to 3.8% on the dry weight basis in H. pluvialis. Our recent published data on astaxanthin extraction, analysis, stability studies, and its biological activities results were added to this review paper. Based on our results and current literature, astaxanthin showed potential biological activity in in vitro  and in vivo models. These studies emphasize the influence of astaxanthin and its beneficial effects on the metabolism in animals and humans. Bioavailability of astaxanthin in animals was enhanced after feeding Haematococcus biomass as a source of astaxanthin. Astaxanthin, used as a nutritional supplement, antioxidant and anticancer agent, prevents diabetes, cardiovascular diseases, and neurodegenerative disorders, and also stimulates immunization. Astaxanthin products are used for commercial applications in the dosage forms as tablets, capsules, syrups, oils, soft gels, creams, biomass and granulated powders. Astaxanthin patent applications are available in food, feed and nutraceutical    applications.

     

     

    The current review provides up-to-date information on astaxanthin sources, extraction, analysis, stability, biological activities, health benefits and special attention paid to its commercial applications.

     

    Keywords:   astaxanthin;    sources;    stability;    biological    activities;    health    benefits; applications

     
       

     

     

     

    1.  Introduction

     

    Astaxanthin is a xanthophyll carotenoid which is found in various microorganisms and marine animals [1]. It is a red fat-soluble pigment which does not have pro-Vitamin A activity in the human body, although some of the studies reported that astaxanthin has more potent biological activity than other carotenoids. The United States Food and Drug Administration (USFDA) has approved the use of astaxanthin as food colorant in animal and fish feed [2]. The European Commission considers natural astaxanthin as a food dye [3]. Haematococcus pluvialis is a green microalga, which accumulates high astaxanthin content under stress conditions such as high salinity, nitrogen deficiency, high temperature and light [4–6]. Astaxanthin produced from H. pluvialis is a main source for human consumption [7].  It is used as a source of pigment in the feed for salmon, trout and shrimp [1,3]. For dietary supplement in humans and animals, astaxanthin is obtained from seafood or extracted from H. pluvialis [8]. The consumption of astaxanthin can prevent or  reduce  risk  of  various  disorders  in  humans  and  animals [7,8]. The effects of astaxanthin on human health nutrition have been published by various authors [7–13]. In our previous reviews, we included recent findings on the potential effects of astaxanthin and its esters on biological activities [14–18]. The use of astaxanthin as a nutritional supplement has been rapidly growing in foods, feeds, nutraceuticals and pharmaceuticals. This present review paper provides information on astaxanthin sources, extraction methods, storage stability, biological activities, and health benefits for the prevention of various  diseases  and  use  in  commercial applications.

     

    2.  Source of Astaxanthin

     

    The natural sources of astaxanthin are algae, yeast, salmon, trout, krill, shrimp and crayfish. Astaxanthin from various microorganism sources are presented in Table 1. The commercial  astaxanthin is mainly from Phaffia yeast, Haematococcus and through chemical synthesis. Haematococcus pluvialis is one of the best sources of natural astaxanthin [17–20]. Astaxanthin content in wild and farmed salmonids are shown in Figure 1. Among the wild salmonids, the maximum astaxanthin content in wild Oncorhynchus species was reported in the range of 26–38 mg/kg flesh in sockeye salmon whereas low astaxanthin content was reported in chum [20]. Astaxanthin content in farmed Atlantic salmon was reported as 6–8 mg/kg flesh. Astaxanthin is available in the European      (6 mg/kg flesh) and Japanese market (25 mg/kg flesh) from large trout. Shrimp, crab and salmon can serve as dietary sources of astaxanthin [20]. Wild caught salmon is a good source of astaxanthin. In

     

     

    order to get 3.6 mg of astaxanthin one can eat 165 grams of salmon per day. Astaxanthin supplement at

    3.6 mg per day can be beneficial to health as reported by Iwamoto et al. [21].

     

    Table 1. Microorganism sources of astaxanthin.

     

    Sources

    Astaxanthin (%) on the Dry Weight Basis

    References

    Chlorophyceae

     

     

    Haematococcus pluvialis

    3.8

    [17,18]

    Haematococcus pluvialis (K-0084)

    3.8

    [22]

    Haematococcus pluvialis (Local isolation)

    3.6

    [23]

    Haematococcus pluvialis (AQSE002)

    3.4

    [24]

    Haematococcus pluvialis (K-0084)

    2.7

    [25]

    Chlorococcum

    0.2

    [26,27]

    Chlorella zofingiensis

    0.001

    [28]

    Neochloris wimmeri

    0.6

    [29]

    Ulvophyceae

     

     

    Enteromorpha intestinalis

    0.02

    [30]

    Ulva lactuca

    0.01

    [30]

    Florideophyceae

     

     

    Catenella repens

    0.02

    [30]

    Alphaproteobacteria

     

     

    Agrobacterium aurantiacum

    0.01

    [31]

    Paracoccus carotinifaciens (NITE SD 00017)

    2.2

    [32]

    Tremellomycetes

     

     

    Xanthophyllomyces dendrorhous (JH)

    0.5

    [33]

    Xanthophyllomyces dendrorhous (VKPM Y2476)

    0.5

    [34]

    Labyrinthulomycetes

     

     

    Thraustochytrium sp. CHN-3 (FERM P-18556)

    0.2

    [35]

    Malacostraca

     

     

    Pandalus borealis

    0.12

    [20]

    Pandalus clarkia

    0.015

    [36]

     

    Figure 1. Astaxanthin levels (mg/kg flesh) of wild and farmed (*) salmonids [20].

     
       

     

     

     

    3.  Structure of Astaxanthin

     

    Astaxanthin is a member of the xanthophylls, because it contains not only carbon and hydrogen but also oxygen atoms (Figure 2). Astaxanthin consists of two terminal rings joined by a polyene chain. This molecule has two asymmetric carbons located at the 3, 3′ positions of the β-ionone ring with hydroxyl group (-OH) on either end of the molecule. In case one, hydroxyl group reacts with a fatty acid then it forms mono-ester, whereas when both hydroxyl groups are reacted with fatty acids the result is termed a di-ester. Astaxanthin exists in stereoisomers, geometric isomers, free and esterified forms [1]. All of these forms are found in natural sources. The stereoisomers (3S, 3S) and (3R 3′R) are the most abundant in nature. Haematococcus biosynthesizes the (3S, 3′S)-isomer whereas yeast Xanthophyllomyces dendrorhous produces (3R, 3′R)-isomer [10]. Synthetic astaxanthin comprises isomers of (3S, 3′S) (3R, 3′S) and (3R, 3′R). The primary stereoisomer of astaxanthin found in the Antarctic krill Euphausia superba is 3R, 3′R which contains mainly esterified form, whereas in wild Atlantic salmon it is 3S, 3′S which occurs as the free form [37]. The relative percentage of astaxanthin and its esters in krill, copepod, shrimp and shell is shown in Figure 3. Astaxanthin has the molecular formula C40H52O4. Its molar mass is 596.84 g/mol.

    Figure 2. Planner structure of astaxanthin.

     
       

     

     

    Figure 3. Astaxanthin and its esters from various sources [19,20].

     
       

     

     

     

    4.  Extraction and Analysis of Astaxanthin

     

    Astaxanthin is a lipophilic compound and can be dissolved in solvents and oils. Solvents, acids, edible oils, microwave assisted and enzymatic methods are used for astaxanthin extraction.  Astaxanthin is accumulated in encysted cells of Haematococcus. Astaxanthin in Haematococcus was extracted with different acid treatments, hydrochloric acid giving up  to  80%  recovery  of  the  pigment [38]. When encysted cells were treated with 40% acetone at 80 °C for 2 min followed by kitalase, cellulose, abalone and acetone powder, 70% recovery of astaxanthin was obtained [39]. High astaxanthin yield was observed with treatment of hydrochloric acid at various temperatures for 15 and 30 min using sonication [40]. In another study, vegetable oils (soyabean, corn, olive and grape seed) were used to extract astaxanthin from Haematococcus. The culture was mixed with oils, and the astaxanthin inside the cell was extracted into the oils, with the highest  recovery of 93% with olive     oil [41]. Astaxanthin (1.3 mg/g) was extracted from Phaffia rhodozyma under acid conditions [42]. Microwave assisted extraction at 75 °C for 5 min resulted in 75% of astaxanthin; however, astaxanthin content was high in acetone extract [43,44]. Astaxanthin yield from Haematococcus was 80%–90% using supercritical fluid extraction with ethanol and sunflower oil as co-solvent [45–47]. Astaxanthin was extracted repeatedly with solvents, pooled and evaporated by rotary evaporator, then re-dissolved in solvent and absorbance of extract was measured at 476–480 nm to  estimate  the  astaxanthin  content [17]. Further the extract can be analyzed for quantification of astaxanthin using high pressure liquid chromatography and identified by mass spectra [18].

     

    5.  Storage and Stability of Astaxanthin

     

    Astaxanthin stability was assessed in various carriers and storage conditions. Astaxanthin derived from Haematococcus and its stability in various edible oils was determined [48]. Astaxanthin was stable at 70–90 °C in ricebran, gingelly and palm oils with 84%–90% of retention of astaxanthin content which can be used in food, pharmaceutical and nutraceutical applications, whereas astaxanthin content was reduced at 120 and 150 °C [48]. Astaxanthin nanodispersions’ stability was evaluated in skimmed milk, orange juice and deionized water was used as a control [49]. It was found that degradation of astaxanthin was significantly higher in skimmed milk than orange juice. In another study, stability of astaxanthin biomass was examined after drying and storage at various conditions for nine weeks [50]. The results showed that degradation of astaxanthin was as low as 10% in biomass dried at 180/110 °C and stored at −21 °C under nitrogen after nine weeks of storage. The stability of astaxanthin from Phaffia rhodozyma was studied and it was found that stability was high at pH 4.0 and at a lower temperature [51]. The storage stability of astaxanthin was enhanced at 4 °C and 25 °C in a complex mixture of hydroxyproply-β-cyclodextrin and water [52]. Astaxanthin stability was investigated using microencapsulation with chitosan, polymeric nanospheres, emulsions and β-cyclodextrin as reported by various authors [53–56].

     

    6.  Biochemistry of Astaxanthin

     

    Astaxanthin contains conjugated double bonds, hydroxyl and keto groups. It has both lipophilic and hydrophilic properties [1]. The red color is due to the conjugated double bonds at the center of the

     

     

    compound. This type of conjugated double bond acts as a strong antioxidant by donating the electrons and reacting with free radicals to convert them to be more stable product and terminate free radical chain reaction in a wide variety of living organisms [8]. Astaxanthin showed better biological activity than other antioxidants [11], because it could link with cell  membrane  from  inside  to  outside  (Figure 4).

     

    Figure 4. Superior position of astaxanthin in the cell membrane [12].

     
       

     

     

    7.  Bioavailability and Pharmacokinetics of Astaxanthin

     

    • Bioavailability

     

    Dietary oils may enhance the absorption of astaxanthin. Astaxanthin with combination of fish oil promoted hypolipidemic/hypocholesterolemic effects in plasma and its increased phagocytic activity  of activated neutrophils when compared with astaxanthin and fish oil alone [57]. Astaxanthin was superior to fish oil in particular by improving immune response and lowering the risk of vascular and infectious diseases. The proliferation activity of T- and B-lymphocytes was diminished followed by lower levels of O2, H2O2 and NO production, increased antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase (GPx), and calcium release in cytosol after administration of astaxanthin with fish oil [58]. Bioavailability and antioxidant properties of astaxanthin were enhanced in rat plasma and liver tissues after administration of Haematococcus biomass dispersed in olive        oil [14,15,17].

    Astaxanthin is a fat soluble compound, with increased absorption when consumed with dietary oils. Astaxanthin was shown to significantly influence immune function in several in vitro and in vivo assays [14,15,17]. Lipophilic compounds such as astaxanthin are usually transformed metabolically before they are excreted, and metabolites of astaxanthin have been detected in various rat tissues [59]. Astaxanthin bioavailability in human plasma was confirmed with single dosage of 100 mg [60]. Its

     

     

    accumulation in humans was found after administration of Haematococcus biomass as source of astaxanthin [61]. Astaxanthin bioavailability in humans was enhanced by lipid based formulations; high amounts of carotenes solubilized into the oil phase of the food matrix can lead to greater bioavailability [62]. A recent study reported that astaxanthin accumulation in rat plasma and liver was observed after feeding of Haematococcus biomass as source of astaxanthin [14,15,17].

     

    • Pharmacokinetics

     

    Carotenoids are absorbed into the body like lipids and transported via the lymphatic system into the liver. The absorption of carotenoids is dependent on the accompanying dietary components. A high cholesterol diet may increase carotenoid absorption while a low fat diet reduces its absorption. Astaxanthin mixes with bile acid after ingestion and make micelles in the intestinum tenue. The micelles with astaxanthin are partially absorbed by intestinal mucosal cells. Intestinal mucosal cells incorporate astaxanthin into chylomicra. Chylomicra with astaxanthin are digested by lipoprotein  lipase after releasing into the lymph within the systemic circulation, and chylomicron remnants are rapidly removed by the liver and other tissues. Astaxanthin is assimilated with lipoproteins and transported into the tissues [62]. Of several naturally occurring carotenoids, astaxanthin is considered one of the best carotenoids being able to protect cells, lipids and membrane lipoproteins against oxidative damage.

     

    8.  Biological Activities of Astaxanthin and Its Health Benefits

     

    • Antioxidant Effects

     

    An antioxidant is a molecule which can inhibit oxidation. Oxidative damage is initiated by free radicals and reactive oxygen species (ROS). These molecules have very high reactivity and are produced by normal aerobic metabolism in organisms. Excess oxidative molecules may react with proteins, lipids and DNA through chain reaction, to cause protein and lipid oxidation and DNA  damage which are associated with various disorders. This type of oxidative molecules can be inhibited by endogenous and exogenous antioxidants such as carotenoids. Carotenoids contain polyene chain, long conjugated double bonds, which carry out antioxidant activities by quenching singlet oxygen and scavenging radicals to terminate chain reactions. The biological benefits of carotenoids may be due to their antioxidant properties attributed to their physical and chemical interactions with cell membranes. Astaxanthin had higher antioxidant activity when compared to various carotenoids such as lutein, lycopene, α-carotene and β-carotene reported by Naguib et al. [63]. The antioxidant enzymes catalase, superoxide dismutase, peroxidase and thiobarbituric acid reactive substances (TBARS) were high in  rat plasma and liver after feeding Haematococcus biomass as source of astaxanthin [17]. Astaxanthin in H. pluvialis offered the best protection from free radicals in rats followed by β-carotene and       lutein [15,17]. Astaxanthin contains a unique molecular structure in the presence of hydroxyl and keto moieties on each ionone ring, which are responsible for the high antioxidant properties [10,64]. Antioxidant activity of astaxanthin was 10 times more than zeaxanthin, lutein, canthaxanthin, β-carotene and 100 times higher than α-tocopherol [65]. The oxo functional group in carotenoids has higher antioxidant activity without pro-oxidative contribution [66]. The polyene chain in astaxanthin

     

     

    traps radicals in the cell membrane, while the terminal ring of astaxanthin could scavenge radicals at the outer and inner parts of cell membrane (Figure 4). Antioxidant enzyme activities were evaluated in the serum after astaxanthin was supplemented in the diet of rabbits, showing enhanced activity of superoxide dismutase and thioredoxin reductase whereas paraoxonase was inhibited in the oxidative-induced rabbits [67]. Antioxidant enzyme levels were increased when astaxanthin fed to ethanol-induced gastric ulcer rats [68].

     

    • Anti-Lipid Peroxidation Activity

     

    Astaxanthin has a unique molecular structure which enables it to stay both in and outside the cell membrane. It gives better protection than β-carotene and Vitamin C which can be positioned inside the lipid bilayer. It serves as a safeguard against oxidative damage by various mechanisms, like quenching of singlet oxygen; scavenging of radicals to prevent chain reactions; preservation of membrane structure by inhibiting lipid peroxidation; enhancement of immune system function and regulation of gene expression. Astaxanthin and its esters showed 80% anti-lipid peroxidation activity in ethanol induced gastric ulcer rats and skin cancer rats [14,68]. Astaxanthin inhibited lipid peroxidation in biological samples reported by various authors [14,15,17,18,68,69].

     

    • Anti-Inflammation

     

    Astaxanthin is a potent antioxidant to terminate the induction of inflammation in biological  systems. Astaxanthin acts against inflammation. Algal cell extracts of Haematococcus and Chlorococcum significantly reduced bacterial load and gastric inflammation in H. pylori-infected   mice [16,70,71]. Park et al. [72] reported astaxanthin reduced the DNA oxidative damage biomarker inflammation, thus enhancing immune response in young healthy adult female human  subjects.  Haines et al. [73] reported lowered bronchoalveolar lavage fluid inflammatory cell numbers, and enhanced cAMP, cGMP levels in lung tissues after feeding astaxanthin with Ginkgo biloba extract and Vitamin C. Another study showed astaxanthin esters and total carotenoids from Haematococcus exerted a dose-dependent gastroprotective effect on acute, gastric lesions in ethanol-induced gastric ulcers in rats. This may be due to inhibition of H1, K1 ATPase, upregulation of mucin content and an increase in antioxidant activities [68]. Astaxanthin showed protective effect on high glucose induced oxidative stress, inflammation and apoptosis in proximal tubular epithelial cells. Astaxanthin is a promising molecule for the treatment of ocular inflammation in eyes as reported by the Japanese researchers [74,75]. Astaxanthin can prevent skin thickening and reduce collagen reduction against UV induced skin damage [14,76,77].

     

    • Anti-Diabetic Activity

     

    Generally, oxidative stress levels are very high in diabetes mellitus patients. It is induced by hyperglycemia, due to the dysfunction of pancreatic β-cells and tissue damage in patients. Astaxanthin could reduce the oxidative stress caused by hyperglycemia in pancreatic β-cells and also improve glucose and serum insulin levels [78]. Astaxanthin can protect pancreatic β-cells against glucose toxicity.  It  was  also  shown  to  be  a  good  immunological  agent  in  the  recovery  of    lymphocyte

     

     

    dysfunctions associated with diabetic rats [79]. In another study, ameliorate oxidative stress in streptozotocin-diabetes rats were inhibited by the combination of astaxanthin with α-tocopherol [80]. It is also inhibited glycation and glycated protein induced cytotoxicity in human umbilical vein endothelial cells by preventing lipid/protein oxidation [81]. Improved insulin sensitivity in both spontaneously hypertensive corpulent rats and mice on high fat plus high fructose diets was observed after feeding with astaxanthin [82–84]. The urinary albumin level in astaxanthin treated diabetic mice was significantly lower than the control group [78]. Some of the studies demonstrated that astaxanthin prevents diabetic nephropathy by reduction of the oxidative stress and renal cell damage [85–87].

     

    • Cardiovascular Disease Prevention

     

    Astaxanthin is a potent antioxidant with anti-inflammatory activity and its effect examined in both experimental animals and human subjects. Oxidative stress and inflammation are pathophysiological features of atherosclerotic cardiovascular disease. Astaxanthin is a potential therapeutic agent against atherosclerotic cardiovascular disease [88]. The efficacy of disodium disuccinate astaxanthin (DDA) in protecting mycocardium using mycocardial ischemia reperfusion model in animals was evaluated. Myocardial infarct size was reduced in Sprague Dawley rats, and improved in myocardial salvage in rabbits after four days of pre-treatment with DDA at 25, 50 and 75 mg/kg body weight [89,90]. Astaxanthin was found in rat mycocardial tissues after pretreatment with DDA at dosage of 150 and

    500 mg/kg/day for seven days [91]. Astaxanthin effects on blood pressure in spontaneously hypertensive rats (SHR), normotensive Wistar Kyoto rats (NWKR) and stroke prone spontaneously hypertensive rats (SPSHR) were reported [92]. Astaxanthin was found in the plasma, heart, liver, platelets, and increased basal arterial blood flow in mice fed with astaxanthin derivative [93]. Human umbilical vien endothelial cells and platelets treated with the astaxanthin showed increased nitric oxide levels and decrease in peroxynitrite levels [93]. Mice fed 0.08% astaxanthin had higher heart mitochondrial membrane potential and contractility index compared to the control group [94]. Astaxanthin effects on paraoxonase, thioredoxin reductase activities, oxidative stress parameters and lipid profile in hypercholesterolemic rabbits were evaluated. Astaxanthin prevented the activities of those enzymes from hypercholesterolemia induced protein oxidation at the dosages of  100 mg and  500 mg/100 g [67].

     

    • Anticancer Activity

     

    The specific antioxidant dose may be helpful for the early detection of various degenerative disorders. Reactive oxygen species such as superoxide, hydrogen peroxide and hydroxyl radical are generated in normal aerobic metabolism. Singlet oxygen is generated by photochemical events  whereas peroxyl radicals are produced by lipid peroxidation. These oxidants contribute to aging and degenerative diseases such as cancer and atherosclerosis through oxidation  of DNA, proteins  and lipids [95]. Antioxidant compounds decrease mutagenesis and carcinogenesis by inhibiting oxidative damage to cells. Cell–cell communication through gap junctions is lacking in human tumors and its restoration tends to decrease tumor cell proliferation. Gap junctional communication occurs due to an increase in the connexin-43 protein via upregulation of the connexin-43 gene. Gap junctional communication  was  improved  in  between  the  cells  by  natural  carotenoids  and  retinoids      [96].

     

     

    Canthaxanthin and astaxanthin derivatives enhanced gap junctional communication between mouse embryo fibroblasts [97–99]. Increased connexin-43 expression in murine fibroblast cells by β-carotene was reported [100,101]. Astaxanthin showed significant antitumor activity when compared to other carotenoids like canthaxanthin and β-carotene [102,103]. It also inhibited the growth of fibrosarcoma, breast, and prostate cancer cells and embryonic fibroblasts [104]. Increased gap junctional intercellular communication in primary human skin fibroblasts cells  were  observed  when  treated  with astaxanthin [99]. Astaxanthin inhibited cell death, cell proliferation and mammary tumors in chemically induced male/female rats and mice [105–109]. H. pluvialis extract inhibited the growth of human colon cancer cells by arresting cell cycle progression and promoting apoptosis reported by Palozza et al. [104]. Nitroastaxanthin and 15-nitroastaxanthin are the products of astaxanthin with peroxynitrite, 15-nitroastaxanthin anticancer properties were evaluated in a mouse model. Epstein-Barr virus and carcinogenesis in mouse skin papillomas were significantly inhibited by astaxanthin treatment [110].

     

    • Immuno-Modulation

     

    Immune system cells are very sensitive to free radical damage. The cell membrane contains poly unsaturated fatty acids (PUFA). Antioxidants in particular astaxanthin offer protection against free radical damage to preserve immune-system defenses. There are reports on astaxanthin and its effect on immunity in animals under laboratory conditions however clinical research is lacking in humans. Astaxanthin showed higher immuno-modulating effects in mouse model when compared  to  β-carotene [111]. Enhanced antibody production and decreased humoral immune response in older animals after dietary supplementation of astaxanthin was reported [111,112]. Astaxanthin produced immunoglobulins in human cells in a laboratory study [113]. Eight week-supplementation of astaxanthin in humans [72] resulted in increased blood levels of astaxanthin and improved activity of natural killer cells which targeted and destroyed cells infected with viruses. In this study, T and B cells were increased, DNA damage was low, and C-reactive protein (CRP) was significantly lower in the astaxanthin supplemented group [67,102,114]. Recent reports on astaxanthin biological activities are presented in Table 2.

     

    Table 2. Astaxanthin biological activities in in vitro and in vivo models.

     

             Biological Activities                    

    References           

    Antioxidant activity

    [14,15,17,115–120]

    Protection from UV rays

    [14]

    Anti-skin cancer

    [14,110,121]

    Anti-inflammatory

    [84,122–125]

    Anti-gastric activity

    [68,71]

    Anti-hepatoprotective

    [126]

    Anti-diabetes

    [90,127,128]

    Cardiovascular prevention

    [94,122,129,130]

    Immune response

    [72,114]

    Neuroprotection

    [131,132]

     

     

    9.  Safety and Dose of Astaxanthin

     

    Astaxanthin is safe, with no side effects when it is consumed with food. It is lipid soluble, accumulates in animal tissues  after  feeding  of  astaxanthin  to  rats  and  no  toxic  effects  were  found [15,17,133]. Excessive astaxanthin consumption leads to yellow to reddish pigmentation of the skin in animals. Astaxanthin is incorporated into fish feed, resulting in the fish skin becoming reddish in color. Antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase levels significantly increased in rats after oral dosage of astaxanthin [14,15]. A study reported that blood pressure (bp) was reduced in stroke prone rats and in hypertensive rats by feeding 50 mg/kg astaxanthin for five weeks and 14 days, respectively [134]. Astaxanthin was also shown significant protection against naproxen induced gastric, antral ulcer and inhibited lipid peroxidation levels in gastric mucosa [67,135]. Astaxanthin accumulation in eyes was observed when astaxanthin was fed to rats [136]. Astaxanthin extracted from Paracoccus carotinifaciens showed potential antioxidant and also anti-ulcer properties in murine models as reported by Murata et al. [137]. Astaxanthin bioavailability was increased with supplement of lipid based formulations [14,15,17,138]. Supratherapeutic concentrations of astaxanthin had no adverse effects on platelet, coagulation and fibrinolytic function [139]. Research has so far reported no significant side effects of astaxanthin consumption in animals and humans. These results support the safety of astaxanthin  for  future  clinical studies.

    It is recommended to administer astaxanthin with omega-3 rich seed oils such as chia, flaxseed,  fish, nutella, walnuts and almonds. The combination of astaxanthin (4–8 mg) with foods, soft gels and capsules and cream is available in the market. Recommended dose of astaxanthin is 2–4 mg/day. A study reported that no adverse effects were found with the administration of astaxanthin (6 mg/day) in adult human subjects [140]. Astaxanthin effects on human blood rheology were investigated in adult men  subjects  with  a  single-blind  method  after  administration  of  astaxanthin  at  6  mg/day  for   10 days [141]. Recent studies on astaxanthin dosage effects on human health benefits were presented  in Table 3.

     

    Table 3. Health benefits of astaxanthin in human subjects.

     

    Duration of Experiment

    Subjects in Humans

    Dosage (mg/day)

    Benefits of Astaxanthin

    References

    2 weeks

    Volunteers

    1.8, 3.6, 14.4 and 21.6

    Reduction of LDL oxidation

    [21]

    Single dose

    Middle aged male volunteers

    100

    Astaxanthin take up by VLDL chylomicrons

    [60]

    8 weeks

    Healthy females

    0.2 and 8

    Decreased plasma

    8-hydoxy-2′-deoxyguanosine and lowered in CRP levels

    [72]

    8 weeks

    Healthy adults

    6

    Assessed by blood pressure

    [140]

    10 days

    Healthy males

    6

    Improved blood rheology

    [141]

    12 weeks

    Healthy non-smoking finnish males

    8

    Decreased oxidation of fatty acids

    [142]

    12 months

    Age related macular degeneration

    4

    Improved central retinal dysfunction in age related macular degeneration

    [143]

     

     

    Table 3. Cont.

     

    12 weeks

    Middle aged/elderly

    12

    Improved Cog health battery scores

    [144]

    12 weeks

    Middle aged/elderly

    6

    Improved groton maze learning test scores

    [144]

    8 or 6 weeks

    Healthy female or male

    6

    Improved skin winkle, corneocyte layer, epidermis and dermis

    [145]

    2 weeks

    Disease (bilateral cataract)

    6

    Improved superoxide scavenging activity and lowered hydroperoxides in the human aqueous humor

    [146]

    LDL, Low-density lipoproteins, VLDL, Very low-density lipoprotein, CRP, C-reactive protein.

     

    10.  Commercial Applications of Astaxanthin

     

    In the present scenario, production of astaxanthin from natural sources has become one of the most successful activities in biotechnology. Astaxanthin has great demand in food, feed, nutraceutical and pharmaceutical applications. This has promoted major efforts to improve astaxanthin production from biological sources instead of synthetic ones. According to the current literature, astaxanthin is used in various commercial applications in the market. Astaxanthin products are available in the form of capsule, soft gel, tablet, powder, biomass, cream, energy drink, oil and extract in the market (Table 4). Some of the astaxanthin products were made with combination of other carotenoids, multivitamins, herbal extracts and omega-3, 6 fatty acids. Patent applications are available on astaxanthin for preventing bacterial infection, inflammation, vascular failure, cancer, cardiovascular diseases, inhibiting lipid peroxidation, reducing cell damage and body fat, and improving brain function and  skin thickness (Table 5). Astaxanthin containing microorganisms or animals find many applications in a wide range of commercial activities, the reason for which astaxanthin enriched microalgae  production can provide more attractive benefits.

     

    Table 4. Astaxanthin products from various companies and its use for various purposes.

     

    Brand Name

    Dosage form

    Ingredients

    Company Name

    Purpose

    Physician Formulas

    Soft gel/Tablets

    2 mg/4 mg-AX

    Physician formulas vitamin company

    Antioxidant

    Eyesight Rx

    Tablet

    AX, vitamin-C, plant extracts

    Physician formulas Vitamin company

    Vision function

    KriaXanthin

    Soft gel

    1.5 mg-AX, EPA, DHA

    Physician formulas vitamin company

    Antioxidant

    Astaxanthin Ultra

    Soft gel

    4 mg-AX

    AOR

    Cardiovascular health/gastrointestinal

    Astaxanthin Gold™

    Soft gel

    4 mg-AX

    Nutrigold

    Eye/joint/skin/immune health

    Best Astaxanthin

    Soft gel

    6 mg-AX, CX

    Bioastin

    Cell membrane/blood flow

    Dr.Mercola

    Capsules

    4 mg AX, 325 mg Omega-3 ALA

    Dr. Mercola premium supplements

    Aging/muscle

    Solgar

    Soft gel

    5 mg-AX

    Solgar global manufacture

    Healthy skin

    Astaxanthin

    Cream

    AX, herbal extracts

    True botanica

    Face moisturizing

     

     

    Table 4. Cont.

     

    astavita ex

    Capsules

    8 mg AX, T3

    Fuji Chemical Industry

    Agingcare

    astavita SPORT

    Capsules

    9 mg AX, T3 and zinc

    Fuji Chemical Industry

    Sports nutrition

    AstaREAL

    Oil, powder, water soluble, biomass

    AX, AX-esters

    Fuji Chemical Industry

    Soft gel, tablet, beverages, animal feed, capsules

    AstaTROL

    Oil

    AX

    Fuji Chemical Industry

    Cosmetics

    AstaFX

    Capsules

    AX

    Purity and products evidence based nutritional supplements

    Skin/cardiovascular function

    Pure Encapsulations

    Capsules

    AX

    Synergistic nutrition

    Antioxidant

    Zanthin Xp-3

    Soft gel capsules

    2 mg, 4 mg-AX

    Valensa

    Human body

    Micro Algae Super Food

    Soft gel

    4 mg AX

    Anumed intel biomed company

    heart/eye/joint

    (Information obtained from the respective company websites); AX, astaxanthin, AXE, astaxanthin esters, CX, canthaxanthin, DHA, docosahexaenoic acid, EPA, eicosapentaenoic acid, ALA, alpha linolenic acid, T3, tocotrienol.

     

    Table 5. Recent patent applications for astaxanthin.

     

    Patent No.

    Title

    Purpose

    References

    US20060217445

    Natural astaxanthin extract reduces DNA oxidation

    Reduce endogenous oxidative damage

    [147]

    US20070293568

    Neurocyte protective agent

    Neuroprotection

    [148]

    US20080234521

    Crystal forms of astaxanthin

    Nutritional dosage

    [149]

    US20080293679

    Use of carotenoids and carotenoid derivatives analogs for reduction/ inhibition of certain negative effects of COX inhibitors

    Inhibit of lipid peroxidation

    [150]

    US20090047304

    Composition for body fat reduction

    Inhibits body fat

    [151]

    US20090069417

    Carotenoid oxidation products as chemopreventive and chemotherapeutic agents

    Cancer prevention

    [152]

    US20090136469

    Formulation for oral administration with beneficial effects on the cardiovascular system

    Cardiovascular protection

    [153]

    US20090142431

    Algal and algal extract dietary supplement composition

    Dietary supplement

    [154]

    US20090297492

    Method for improving cognitive performance

    Improving brain function

    [155]

    US20100158984

    Encapsulates

    Capsules

    [156]

    US20100204523

    Method of preventing discoloration of carotenoid pigment and container used therefor

    Prevention of discoloration

    [157]

    US20100267838

    Pulverulent carotenoid preparation for colouring drinks

    Drinks

    [158]

    US20100291053

    Inflammatory disease treatment

    Preventing inflammatory disease

    [159]

    US20120004297

    Agent for alleviating vascular failure

    Preventing vascular failure

    [160]

    US20120114823

    Feed additive for improved pigment retention

    Fish feed

    [161]

    US20120238522

    Carotenoid containing compositions and methods

    Preventing bacterial infections

    [162]

    US20120253078

    Agent for improving carcass performance in finishing hogs

    Food supplements

    [163]

    US20130004582

    Composition and method to alleviate joint pain

    Reduced joint pain and symptoms of osteoarthritis

    [164]

    US20130108764

    Baked food produced from astaxanthin containing dough

    Astaxanthin used in baked food

    [165]

     

     

    11.  Conclusion

     

    The current research data on astaxanthin is encouraging and have resulted from well controlled  trials in in vitro and in vivo models. Astaxanthin showed potential effects on various diseases including cancers, hypertension, diabetes, cardiovascular, gastrointestinal, liver, neurodegenerative, and skin diseases. Its antioxidant properties are used against oxidative damage in diseased cells. Recently, our laboratory isolated and characterized astaxanthin and its esters from Haematococcus and checked their biological activities in in vitro and in vivo models, confirming that astaxanthin and its esters show potential biological activities in animal models. However, there is a lack of research on astaxanthin esters (mono-di) and their metabolic pathways in biological systems. Future research should focus on effects of astaxanthin esters on various biological activities and their uses in nutraceutical and pharmaceutical applications. Astaxanthin mono-diesters may increase biological activities better than the free form which can be easily absorbed into the metabolism. Further research requires to be investigated on their metabolic pathways and also molecular studies in in vitro and in vivo models for their use in commercial purposes.

     

    Acknowledgments

     

    The first author thanks the University of Malaya Research Grant (UMRG RP001i-13SUS), University of Malaya, Kuala Lumpur, Malaysia for providing financial support for this project.

     

    Conflicts of Interest

     

    The authors declare no conflict of interest.

     

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  • Natural Standard

    By Natural Medicine Journal

    Related Terms

     

    3,3'-Dihydroxy-4,4'-diketo-beta-carotene, 3,3'-dihydroxy- beta,beta'-caroten-4,4'-dione, 3R,3'R-astaxanthin, 3R,3'S- astaxanthin, 3S,3'S-astaxanthin, Agrobacterium

    aurantiacum, alpha-carotene, Antarctic krill, AST, AstaCarox®, AstaFactor® Rejuvenating Formula, AstaFactor® Sports Formula, Astavita AstaREAL®, astaxanthin diester, astaxanthin dilysinate tetrahydrochloride, astaxanthin-amino acid conjugate, astaxanthine, Astaxin®, ASX, Atlantic salmon, basidiomycete yeast, beta-carotene, Botryococcus braunii, canthaxanthin, canthoxanthin, Cardax®, carotenoid, CDX-085, Chlorella zofingiensis, Chlorococcum spp., crayfish, crustaceans, DDA, disodium disuccinate astaxanthin, E161j, Euphausia superba, fatty acids, flamingo, gamma-tocopherol, green microalgae, Haematococcus algae extract, Haematococcus pluvialis, homochiral (3S,3'S)-astaxanthin, krill, lutein, lycopene, meso-3R,3'S isomer, meso-astaxanthin, microalgae, nonesterified astaxanthin, non-provitamin A carotenoid, omega-3 fatty acids, ovoester, Phaffia rhodozyma, propolis, quail, red carotenoid, retinoid, salmon, shrimp, sockeye salmon, storks, terpenoids, tetrahydrochloride dilysine astaxanthin salt, tomato, trout, wild salmon, Xanthophyllomyces dendrorhous, xanthophylls.

     

    Background

     

    Astaxanthin is a naturally occurring carotenoid found in nature primarily in marine organisms such as microalgae, salmon, trout, krill, shrimp, crayfish, and crustaceans. The green microalga Haematococcus pluvialis is considered the richest source of astaxanthin. Other microalgae, such as Chlorella zofingiensis, Chlorococcum spp., and Botryococcus braunii, also contain astaxanthin. It may also be found in the feathers of birds, such as quail, flamingo, and storks, as well as in propolis, the resinous substance collected by bees.

    Carotenoids are well known for their therapeutic benefits in the aging process and various diseases, because of their antioxidant properties. Astaxanthin is a xanthophyll carotenoid like lutein, zeaxanthin, and cryptoxanthin, which do not convert to vitamin A.

    According to a review, carotenoids are of interest based on their beneficial mechanisms of action for cancers, cardiovascular disease, age-related macular degeneration, and cataract formation. Numerous studies support the use of astaxanthin as a potent antioxidant that may be beneficial in decreasing the risks of certain chronic diseases. It may also reduce oxidative stress in the nervous system, reducing the risk of neurodegenerative diseases. Additionally, astaxanthin has well-documented anti-inflammatory and immune-stimulating effects.

    Human trials have been conducted in disorders such as carpal tunnel syndrome, rheumatoid arthritis, dyspepsia (with or withoutHelicobacter pylori infection), hyperlipidemia, male infertility, and skin conditions, and regarding exercise capacity, muscle soreness, and transplants.

    However, results have been mixed, and more research is needed in these areas before any firm conclusions can be drawn.

     

    Scientific Evidence

     

    Uses

    These uses have been tested in humans or animals. Safety and effectiveness have not always been proven. Some of these conditions are potentially serious, and should be evaluated by a qualified healthcare provider.

     

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    Antioxidant

     

     

     

     

     

     

     

     

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    Evidence suggests that astaxanthin may have antioxidant activity. Additional research is needed to confirm these results.

    B

    Carpal tunnel syndrome

    Preliminary research suggests that astaxanthin, as part of a multi-ingredient antioxidant supplement, may reduce pain and duration associated with carpal tunnel syndrome.

    However, larger studies are warranted before a conclusion can be drawn.

     

    C

    Dyspepsia

    Limited evidence suggests that astaxanthin may be beneficial in dyspepsia. Additional evidence is warranted before a conclusion can be drawn.

     

    C

    Exercise capacity

    High-quality evidence supporting the use of astaxanthin to improve exercise capacity is lacking. More studies are warranted before a conclusion can be drawn.

     

    C

    High cholesterol

    High-quality evidence supporting the use of astaxanthin for high cholesterol is lacking. More studies are warranted before a conclusion can be drawn.

     

    C

    Macular degeneration

    Preliminary research suggests that astaxanthin, as part of a multi-ingredient supplement, may benefit patients with macular degeneration. However, studies evaluating astaxanthin alone are warranted before a conclusion can be drawn.

     

    C

    Male infertility

    Limited evidence suggests that astaxanthin may be beneficial in male infertility. Additional evidence is warranted before a conclusion can be drawn.

     

    C

    Menopausal symptoms

    According to preliminary research, a combination product containing astaxanthin was found to reduce climacteric symptoms in women with menopause. Studies evaluating astaxanthin alone are needed before a conclusion can be drawn.

     

    C

    Muscle soreness

    Evidence supporting the use of astaxanthin for muscle soreness is lacking. More studies are warranted before a conclusion can be drawn.

     

    C

    Rheumatoid arthritis

    According to preliminary research, astaxanthin may be beneficial in alleviating pain and improving the ability to perform daily activity in patients with rheumatoid arthritis. However, larger studies are warranted before a conclusion can be drawn.

     

    C

    Skin conditions

    According to preliminary research, astaxanthin may help reduce fine lines and wrinkles and improve skin elasticity and moisture content. More studies are warranted before a conclusion can be drawn.

     

    C

    Transplants

    There is an ongoing study being conducted assessing the effects of astaxanthin on vascular structure, oxidative stress, and inflammation in renal transplant patients. Results of this trial are pending.

     

    C

     

    Tradition/Theory

     

    The below uses are based on tradition, scientific theories, or limited research. They often have not been thoroughly tested in humans, and safety and effectiveness have not always been proven. Some of these conditions are potentially serious, and should be evaluated by a qualified healthcare provider. There may be other proposed uses that are not listed below.

    Alzheimer's disease, anti-inflammatory, antimicrobial, antiviral, anxiety, arthritis, asthma, atherosclerosis (prevention), autoimmune diseases, back pain (chronic), benign prostate hyperplasia, cancer, canker sores, cardiovascular disease, cataracts, chronic illness, dementia, depression, diabetes, diabetic neuropathy, eye problems, gastrointestinal disorders, hepatitis, hormonal effects, hypertension, immune stimulant, ischemic injury, leukemia, liver disorders, mitochondrial diseases, neuroprotection, obesity, panic disorder, Parkinson's disease, photoprotection, renal failure, stroke, thrombosis, toxicity (iron-chelate; drug-induced cardiotoxicity), vascular disorders.

     

    Dosing

     

    The below doses are based on scientific research, publications, traditional use, or expert opinion. Many herbs and supplements have not been thoroughly tested, and safety and effectiveness may not be proven. Brands may be made differently, with variable ingredients, even within the same brand. The below doses may not apply to all products. You should read product labels, and discuss doses with a qualified healthcare provider before starting therapy.

     

    Adults (18 years and older)

     

    As an antioxidant, in general, manufacturers recommend taking 4-8 milligrams of astaxanthin by mouth 2-3 times daily with meals. Clinical evidence is lacking.

    For dyspepsia, 40 milligrams of astaxanthin (AstaCarox®) has been taken by mouth daily in divided doses for four weeks.

    For exercise capacity, the manufacturers of Xanthin® recommend taking one capsule by mouth (each containing eight milligrams of astaxanthin) before and after physical activity. Four milligrams of astaxanthin has also been taken by mouth in the morning with food.

    For high cholesterol, 6, 12, and 18 milligrams of astaxanthin (AstaREAL® Astaxanthin) has been taken by mouth daily for 12 weeks. In healthy human subjects, 3.6, 7.2, and 14.4 milligrams has been administered in a beverage daily for two weeks.

    For male infertility, 16 milligrams of astaxanthin (AstaCarox®) has been taken by mouth daily for three months.

    For skin conditions, two milligrams of astaxanthin (Astavita AstaREAL® Astaxanthin; each capsule containing two milligrams of astaxanthin derived from Haematococcus plubialis microalgae) has been taken by mouth twice daily with breakfast and dinner for six weeks. According to secondary sources, four milligrams of astaxanthin (BioAstin®) daily for two weeks may prevent sunburn.

    For transplant (renal), 12 milligrams of astaxanthin (BioAstin®; four milligram tablets taken by mouth three times daily) has been used for one year.

    Note: Various seafoods contain the astaxanthin pigment. A standard serving portion of four ounces of Atlantic salmon contains from 0.5 to 1.1 milligrams of astaxanthin, whereas the same amount of sockeye salmon may contain 4.5 milligrams of astaxanthin.

    Children (under 18 years old)

     

    There is no proven safe or effective dose for astaxanthin in children.

     

    Safety

     

    The U.S. Food and Drug Administration does not strictly regulate herbs and supplements. There is no guarantee of strength, purity or safety of products, and effects may vary. You should always read product labels. If you have a medical condition, or are taking other drugs, herbs, or supplements, you should speak with a qualified healthcare provider before starting a newtherapy. Consult a healthcare provider immediately if you experience side effects.

     

    Allergies

     

    Avoid with known allergy or hypersensitivity to astaxanthin or related carotenoids, including canthaxanthin, or hypersensitivity to an astaxanthin source, such as Haematococcus pluvialis.

    Side Effects and Warnings

     

    Astaxanthin may affect bleeding. Caution is advised in patients with bleeding disorders or those taking drugs that may affect bleeding. Dosing adjustments may be necessary.

    Astaxanthin may lower blood sugar levels. Caution is advised in patients with diabetes or hypoglycemia, and in those taking drugs, herbs, or supplements that affect blood sugar. Blood glucose levels may need to be monitored by a qualified healthcare professional, including a pharmacist. Medication adjustments may be necessary.

    Astaxanthin may cause low blood pressure. Caution is advised in patients taking drugs, herbs, or supplements that lower blood pressure.

    Use cautiously in patients with taking certain drugs, herbs and supplements metabolized by the liver's cytochrome P450 enzyme system. Taking astaxanthin with these drugs may cause the levels of these drugs to be decreased in the blood and may reduce the intended effects. Patients taking any medications should check the package insert and speak with a qualified healthcare professional, including a pharmacist, about possible interactions.

    Use cautiously in patients with hormone disorders or those using agents that affect hormones, particularly 5-alpha-reductase inhibitors, as astaxanthin may inhibit 5-alpha-reductase, thereby

     

    inhibiting the conversion of testosterone to dihydrotestosterone (DHT). Theoretically, adverse effects related to 5-alpha-reductase inhibitors, such as decreased libido, gynecomastia, decreased semen quantity during ejaculation, impotence, increased skin pigmentation, hair growth, weight gain, and depressed mood, may occur.

    Use cautiously in patients with autoimmune disorders or those using immunosuppressants, as astaxanthin has been shown to enhance immune function and theoretically may interfere with immunosuppressive therapy. Although astaxanthin has been found to stimulate the immune system, in clinical research, astaxanthin was found to lower eosinophil levels.

    Use cautiously in patients with hypocalcemia, osteoporosis, or parathyroid disorders, as astaxanthin may lower serum calcium levels.

    Use cautiously in patients using beta-carotene, as astaxanthin may alter beta-carotene conversion.

    Use cautiously in women who are pregnant or might become pregnant, as astaxanthin may inhibit 5-alpha reductase.

    Astaxanthin may also cause severe abdominal pain and aplastic anemia.

    Avoid in patients with known allergy or hypersensitivity to astaxanthin or related carotenoids, including canthaxanthin, or in those with hypersensitivity to an astaxanthin source, such as Haematococcus pluvialis.

    Avoid in patients with known hypersensitivity to 5-alpha-reductase inhibitors.

     

    Pregnancy and Breastfeeding

     

    Use cautiously in women who are pregnant or might become pregnant, as astaxanthin may inhibit 5-alpha reductase. Astaxanthin is not suggested in breastfeeding women, due to a lack of safety data.

     

     

     

    Interactions

     

    Most herbs and supplements have not been thoroughly tested for interactions with other herbs, supplements, drugs, or foods. The interactions listed beloware based on reports in scientific publications, laboratory experiments, or traditional use. You should always read product labels. If you have a medical condition, or are taking other drugs, herbs, or supplements, you should speak with a qualified healthcare provider before starting a newtherapy.

     

    Interactions with Drugs

     

    Astaxanthin may increase the risk of bleeding or blood clotting when taken with drugs that increase such risks. Some examples include aspirin, anticoagulants (blood thinners) such as warfarin (Coumadin®) or heparin, antiplatelet drugs such as clopidogrel (Plavix®), and nonsteroidal anti-inflammatory drugs such as ibuprofen (Motrin®, Advil®) or naproxen (Naprosyn®, Aleve®).

    Astaxanthin may lower blood sugar levels. Caution is advised when using medications that may also lower blood sugar. Patients taking insulin or drugs for diabetes by mouth should be monitored closely by a qualified healthcare professional, including a pharmacist. Medication adjustments may be necessary.

    Astaxanthin may cause low blood pressure. Caution is advised in patients taking drugs, herbs, or supplements that lower blood pressure.

    Astaxanthin may interfere with the way the body processes certain drugs using the liver's cytochrome P450 enzyme system. As a result, the levels of these drugs may be decreased in the blood and may reduce the intended effects. Patients taking any medications should check the package insert and speak with a qualified healthcare professional, including a pharmacist, about possible interactions.

    Astaxanthin may interact with 5-alpha-reductase inhibitors, calcium salts, diabetes drugs, drugs that affect bleeding, heart medications, hormonal agents, immunosuppressants, and rofecoxib.

    Interactions with Herbs and Dietary Supplements

     

    Astaxanthin may increase the risk of bleeding or blood clotting when taken with herbs or supplements that are believed to increase such risks. Multiple cases of bleeding have been reported with the use of Ginkgo biloba, and fewer cases with garlic and saw palmetto. Numerous other agents may theoretically increase the risk of bleeding or blood clotting, although this has not been proven in most cases.

    Astaxanthin may lower blood sugar levels. Caution is advised when using herbs or supplements

     

    that may also lower blood sugar. Blood glucose levels may require monitoring, and doses may need adjustment.

    Astaxanthin may cause low blood pressure. Caution is advised in patients taking drugs, herbs, or supplements that lower blood pressure.

    Astaxanthin may interfere with the way the body processes certain herbs or supplements using the liver's cytochrome P450 enzyme system. As a result, the levels of other herbs or supplements may become too low in the blood. It may also alter the effects that other herbs or supplements potentially may have on the cytochrome P450 system.

    Astaxanthin may interact with calcium (and foods containing calcium), carotenoids (and foods containing carotenoids), herbs and supplements for the heart, hormonal herbs and supplements, immunosuppressants, and saw palmetto.

     

    Author Information

     

    This information is based on a systematic review of scientific literature edited and peer-reviewed by contributors to the Natural Standard Research Collaboration (www.naturalstandard.com).

     

    References

     

    • Andersen, P., Holck, S., Kupcinskas, L., Kiudelis, G., Jonaitis, L., Janciauskas, D., Permin, H., and Wadstrom, T. Gastric inflammatory markers and interleukins in patients with functional dyspepsia treated with astaxanthin. FEMSImmunol.Med Microbiol. 2007;50(2):244- 248.
    • Belcaro, , Cesarone, M. R., Cornelli, U., and Dugall, M. MF Afragil(R) in the treatment of  34 menopause symptoms: a pilot   study.

    Panminerva Med. 2010;52(2 Suppl 1):49-54.

    • Comhaire, H., El Garem, Y., Mahmoud, A., Eertmans, F., and Schoonjans, F. Combined conventional/antioxidant "Astaxanthin" treatment for male infertility: a double blind, randomized trial. Asian J Androl. 2005;7(3):257-262.
    • Fassett, G. and Coombes, J. S. Astaxanthin: a potential therapeutic agent in cardiovascular disease. Mar.Drugs. 2011;9(3):447-465. 5 Horie, S., Okuda, C., Yamashita, T., Watanabe, K., Kuramochi, K., Hosokawa, M., Takeuchi, T., Kakuda, M., Miyashita, K., Sugawara, F., Yoshida, H., and Mizushina, Y. Purified canola lutein selectively inhibits specific isoforms of mammalian DNA polymerases and reduces inflammatory response. Lipids. 2010;45(8):713-721.
    • Kupcinskas, , Lafolie, P., Lignell, A., Kiudelis, G., Jonaitis, L., Adamonis, K., Andersen, L. P., and Wadstrom, T. Efficacy of the natural antioxidant astaxanthin in the treatment of functional dyspepsia in patients with or without Helicobacter pylori infection: A prospective, randomized, double blind, and placebo-controlled study. Phytomedicine. 2008;15(6-7):391-399.
    • Lignell, Å. Medicament for improvement of duration of muscle function or treatment of muscle disorders or 1999;Patent Cooperation Treaty application #9911251
    • Liu, X. and Osawa, Astaxanthin protects neuronal cells against oxidative damage and is a potent candidate for brain food. Forum Nutr. 2009;61:129-135.
    • Nakagawa, , Kiko, T., Miyazawa, T., Carpentero, Burdeos G., Kimura, F., Satoh, A., and Miyazawa, T. Antioxidant effect of astaxanthin on phospholipid peroxidation in human erythrocytes. Br J Nutr. 2011;105(11):1563-1571.
    • Parisi, , Tedeschi, M., Gallinaro, G., Varano, M., Saviano, S., and Piermarocchi, S. Carotenoids and antioxidants in age-related maculopathy italian study: multifocal electroretinogrammodifications after 1 year. Ophthalmology. 2008;115(2):324-333.
    • Pashkow, J., Watumull, D. G., and Campbell, C. L. Astaxanthin: a novel potential treatment for oxidative stress and inflammation in cardiovascular disease. Am J Cardiol. 5-22-2008;101(10A):58D-68D.
    • Serebruany, , Malinin, A., Goodin, T., and Pashkow, F. The in vitro effects of Xancor, a synthetic astaxanthine derivative, on hemostatic biomarkers in aspirin-naive and aspirin-treated subjects with multiple risk factors for vascular disease. Am J Ther.

    2010;17(2):125-132.

    • Yamashita The effect of a dietary supplement containing astaxanthin on skin condition. Carotenoid Sci. 2006;10:91-95.
    • Yoshida, , Yanai, H., Ito, K., Tomono, Y., Koikeda, T., Tsukahara, H., and Tada, N. Administration of natural astaxanthin increases serumHDL-cholesterol and adiponectin in subjects with mild hyperlipidemia. Atherosclerosis. 2010;209(2):520-523.
    • Yuan, P., Peng, J., Yin, K., and Wang, J. H. Potential health-promoting effects of astaxanthin: a high-value carotenoid mostly from microalgae. Mol Nutr Food Res. 2011;55(1):150-165.
     
       

     

     

     
       

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

    Related Terms

     

    3,3'-Dihydroxy-4,4'-diketo-beta-carotene, 3,3'-dihydroxy- beta,beta'-caroten-4,4'-dione, 3R,3'R-astaxanthin, 3R,3'S- astaxanthin, 3S,3'S-astaxanthin, Agrobacterium

    aurantiacum, alpha-carotene, Antarctic krill, AST, AstaCarox®, AstaFactor® Rejuvenating Formula, AstaFactor® Sports Formula, Astavita AstaREAL®, astaxanthin diester, astaxanthin dilysinate tetrahydrochloride, astaxanthin-amino acid conjugate, astaxanthine, Astaxin®, ASX, Atlantic salmon, basidiomycete yeast, beta-carotene, Botryococcus braunii, canthaxanthin, canthoxanthin, Cardax®, carotenoid, CDX-085, Chlorella zofingiensis, Chlorococcum spp., crayfish, crustaceans, DDA, disodium disuccinate astaxanthin, E161j, Euphausia superba, fatty acids, flamingo, gamma-tocopherol, green microalgae, Haematococcus algae extract, Haematococcus pluvialis, homochiral (3S,3'S)-astaxanthin, krill, lutein, lycopene, meso-3R,3'S isomer, meso-astaxanthin, microalgae, nonesterified astaxanthin, non-provitamin A carotenoid, omega-3 fatty acids, ovoester, Phaffia rhodozyma, propolis, quail, red carotenoid, retinoid, salmon, shrimp, sockeye salmon, storks, terpenoids, tetrahydrochloride dilysine astaxanthin salt, tomato, trout, wild salmon, Xanthophyllomyces dendrorhous, xanthophylls.

     

    Background

     

    Astaxanthin is a naturally occurring carotenoid found in nature primarily in marine organisms such as microalgae, salmon, trout, krill, shrimp, crayfish, and crustaceans. The green microalga Haematococcus pluvialis is considered the richest source of astaxanthin. Other microalgae, such as Chlorella zofingiensis, Chlorococcum spp., and Botryococcus braunii, also contain astaxanthin. It may also be found in the feathers of birds, such as quail, flamingo, and storks, as well as in propolis, the resinous substance collected by bees.

    Carotenoids are well known for their therapeutic benefits in the aging process and various diseases, because of their antioxidant properties. Astaxanthin is a xanthophyll carotenoid like lutein, zeaxanthin, and cryptoxanthin, which do not convert to vitamin A.

    According to a review, carotenoids are of interest based on their beneficial mechanisms of action for cancers, cardiovascular disease, age-related macular degeneration, and cataract formation. Numerous studies support the use of astaxanthin as a potent antioxidant that may be beneficial in decreasing the risks of certain chronic diseases. It may also reduce oxidative stress in the nervous system, reducing the risk of neurodegenerative diseases. Additionally, astaxanthin has well-documented anti-inflammatory and immune-stimulating effects.

    Human trials have been conducted in disorders such as carpal tunnel syndrome, rheumatoid arthritis, dyspepsia (with or withoutHelicobacter pylori infection), hyperlipidemia, male infertility, and skin conditions, and regarding exercise capacity, muscle soreness, and transplants.

    However, results have been mixed, and more research is needed in these areas before any firm conclusions can be drawn.

     

    Scientific Evidence

     

    Uses

    These uses have been tested in humans or animals. Safety and effectiveness have not always been proven. Some of these conditions are potentially serious, and should be evaluated by a qualified healthcare provider.

     

    Grade*

    Antioxidant

     

     

     

     

     

     

     

     

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    converted by W eb 2PDFConvert.com

     

    Evidence suggests that astaxanthin may have antioxidant activity. Additional research is needed to confirm these results.

    B

    Carpal tunnel syndrome

    Preliminary research suggests that astaxanthin, as part of a multi-ingredient antioxidant supplement, may reduce pain and duration associated with carpal tunnel syndrome.

    However, larger studies are warranted before a conclusion can be drawn.

     

    C

    Dyspepsia

    Limited evidence suggests that astaxanthin may be beneficial in dyspepsia. Additional evidence is warranted before a conclusion can be drawn.

     

    C

    Exercise capacity

    High-quality evidence supporting the use of astaxanthin to improve exercise capacity is lacking. More studies are warranted before a conclusion can be drawn.

     

    C

    High cholesterol

    High-quality evidence supporting the use of astaxanthin for high cholesterol is lacking. More studies are warranted before a conclusion can be drawn.

     

    C

    Macular degeneration

    Preliminary research suggests that astaxanthin, as part of a multi-ingredient supplement, may benefit patients with macular degeneration. However, studies evaluating astaxanthin alone are warranted before a conclusion can be drawn.

     

    C

    Male infertility

    Limited evidence suggests that astaxanthin may be beneficial in male infertility. Additional evidence is warranted before a conclusion can be drawn.

     

    C

    Menopausal symptoms

    According to preliminary research, a combination product containing astaxanthin was found to reduce climacteric symptoms in women with menopause. Studies evaluating astaxanthin alone are needed before a conclusion can be drawn.

     

    C

    Muscle soreness

    Evidence supporting the use of astaxanthin for muscle soreness is lacking. More studies are warranted before a conclusion can be drawn.

     

    C

    Rheumatoid arthritis

    According to preliminary research, astaxanthin may be beneficial in alleviating pain and improving the ability to perform daily activity in patients with rheumatoid arthritis. However, larger studies are warranted before a conclusion can be drawn.

     

    C

    Skin conditions

    According to preliminary research, astaxanthin may help reduce fine lines and wrinkles and improve skin elasticity and moisture content. More studies are warranted before a conclusion can be drawn.

     

    C

    Transplants

    There is an ongoing study being conducted assessing the effects of astaxanthin on vascular structure, oxidative stress, and inflammation in renal transplant patients. Results of this trial are pending.

     

    C

     

    Tradition/Theory

     

    The below uses are based on tradition, scientific theories, or limited research. They often have not been thoroughly tested in humans, and safety and effectiveness have not always been proven. Some of these conditions are potentially serious, and should be evaluated by a qualified healthcare provider. There may be other proposed uses that are not listed below.

    Alzheimer's disease, anti-inflammatory, antimicrobial, antiviral, anxiety, arthritis, asthma, atherosclerosis (prevention), autoimmune diseases, back pain (chronic), benign prostate hyperplasia, cancer, canker sores, cardiovascular disease, cataracts, chronic illness, dementia, depression, diabetes, diabetic neuropathy, eye problems, gastrointestinal disorders, hepatitis, hormonal effects, hypertension, immune stimulant, ischemic injury, leukemia, liver disorders, mitochondrial diseases, neuroprotection, obesity, panic disorder, Parkinson's disease, photoprotection, renal failure, stroke, thrombosis, toxicity (iron-chelate; drug-induced cardiotoxicity), vascular disorders.

     

    Dosing

     

    The below doses are based on scientific research, publications, traditional use, or expert opinion. Many herbs and supplements have not been thoroughly tested, and safety and effectiveness may not be proven. Brands may be made differently, with variable ingredients, even within the same brand. The below doses may not apply to all products. You should read product labels, and discuss doses with a qualified healthcare provider before starting therapy.

     

    Adults (18 years and older)

     

    As an antioxidant, in general, manufacturers recommend taking 4-8 milligrams of astaxanthin by mouth 2-3 times daily with meals. Clinical evidence is lacking.

    For dyspepsia, 40 milligrams of astaxanthin (AstaCarox®) has been taken by mouth daily in divided doses for four weeks.

    For exercise capacity, the manufacturers of Xanthin® recommend taking one capsule by mouth (each containing eight milligrams of astaxanthin) before and after physical activity. Four milligrams of astaxanthin has also been taken by mouth in the morning with food.

    For high cholesterol, 6, 12, and 18 milligrams of astaxanthin (AstaREAL® Astaxanthin) has been taken by mouth daily for 12 weeks. In healthy human subjects, 3.6, 7.2, and 14.4 milligrams has been administered in a beverage daily for two weeks.

    For male infertility, 16 milligrams of astaxanthin (AstaCarox®) has been taken by mouth daily for three months.

    For skin conditions, two milligrams of astaxanthin (Astavita AstaREAL® Astaxanthin; each capsule containing two milligrams of astaxanthin derived from Haematococcus plubialis microalgae) has been taken by mouth twice daily with breakfast and dinner for six weeks. According to secondary sources, four milligrams of astaxanthin (BioAstin®) daily for two weeks may prevent sunburn.

    For transplant (renal), 12 milligrams of astaxanthin (BioAstin®; four milligram tablets taken by mouth three times daily) has been used for one year.

    Note: Various seafoods contain the astaxanthin pigment. A standard serving portion of four ounces of Atlantic salmon contains from 0.5 to 1.1 milligrams of astaxanthin, whereas the same amount of sockeye salmon may contain 4.5 milligrams of astaxanthin.

    Children (under 18 years old)

     

    There is no proven safe or effective dose for astaxanthin in children.

     

    Safety

     

    The U.S. Food and Drug Administration does not strictly regulate herbs and supplements. There is no guarantee of strength, purity or safety of products, and effects may vary. You should always read product labels. If you have a medical condition, or are taking other drugs, herbs, or supplements, you should speak with a qualified healthcare provider before starting a newtherapy. Consult a healthcare provider immediately if you experience side effects.

     

    Allergies

     

    Avoid with known allergy or hypersensitivity to astaxanthin or related carotenoids, including canthaxanthin, or hypersensitivity to an astaxanthin source, such as Haematococcus pluvialis.

    Side Effects and Warnings

     

    Astaxanthin may affect bleeding. Caution is advised in patients with bleeding disorders or those taking drugs that may affect bleeding. Dosing adjustments may be necessary.

    Astaxanthin may lower blood sugar levels. Caution is advised in patients with diabetes or hypoglycemia, and in those taking drugs, herbs, or supplements that affect blood sugar. Blood glucose levels may need to be monitored by a qualified healthcare professional, including a pharmacist. Medication adjustments may be necessary.

    Astaxanthin may cause low blood pressure. Caution is advised in patients taking drugs, herbs, or supplements that lower blood pressure.

    Use cautiously in patients with taking certain drugs, herbs and supplements metabolized by the liver's cytochrome P450 enzyme system. Taking astaxanthin with these drugs may cause the levels of these drugs to be decreased in the blood and may reduce the intended effects. Patients taking any medications should check the package insert and speak with a qualified healthcare professional, including a pharmacist, about possible interactions.

    Use cautiously in patients with hormone disorders or those using agents that affect hormones, particularly 5-alpha-reductase inhibitors, as astaxanthin may inhibit 5-alpha-reductase, thereby

     

    inhibiting the conversion of testosterone to dihydrotestosterone (DHT). Theoretically, adverse effects related to 5-alpha-reductase inhibitors, such as decreased libido, gynecomastia, decreased semen quantity during ejaculation, impotence, increased skin pigmentation, hair growth, weight gain, and depressed mood, may occur.

    Use cautiously in patients with autoimmune disorders or those using immunosuppressants, as astaxanthin has been shown to enhance immune function and theoretically may interfere with immunosuppressive therapy. Although astaxanthin has been found to stimulate the immune system, in clinical research, astaxanthin was found to lower eosinophil levels.

    Use cautiously in patients with hypocalcemia, osteoporosis, or parathyroid disorders, as astaxanthin may lower serum calcium levels.

    Use cautiously in patients using beta-carotene, as astaxanthin may alter beta-carotene conversion.

    Use cautiously in women who are pregnant or might become pregnant, as astaxanthin may inhibit 5-alpha reductase.

    Astaxanthin may also cause severe abdominal pain and aplastic anemia.

    Avoid in patients with known allergy or hypersensitivity to astaxanthin or related carotenoids, including canthaxanthin, or in those with hypersensitivity to an astaxanthin source, such as Haematococcus pluvialis.

    Avoid in patients with known hypersensitivity to 5-alpha-reductase inhibitors.

     

    Pregnancy and Breastfeeding

     

    Use cautiously in women who are pregnant or might become pregnant, as astaxanthin may inhibit 5-alpha reductase. Astaxanthin is not suggested in breastfeeding women, due to a lack of safety data.

     

     

     

    Interactions

     

    Most herbs and supplements have not been thoroughly tested for interactions with other herbs, supplements, drugs, or foods. The interactions listed beloware based on reports in scientific publications, laboratory experiments, or traditional use. You should always read product labels. If you have a medical condition, or are taking other drugs, herbs, or supplements, you should speak with a qualified healthcare provider before starting a newtherapy.

     

    Interactions with Drugs

     

    Astaxanthin may increase the risk of bleeding or blood clotting when taken with drugs that increase such risks. Some examples include aspirin, anticoagulants (blood thinners) such as warfarin (Coumadin®) or heparin, antiplatelet drugs such as clopidogrel (Plavix®), and nonsteroidal anti-inflammatory drugs such as ibuprofen (Motrin®, Advil®) or naproxen (Naprosyn®, Aleve®).

    Astaxanthin may lower blood sugar levels. Caution is advised when using medications that may also lower blood sugar. Patients taking insulin or drugs for diabetes by mouth should be monitored closely by a qualified healthcare professional, including a pharmacist. Medication adjustments may be necessary.

    Astaxanthin may cause low blood pressure. Caution is advised in patients taking drugs, herbs, or supplements that lower blood pressure.

    Astaxanthin may interfere with the way the body processes certain drugs using the liver's cytochrome P450 enzyme system. As a result, the levels of these drugs may be decreased in the blood and may reduce the intended effects. Patients taking any medications should check the package insert and speak with a qualified healthcare professional, including a pharmacist, about possible interactions.

    Astaxanthin may interact with 5-alpha-reductase inhibitors, calcium salts, diabetes drugs, drugs that affect bleeding, heart medications, hormonal agents, immunosuppressants, and rofecoxib.

    Interactions with Herbs and Dietary Supplements

     

    Astaxanthin may increase the risk of bleeding or blood clotting when taken with herbs or supplements that are believed to increase such risks. Multiple cases of bleeding have been reported with the use of Ginkgo biloba, and fewer cases with garlic and saw palmetto. Numerous other agents may theoretically increase the risk of bleeding or blood clotting, although this has not been proven in most cases.

    Astaxanthin may lower blood sugar levels. Caution is advised when using herbs or supplements

     

    that may also lower blood sugar. Blood glucose levels may require monitoring, and doses may need adjustment.

    Astaxanthin may cause low blood pressure. Caution is advised in patients taking drugs, herbs, or supplements that lower blood pressure.

    Astaxanthin may interfere with the way the body processes certain herbs or supplements using the liver's cytochrome P450 enzyme system. As a result, the levels of other herbs or supplements may become too low in the blood. It may also alter the effects that other herbs or supplements potentially may have on the cytochrome P450 system.

    Astaxanthin may interact with calcium (and foods containing calcium), carotenoids (and foods containing carotenoids), herbs and supplements for the heart, hormonal herbs and supplements, immunosuppressants, and saw palmetto.

     

    Author Information

     

    This information is based on a systematic review of scientific literature edited and peer-reviewed by contributors to the Natural Standard Research Collaboration (www.naturalstandard.com).

     

    References

     

    • Andersen, P., Holck, S., Kupcinskas, L., Kiudelis, G., Jonaitis, L., Janciauskas, D., Permin, H., and Wadstrom, T. Gastric inflammatory markers and interleukins in patients with functional dyspepsia treated with astaxanthin. FEMSImmunol.Med Microbiol. 2007;50(2):244- 248.
    • Belcaro, , Cesarone, M. R., Cornelli, U., and Dugall, M. MF Afragil(R) in the treatment of  34 menopause symptoms: a pilot   study.

    Panminerva Med. 2010;52(2 Suppl 1):49-54.

    • Comhaire, H., El Garem, Y., Mahmoud, A., Eertmans, F., and Schoonjans, F. Combined conventional/antioxidant "Astaxanthin" treatment for male infertility: a double blind, randomized trial. Asian J Androl. 2005;7(3):257-262.
    • Fassett, G. and Coombes, J. S. Astaxanthin: a potential therapeutic agent in cardiovascular disease. Mar.Drugs. 2011;9(3):447-465. 5 Horie, S., Okuda, C., Yamashita, T., Watanabe, K., Kuramochi, K., Hosokawa, M., Takeuchi, T., Kakuda, M., Miyashita, K., Sugawara, F., Yoshida, H., and Mizushina, Y. Purified canola lutein selectively inhibits specific isoforms of mammalian DNA polymerases and reduces inflammatory response. Lipids. 2010;45(8):713-721.
    • Kupcinskas, , Lafolie, P., Lignell, A., Kiudelis, G., Jonaitis, L., Adamonis, K., Andersen, L. P., and Wadstrom, T. Efficacy of the natural antioxidant astaxanthin in the treatment of functional dyspepsia in patients with or without Helicobacter pylori infection: A prospective, randomized, double blind, and placebo-controlled study. Phytomedicine. 2008;15(6-7):391-399.
    • Lignell, Å. Medicament for improvement of duration of muscle function or treatment of muscle disorders or 1999;Patent Cooperation Treaty application #9911251
    • Liu, X. and Osawa, Astaxanthin protects neuronal cells against oxidative damage and is a potent candidate for brain food. Forum Nutr. 2009;61:129-135.
    • Nakagawa, , Kiko, T., Miyazawa, T., Carpentero, Burdeos G., Kimura, F., Satoh, A., and Miyazawa, T. Antioxidant effect of astaxanthin on phospholipid peroxidation in human erythrocytes. Br J Nutr. 2011;105(11):1563-1571.
    • Parisi, , Tedeschi, M., Gallinaro, G., Varano, M., Saviano, S., and Piermarocchi, S. Carotenoids and antioxidants in age-related maculopathy italian study: multifocal electroretinogrammodifications after 1 year. Ophthalmology. 2008;115(2):324-333.
    • Pashkow, J., Watumull, D. G., and Campbell, C. L. Astaxanthin: a novel potential treatment for oxidative stress and inflammation in cardiovascular disease. Am J Cardiol. 5-22-2008;101(10A):58D-68D.
    • Serebruany, , Malinin, A., Goodin, T., and Pashkow, F. The in vitro effects of Xancor, a synthetic astaxanthine derivative, on hemostatic biomarkers in aspirin-naive and aspirin-treated subjects with multiple risk factors for vascular disease. Am J Ther.

    2010;17(2):125-132.

    • Yamashita The effect of a dietary supplement containing astaxanthin on skin condition. Carotenoid Sci. 2006;10:91-95.
    • Yoshida, , Yanai, H., Ito, K., Tomono, Y., Koikeda, T., Tsukahara, H., and Tada, N. Administration of natural astaxanthin increases serumHDL-cholesterol and adiponectin in subjects with mild hyperlipidemia. Atherosclerosis. 2010;209(2):520-523.
    • Yuan, P., Peng, J., Yin, K., and Wang, J. H. Potential health-promoting effects of astaxanthin: a high-value carotenoid mostly from microalgae. Mol Nutr Food Res. 2011;55(1):150-165.
     
       

     

     

     
       

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     
       

     

     

     

     

     

     

     

     

     

     

     

     
       
  • Cosmetic benefits of astaxanthin on humans subjects*

    Kumi Tominaga, Nobuko Hongo, Mariko Karato and Eiji Yamashita*

    Fuji Chemical Industry Co. Ltd., Kamiichi, Toyama, Japan

     

     

    Two  human  clinical  studies  were  performed.   One   was an open-label non-controlled study involving 30 healthy female subjects for 8 weeks. Significant im- provements were observed  by  combining  6  mg  per  day oral supplementation and 2 ml (78.9 μM solution)  per day topical application of astaxanthin. Astaxanthin derived  from  the  microalgae,   Haematococcus   pluvia- lis showed improvements  in  skin  wrinkle  (crow’s  feet  at week-8), age spot size (cheek at week-8), elasticity (crow’s feet at week-8),  skin  texture  (cheek  at  week-  4), moisture content of corneocyte layer  (cheek  in  10  dry skin subjects at week-8) and corneocyte condition (cheek at week-8). It may suggest that astaxanthin derived  from  H. pluvialis  can  improve  skin  condition in all layers  such  as  corneocyte  layer,  epidermis,  ba-  sal layer and dermis by combining oral supplementa-  tion and topical treatment. Another was a randomized double-blind placebo controlled study involving 36 healthy male subjects for 6 weeks. Crow’s feet wrinkle and elasticity; and transepidermal water loss (TEWL) were  improved  after  6  mg  of  astaxanthin  (the  same  as former study) daily supplementation. Moisture con- tent and sebum oil level at the cheek zone showed strong tendencies for improvement. These results sug- gest that astaxanthin derived from Haematococcus pluvialis may improve the skin condition in not only in women but  also in  men.

    Key words: astaxanthin, healthy human subjects, Haematococcus pluvialis, skin condition

    Received: 17 October, 2011; accepted: 01 March, 2012;

    available on-line: 17 March, 2012

     

     

    InTROduCTIOn

    Astaxanthin, is widely and naturally distributed in marine organisms, including crustaceans such as shrimps and crabs; and fish such as salmon and sea bream. In   fact, it is one of the oldest carotenoids isolated and iden- tified from lobster, Astacus gammarus (Kuhn et al., 1938). Astaxanthin was first commercially used for pigmen-  tation only in the aquaculture industry. Later in 1991, when the biological activity from potent antioxidative properties and the physiological function as a vitamin A precursor in fish and mammals (rats) were reported, asta- xanthin as a food supplement started gaining acceptance (Miki, 1991; Matsuno, 1991). Further reports suggest that astaxanthin does not have any pro-oxidative nature like β-carotene and lycopene (Martin et al., 1999) and its po- tent anti-oxidative property is exhibited on the cell mem- brane (Goto et al., 2001). Among various health-promot- ing effects of astaxanthin have been reported (Yuan et    al., 2010) including anti-inflammatory effects (Lee et al., 2003).  There  are  few  studies  on  the  skin.  In  terms of

     

    dermatological actions, suppression of hyper-pigmenta- tion (Yamashita, 1995) and inhibitions of melanin syn- thesis and photoaging (Arakane, 2002) have been report- ed. We have previously reported three clinical studies to evaluate either topical or oral supplementation effects of astaxanthin derived from the microalgae H. pluvialis. The first as a small pilot study was the repeated topical ap- plication test of a cream containing not only astaxanthin but other active ingredients  and  effective  base  materi- als (Seki et al., 2001). A double blind placebo controlled study using a dietary supplement containing astaxanthin and tocotrienol from palm oil was the second (Yamashi- ta, 2002). In the third study, we reported the effects of a dietary supplement containing only astaxanthin in a sin-  gle blind placebo controlled study (Yamashita, 2006). All these studies were either oral supplementation or topical application trials using female subjects. Here we report    an open-labeled non-controlled clinical study by combin- ing both oral supplementation and topical treatment of astaxanthin involving healthy female subjects and a ran- domized double-blind placebo controlled study by asta- xanthin oral supplementation involving 36 healthy male subjects.

     

    METhOd

    Materials. The material for oral supplementation con- tained AstaREAL® Oil 50F (Fuji Chemical Industry Co., Ltd., Toyama, Japan), 5% w/w astaxanthin H. pluvialis extract and canola oil as soft gel capsules. Each capsule contained 3 mg of astaxanthin. Identical placebo cap-  sules for control were prepared with only canola oil in  soft capsules.

    The product for external use had 0.094% AstaTROL™-Hp (5% w/w astaxanthin H. pluvialis ex- tract, Fuji Chemical Industry Co., Ltd., Toyama, Japan) resulting in 78.9 µM astaxanthin solution without any  other active ingredient and effective base   materials.

    Subjects and study design. As far  as  the  first  study (Study-1) is  concerned, thirty (30) healthy wom-  en in Japan, aged 20 to 55 years old, participated after obtaining their informed consent. Astaxanthin  wash- out period was  eight weeks  before start.  One  capsule as internal supplementation was administered to each subject twice daily, after breakfast and dinner respec- tively. 1ml of the topical application was applied onto the whole face of each subject twice daily every morn- ing and evening after washing. Test duration was eight weeks   starting   from   October,   2008. Measurements

     

     
       

     

    *e-mail: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.

    *Presented at the 16th International Symposium on Carotenoids, 17–22 July, 2011, Kraków, Poland

    Abbreviations: TWEL, transepidermal water  loss.

     

    of each test item were  performed  at  three  points,  at the beginning of the  study,  after  four  weeks  and  af- ter eight weeks. The study was open-label and non- controlled.

    On another study (Study-2), thirty-six (36) healthy  men in Japan, aged 20 to 60 years old, participated af-     ter obtaining their informed consent. The subjects were divided into the two groups, astaxanthin supplemented (n=18) and placebo (n=18). After an eight week wash-  out period one capsule was orally administered to each subject twice daily, after breakfast and dinner respec- tively. The test period was six weeks starting from October, 2008. Measurements of each test item were performed at the beginning of the study and after six weeks. The study was performed under a randomized double-blind  and  placebo  controlled manner.

    Conditions of measurement. The  measurements were performed 15 minutes after the subjects were al- lowed to rest in a seated position after washing their fac- es in an environmental test room conditioned to 20±2°C (or 68±3.6°F) room temperature and 45±10% relative humidity in the both  studies.

    Measurement parameters. Wrinkle. Skin surface photographs for crow’s feet condition evaluation were recorded using the Facial Stage (Moritex Co., Tokyo, Japan). Wrinkle topography measurements were made from negative skin replicas of the left crow’s feet and calculated by the ASA-03RXD (Asahibiomed Co., Ltd., Yokohama, Japan) image analysis based on six parame- ters — deepest point of the deepest wrinkle, mean depth of the deepest wrinkle, maximum  width  of  the  deep-  est wrinkle, area ratio of all wrinkles, mean depth of all wrinkles and volume ratio of all   wrinkles.

    Elasticity. Skin elasticity of the left crow’s feet area  was measured by ASA-GP1 (Asahibiomed Co.,   Ltd.).

    Age spot. Skin surface photographs for cheek condi- tion evaluation were also recorded using the Facial Stage with normal and UV lamps. Comparison of the most outstanding  age  spot  in  the  left  cheek  between  0  and

     

     

    Figure 1. Skin surface photographs and replica images of crow’s feet.

     

    8 weeks were determined by image analysis  using  Im- ageJ (NIH, USA).

    Skin texture. Skin topography for cheek (left side) condition evaluation were determined with  replicas  by the ASA-03RXD image analysis based on four  param- eters — number of texture, mean depth of texture,  volume ratio of texture/volume ratio of all texture and projection number of texture. Corneocyte of the left  cheek was collected by Scotch tape stripping and was applied to hematoxylin-eosin stain. The area was calcu- lated by ImageJ analysis on a prepared slide at a magni- fication  of  200 times.

    Moisture content. Skin moisture contents of the left crow’s feet for wrinkle evaluation and cheek for skin texture evaluation, respectively, were recorded using the ASA-M2 (Asahibiomed Co.,  Ltd.).

    Sebum oil: Skin sebum oil content at the left cheek  was measured by the SEBU sheet around the    nose.

    Transepidermal Water Loss (TEWL): TEWL at the left cheek was measured by the ASA-CT1 (Asahibiomed Co., Ltd.).

    Wrinkle, elasticity, age spots, skin texture  and  mois- ture  content  were  measured  for  Study-1.  And  wrinkle,

     

     

    Figure 2. Wrinkle parameters from replica image   analysis.

    By paired t-test: *p<0.05  **p<0.01.

     

    elasticity, moisture content, sebum oil and TWEL were measured for Study-2.

     

    RESuLTS

     

    Study-1

    Wrinkle, moisture content & elasticity. Figure 1 shows skin surface photographs and  replica  images  of the crow’s feet area in two subjects at week-0 and -8. Visual wrinkle reductions were observed respectively. Significant improvements on four parameters were ob- served. Deepest point of the deepest wrinkle (at week-      8 with p<0.01), mean depth of the deepest wrinkle (at week-4 and -8 with p<0.01), maximum width of the deepest wrinkle (at week-8 with p<0.01) and mean depth of all wrinkles (at week-4  and  -8  with  p<0.05)  out  of six as shown in Fig. 2. Moisture content of corneocyte layer in the left crow’s feet did not show any significant differences before and after the treatment (not shown). Elasticity of crow’s feet area significantly improved at  both week-4 and -8 (Fig.  3).

    Age spot. Figure 4 shows skin surface photographs with both normal and UV lamps of the cheek area in    two subjects at week-0 and -8. Visual age spot reduc-  tions were observed in the both subjects. The age spot area was significantly treated  at  week-8  as  shown  in  Fig. 5.

    Skin texture & moisture content. Figure 6  shows skin topographic replica images of the cheek in two sub- jects at week-8. Visual rough skin improvements were observed in the both subjects. There was a significant improvement on the parameter, mean depth of texture   (at week-8 with p<0.01) out of four as shown in Fig. 7. Total area of the corneocyte at week-8 significantly im- proved from the start period (Fig. 8). Moisture content

     

     
       

     

    Figure 3. Skin elasticity of crow’s   feet.

    By paired t-test: *p<0.05  **p<0.01.

     
       

     

     

    Figure 4. Skin surface photographs with both normal and uV lamps of cheek.

     

    Figure 5. Age spot area of   cheek.

    By paired t-test:  **p<0.01

     
       

     

    Figure 6. Skin topographic replica images of   cheek.

     
       

     

    Figure 7. Mean depth of texture of   cheek.

    By paired t-test: *p<0.05  **p<0.01.

     
       

     

    Figure 8. Size of total area of   corneocyte.

    By paired t-test:  *p<0.05.

    of corneocyte layer in the cheek among all subjects did  not show any significant differences.  However,  in  ten  dry skin subjects out of thirty showed a significant in- crease with p<0.05 at week-8 (not   shown).

    Study-2

    Wrinkle. As shown in Fig. 9, significant improve- ments in two parameters “Area ratio of all wrinkles”   and

     

     

    Figure 9. Wrinkle test parameters from replica image    analysis.

    Unpaired t-test: *p<0.05; N.S., not  significant.

     

                                                              

     

    Figure 10. Skin elasticity of crow’s   feet.

    Unpaired t-test: *p<0.05.

    “Volume ratio of all wrinkles” out of six parameters at week-6 were observed compared to the   start.

    Moisture content. Moisture content at  the  crow’s  feet did not show any significant differences before and after the administration (not shown). However, the mois- ture content of the cheek did show a tendency (p=0.08) increase among the selected subjects who had dry  skin less than 17 µS of moisture content at the beginning of  the study (data not  shown).

     
       

     

    Figure 11. Sebum oil of  cheek.

    Unpaired t-test: N.S., not  significant.

     

    Figure 12. Transepidermal water loss (TEWL) of   cheek.

    Unpaired t-test: **p<0.01.

    Elasticity. Elasticity of crow’s feet area significantly improved at week-6 compared to the start (Fig.    10).

    Sebum oil. Sebum oil of the cheek showed a tenden-  cy (p=0.085) of decrease at week-6 compared to the start (Fig. 11).

    TEWL. Figure 12 shows  a  significant  improvement on TEWL at week-6 compared to   week-0.

     

    dISCuSSIOn

    We studied the cosmetic effects of  astaxanthin,  a strong carotenoid antioxidant, from the two viewpoints   of administration technique and sex. Study-1 was per- formed to evaluate the impact of combination of both  oral supplementation and topical administration in an open-label non-controlled test involving female subjects. Significant improvements as a deep impact were ob- served in skin wrinkle (crow’s feet at week-8), age spot  size (cheek at week-8), elasticity (crow’s feet at week-8), skin texture (cheek at week-4), moisture content of cor- neocyte layer (cheek in 10 dry skin subjects at  week-8)  and corneocyte condition (cheek at week-8).  Combina- tion technique may be much beneficial for the skin. And Study-2 was performed to evaluate the efficacy in male subjects  by  oral  supplementation  under  a   randomized

     

    double-blind placebo controlled condition. Significant improvements were observed in wrinkle and elasticity of crow’s feet and TWEL at cheek at week-6 compared to start. Tendencies of improvement in moisture  content and sebum oil at cheek were also observed. Astaxanthin supplementation exhibited cosmetic benefits in not only female but male  subjects.

    The wrinkle parameters used in  the  present  studies  has been authorized by the Japanese Cosmetic Science Society (Task Force Committee for Evaluation of Anti- aging Function, 2007) as a guideline for the functional assessment of anti-wrinkle product. The Committee pre- scribes that a product can be effective in wrinkle reduc- tion in the case of significant improvement from at least one parameter. It is also provided that any parameters    are equivalent. Significant improvements from four pa- rameters in Study-1 and from two parameters in Study-2 were observed respectively. The female subjects  in Study-1 were unrestrained in any cosmetic behavior such as skin care or dietary supplement. Bedside, the male subjects in Study-2 were absent from any cosmetic be- havior. It seems that a double administration by combin- ing oral supplementation and topical treatment should be recommended for wrinkle reduction and oral supplemen- tation might be more potent than topical treatment. The mechanism of action of wrinkle reduction by astaxanthin could be explained as a dermis condition improvement through collagen fiber recovery. Astaxanthin promotes collagen fiber recovery by protecting the dermal layer  from singlet oxygen damage which has been substanti- ated by an in vitro study using human dermal fibroblasts (Tominaga et al., 2009). There are the reasons why the moisture content of corneocyte layer in crow’s feet was  not significantly changed before and after the test and    the ASA-GP is available to a measurement of dermic elasticity. Elasticity was also improved as a result of col- lagen fiber recovery both in Study-1 & -2. Significant in- hibition of melanogenesis in age spots were observed in the same manner as the other reports (Yamashita, 1995; Arakane, 2002) by suppressing the oxidative polymeriza- tion in melanocytes and inflammation in epidermis in Study-1. Regarding improvement of rough skin by asta- xanthin treatment on the mean depth of texture and the size of total area of the corneocyte in Study-1, it’s the    first finding. It seems that the improvements resulted in the moisture content increase in the cheek among dry  skin subjects. The mean moisture content in the all sub- jects at start was approximately 20 µS in Study-1. In gen- eral, the range of moisture content of dry skin is 12–15

    µS. A significant increase was observed in ten subjects whose moisture content was less than 17 µS at the start.  In Study-2 moisture content of the cheek show a strong tendency increase  among  the selected subjects less   than

    17 µS at the start. Topical treatment might be  more  deeply involved in the improvement of rough skin than oral supplementation. Corneocyte consists of the dead epidermal  cells.  Astaxanthin  treatment  might  normalize

     

    the corneocyte conditions protecting the keratinocyte differentiation and cornification from oxidative  dam-  ages such as inflammation in epidermis. Excess oxidized sebum oil causes rough skin and aging odor. It’s well- known that men have more sebum oil production than women. Astaxanthin supplementation may help to re-  duce rough skin and aging odor protecting the sebum     oil from peroxidation. TEWL is a marker for the bar-    rier functions in corneocyte layer. It also seems that the significant TWEL improvements resulted in normalizing the corneocyte condition. Atopic skin patients who have high TEWL may be treated by astaxanthin supplementa- tion.

    In conclusion these results may suggest that astaxan- thin derived from H. pluvialis can improve skin condition in all layers such as corneocyte layer,  epidermis,  basal layer and dermis by combining both oral  supplementa- tion and topical treatment and oral supplementation of astaxanthin can improve the skin condition in not only women but also  men.

     

    REFEREnCES

    Arakane K (2002) Superior skin protection via astaxanthin. Carotenoid Science 5: 21–24.

    Goto S, Kogure K, Abe K, Kimata K, Kitahama K, Yamashita  E, Terada H (2001) Efficient radical trapping at the surface and inside the phospholipid membrane is responsible for highly potent anti- oxidative activity of the carotenoid astaxanthin. Biochim Biophys Acta 1515: 251258.

    Kuhn R, Sorensen NA (1938) The coloring matters of the lobster (Astacus gammarus L.). Z Angew Chem 51:   465–466.

    Lee SJ, Bai SK, Lee KS, Namkoong S, Na HJ, Ha KS, Han JA, Yim    SV, Chang K, Kwon YG, Lee SK, Kim YM (2003) Astaxanthin inhibits nitric oxide production and inflammatory gene expression    by  suppressing  IkB  kinase-dependent  NF-κB  activation.  Mol  Cells

    16: 97–105.

    Martin HD, Ruck C, Schmidt M, Sell S, Beutner S, Mayer B, Walsh R (1999) Chemistry of carotenoid oxidation and free radical reactions. Pure Appl Chem 71:  2253–2262.

    Matsuno T (1991) Xanthophylls as precursors of retinoids. Pure Appl Chem 63: 81–88.

    Miki W (1991) Biological functions and activities of animal   carotenoids.

    Pure Appl Chem 63:  141–146.

    Seki T, Sueki H, Kono H, Suganuma K, Yamashita E (2001) Effects      of astaxanthin from Haematococcus pluvialis on human skin-patch test; skin repeated application test; effect on wrinkle reduction. Fragrance     J 12: 98–103.

    Task Force Committee for Evaluation of Anti-aging Function (2007) Guideline for evaluation of anti-wrinkle products in “Guidelines for evaluation of cosmetic functions”. J Jpn Cosmet Sci Soc 31: 411–431.

    Tominaga K, Hongo N, Karato M, Yamashita E (2009) Protective ef- fects of astaxanthin against singlet oxygen induced damage in hu-  man dermal fibroblasts in vitro. Food Style 21 13:    84–86.

    Yamashita E (1995) Suppression of post-UVB hyperpigmentation by topical astaxanthin from krill. Fragrance J 14:   180–185.

    Yamashita E (2002) Cosmetic benefit of dietary supplements includ-    ing astaxanthin and tocotrienol on human skin. Food Style 21 6: 112–117.

    Yamashita E (2006) The effects of a dietary supplement containing astaxanthin on skin condition. Carotenoid Science 10:   91–95.

    Yuan JP, Peng J, Yin K, Wang JH (2010) Potentialhealth-promoting effects of astaxanthin: a high-value carotenoid mostly from microal- gae. Mol Nutr Food Res 54:   1

  • Astaxanthin decreased oxidative stress and inflammation and enhanced immune response in humans

     

    Introduction

    Studies have reported important functions played by natural carotenoids in regulating immunity and disease etiology [1,2]. Specifically, interest in the biological activity of astaxanthin, an oxycarotenoid found in high amounts in the carapace of crustaceans and in the flesh of salmon and trout, has increased in recent years. In vitro studies have demonstrated that astaxanthin is sev- eral fold more active as a free radical antioxidant than b-carotene and a-tocopherol [3].

    Using a rodent model, we [4] and others [5,6] have demonstrated that astaxanthin stimulated immune response in mice. Mice supplemented with astaxanthin

     

    had increased ex vivo splenocyte antibody response to T-dependent antigens [6], lymphoblastogenic response and cytotoxic activity [4]. Moreover, these studies also showed that astaxanthin was consistently more active than other carotenoids such as b-carotene, lutein and canthaxanthin.

    In addition to immunoregulatory activity, astaxanthin also inhibited mammary tumor growth. We [7] reported that dietary astaxanthin inhibited mammary tumor growth in mice. Astaxanthin has been shown to reduce bacterial load and gastric inflammation in Helicobacter pylori-infected mice [5], and to protect against UVA- induced oxidative stress [8].

    Immune cells are particularly sensitive to oxidative

     

                                                                                                                  stress due to a high percentage of polyunsaturated fatty

     

    * Correspondence: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.

    1School of Food Science, Washington State University, Pullman, WA 99164- 6376 USA

     

    acids in their plasma membranes, and they generally produce more oxidative products [1]. Overproduction of

     

     
       

     

     

    © 2010 Park et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License  (http://creativecommons.org/licenses/by/2.0),  which  permits  unrestricted  use,  distribution,  and  reproduction  in any  medium, provided the  original work is properly   cited.

     

     

     

     

     

    reactive oxygen and nitrogen species can tip the oxidant: antioxidant balance, resulting in the destruction of cell membranes, proteins and DNA. Therefore, under condi- tions of increased oxidative stress (e.g. during disease states), dietary antioxidants become critical in maintain- ing a desirable oxidant:antioxidant balance. While stu- dies on the immunomodulatory role of dietary astaxanthin have been reported in rodents, similar stu- dies in humans are not available. We hypothesize that dietary astaxanthin will act as a potent antioxidative and anti-inflammatory agent; through these and other mechanisms, astaxanthin can enhance immune response. Our objective is to study the possible immune-enhancing, antioxidative and anti-inflammatory activity of dietary astaxanthin in humans.

     

    Subjects and methods

    Study participants and study  design

    Free-living healthy female college students with an aver- age age of 21.5 yr (20.2-22.8 yr) and BMI of 21.6 (16.3- 27.5) were participants in this study. Participants were recruited from Inha University (Seoul, Korea) through flyers and emails, and all were native Koreans. Subjects with a history of diabetes, alcohol abuse, cancer or smoking were excluded; exclusion criteria also included those taking antioxidant supplements. Prior to the initia- tion of dietary supplementation, a three-day dietary record was obtained from each subject who provided informed consent. During the study, subjects were allowed to consume their normal diets but were advised to refrain from eating astaxanthin-rich foods such as sal- mon, lobster, and shrimp. Subjects were ranked based on BMI (age was within a very narrow range) and groups of 3 participants with similar BNI were randomly assigned to receive daily: 0 (control; Con), 2 mg (2Asta), or 8 mg (8Asta) astaxanthin (109 g astaxanthin com- plex/kg oleoresin concentrate from Haematococcus plu- vialis, astaZanthin™, La Haye Laboratories Inc., Redmond, WA) (n = 14 subjects/diet) for 8 wk in a double-blind, placebo-controlled study. Astaxanthin was administered as a softgel capsule taken every morning, and all softgel capsules were externally identical. Blind- ing was further ensured by assigning consecutive num- bers to the dietary treatments and maintaining a master list until the study was completed. The astaxanthin complex used in this study came from a supercritical CO2 extract of Haematococcus pluvialis. Astaxanthin in the H. pluvialis extract is entirelythe 3S, 3S’ enantiomer, and is primarily monoesterified with smaller quantities of diester and free astaxanthin. The astaxanthin complex also contains small amounts (<15%) of mixed carote- noids including lutein, b-carotene and canthaxanthin. To minimize subject-to-subject and assay-to-assay varia- tion due to different sampling days, blood was drawn

     

    from all 42 subjects on one day for each of wk 0, 4 and

    1. Immune function and oxidative status was assessed within 24 h of blood collection. All procedures were approved by the Institutional Review Board (IRB #4421) of Washington State University.

     

    Analytical procedures

    HPLC

    Astaxanthin content in plasma was analyzed by reverse phase HPLC (Alliance 2690, Waters, Milford, MA) as pre- viously described [9]. Trans-b-apo-8’carotenal (Sigma Chem. Co., St. Louis, MO) was used as the internal stan- dard. Mobile phase used was acetonitrile:methanol:water, 47:47:16 (v/v/v), and samples were eluted through a 5-μm spherical C-18 column (3.9 × 150 mm Resolve, Waters, Milford, MA) at a flow rate of 1.5 mL/min. Absorbance was monitored at 492 nm on a photo diode array detector. Lymphoproliferation

    The proliferation response of peripheral blood mono- nuclear cells to phytohemagglutinin (2 and 10 mg/L final concentration), concanavalin A (2 and 10 mg/L), and pokeweed mitogen (1 and 5 mg/L) was assessed using whole blood cultures (to mimic in vivo conditions) as previously described [10]. Results were calculated as stimulation index.

    Natural killer cell cytotoxic  activity

    Effector cells (peripheral blood mononuclear cells) and target (K562) cells were cultured at effector:target ratios of 5:1 and 10:1 in DMEM (Sigma, St. Louis, MO) con- taining 100 mL/L fetal bovine serum, 0.1 U/L penicillin, and 100 g/L streptomycin sulfate. Killing was assessed using MTT to measure cell viability. The percent of spe- cific cytotoxicity was calculated as follows:

                                                           

     

    % Specific cytotoxicity   1   ODeffector target  ODeffector  / ODtarget   100.

     

    Phenotyping

    Populations of total T cells (CD3+CD19-), T cytotoxic cells (Tc; CD3+CD8+), T helper cells (Th; CD3+CD4+), B cells (CD3-CD19+), and natural killer cells (NK; CD3-/ CD16+56+) were quantitated by dual color flow cytome- try as previously described [10,11]. Cells were labeled with monoclonal antibodies conjugated to fluorescein isothiocyanate (FITC) or phycoerythrin (PE): anti-CD3 was conjugated to FITC, and anti-CD8, anti-CD4 and anti-CD19 were conjugated to PE (Caltag Laboratories, Burlingame, CA). In addition, the distribution of the intercellular adhesion molecule ICAM-1 (CD54+, BD Biosciences), and the leukocyte function antigens LFA-1 (CD11a+, BD Biosciences) and LFA-3 (CD58+, BD Bios- ciences) were measured. A lymphocyte analysis gate and the antibodies CD45-FITC and CD14-PE (Caltag Laboratories, Burlingame, CA) were used to help

     

     

     

     

     

    distinguish the lymphocytes from other blood cell types. A total of 2000 gated events were acquired for each sample and analyzed by flow cytometry (FACScan, BD Biosciences, San Jose, CA) using the Cell Quest program (version 3.3).

    Tuberculin delayed-type hypersensitivity

    Delayed-type hypersensitivity (DTH) response to an intracutaneous injection of tuberculin (Mono-Vacc Test O.T., Pasteur Merieux Connaught, France) was assessed on wk 8. A physician administered the injections and also measured skin thickness and induration at 0, 24, 48 and 72 h after challenge.

    Cytokine production

    Plasma samples were analyzed using commercially avail- able ELISA kits for IL-2 (BD OptEIA™ Set Human IL-2, BD Biosciences, San Diego, CA), TNFa  (BD OptEIA™ Set Human TNF), and IFN-g  (BD OptEIA™ Set Human IFN-g), as well as IL-1b (Amersham Pharmacia Biotech Inc., Piscataway, NJ) and IL-6 (Amersham Pharmacia Biotech Inc.).

    C-Reactive protein

    C-Reactive protein (CRP), a well-established marker of inflammatory status, was measured in plasma with a commercially available ELISA (Alpha Diagnostic, San Antonio, TX).

     

     

    Oxidative damage to  DNA

    Oxidative DNA damage was assessed by measuring plasma 8-hydroxy-2’-deoxyguanosine (8-OHdG) using competitive ELISA (BIOXYTECH® 8-OHdG-EIA Kit, OxisResearch, Portland, OR).

    Lipid-peroxidation

    Plasma concentrations of 8-epi-prostaglandin F2a (8-isoprostane) were measured by a commercially avail- able competitive ELISA (8-Isoprostane EIA kit, Cayman Chemical Company, Ann Arbor, MI).

     

    Statistical analysis

    Data were analyzed by repeated measures ANOVA using the General Linear Model of SAS [12]. Differences among treatment means were compared by a protected LSD test and considered different at P < 0.05.

     

    Results

    Plasma astaxanthin

    Astaxanthin was not detectable in the plasma of any subjects at wk 0 or in the conrol group at wk 4 or 8. However, concentrations of astaxanthin in plasma increased to maximal concentrations by wk 4 in a dose- dependent manner (Figure 1). Dietary recall showed no treatment difference in daily dietary intake (Table 1).

     

     

     

     

     

     

     

     

    Table 1 Composition of averaged 3-day dietary recall within subject treatment groups (n = 14/diet treatment group) prior to daily supplementation with 0, 2 or 8 mg astaxanthin

     

    Astaxanthin

     

    0 mg

    2 mg

    8 mg

    Total Kcal (Kcal/d)

    1777

    1618

    1736

    Protein (g/d)

    64

    59

    68

    Fat (g/d)

    54

    49

    52

    CHO (g/d)

    258

    238

    254

    a-Carotene (μg/d)

    457

    527

    567

    b-Carotene (μg/d)

    1813

    1664

    1773

    b-Cryptoxanthin (μg/d)

    61

    81

    38

    Lutein & Zeaxanthin (μg/d)

    610

    490

    338

    Lycopene (μg/d)

    2067

    2327

    1133

    There were no differences between groups, as determined by T test. Values are  means of 3-day dietary  recalls.

     

     

    Lymphoproliferation

    Proliferation of peripheral blood mononuclear cells was consistently higher (P < 0.05) when stimulated with both T cell-dependent (phytohemmaglutinin, concanava- lin A) and B cell-dependent (pokeweed mitogen) mito- gens in 8Asta on wk 8 (Figure 2). Both concentrations of each mitogen showed similar trends whether mito- gens were low or high concentration. No differences in response were observed in 2Asta.

     

    NK Cytotoxicity

    Higher (P < 0.05) NK cell cytotoxic activity (effector:tar- get cell ratio of 10:1) was seen in 8Asta but not in 2Asta by wk 8 (Table 2).

     

     

    Phenotyping

    The population of CD3+ total T cells was higher (P < 0.05) in 2Asta or 8Asta compared to Con on both wk 4 and 8 (Table 2). The percent B cell population was higher (P < 0.05) only in 2Asta on wk 8 (Table 2). On the other hand, dietary astaxanthin did not significantly influence the population of Th, Tc or NK cells or the ratio of Th:Tc cells (Table 2).

    On wk 8, the frequency of cells expressing CD11a+ LFA-1 marker was higher in 2Asta (42%) but not those given 8Asta (30.6%) compared to Con (32%). Supple- mental astaxanthin did not have a significant effect on the expression of the cell surface adhesion molecules ICAM-1 (CD54) and LFA-3 (CD58) (data not shown).

     

    Tuberculin DTH test

    DTH response was maximal at 48 to 72 h post-chal- lenge (Figure 3). Subjects fed 2Asta had higher (P < 0.05) DTH response than Con, whereas 8Asta did not show a similar DTH response.

     

     

     

     

     

     

     

     

    Cytokines

    No differences in TNF-a and IL-2 levels were seen in any treatments (Table 3). By wk 8, IFN-g concentrations were higher in 8Asta (P < 0.05) (Table 3); IFN-g in 2Asta and Con did not change throughout the study. Treatment 8Asta also showed an increase (P < 0.05) in IL-6 on wk 8.

     

    C-Reactive  protein

    The concentration of plasma C-reactive proteins was lower (P < 0.05) in 2Asta on wk 8 compared to the Con (Figure 4). However, higher dietary astaxanthin amounts

     

     

     

     

     

    Table 2 Immune cell response following daily supplementation with 0, 2 or 8 mg astaxanthin   after

    0, 4 and 8  wk

    Treatment                            Wk 0            Wk 4            Wk 8

    Total T cells (%)

    0 mg                               65.9 ± 2.1     64.4 ± 1.9b      70.6 ±  1.5b

    2 mg                               69.5 ± 1.4     69.6 ± 1.5a      75.7 ±  1.6a

    8 mg                               67.2 ± 2.2      69.6 ± 1.8a     74.3 ± 1.8ab

    Total Th  cells (%)

    0 mg                               35.3 ± 1.5     36.3 ± 1.7     39.6 ± 1.4

    2 mg                               35.3 ± 1.8     36.0 ± 1.9     39.7 ± 2.0

    8 mg                               35.2 ± 1.5     34.3 ± 1.4     38.4 ± 1.2

    Total Tc cells (%)

    0 mg                               26.7 ± 1.4     26.1 ± 1.2     28.4 ± 1.4

    2 mg                               28.2 ± 1.9     27.6 ± 1.9     28.7 ± 1.6

    8 mg                               28.4 ± 1.6     28.3 ± 1.6     28.8 ± 2.1

    Th:Tc  ratio cells

    0 mg                               1.35 ± 0.07    1.43 ± 0.09    1.48 ± 0.10

    2 mg                               1.31 ± 0.15    1.35 ± 0.16    1.46 ± 0.15

    8 mg                               1.31 ± 0.13    1.29 ± 0.13    1.37 ± 0.14

    Total B cells (%)

    0 mg                               11.4 ± 0.9     11.1 ± 0.5     10.7 ± 0.5b

    2 mg                               13.1 ± 1.1     12.7 ± 1.2     13.1 ± 0.5a

    8 mg                               13.3 ± 1.4     11.9 ± 0.7     11.1 ± 0.3ab

    Total NK cells (%)

    0 mg                               13.7 ± 0.9     12.6 ± 1.2     10.6 ± 0.8

    2 mg                               12.5 ± 1.0     13.1 ± 1.9     12.6 ± 1.5

    8 mg                               13.4 ± 1.1     15.6 ± 1.4     13.1 ± 1.4

    NK  cytotoxic activity(% lysis)

    0 mg                               51.5 ± 4.4     51.0 ± 2.5     57.8 ± 2.7b

    2 mg                               58.2 ± 3.8     54.4 ± 2.9     57.1 ± 2.6b

    8 mg                               56.1 ± 3.1     54.3 ± 1.9     67.9 ± 3.0a

    a, bDifferent letters represent significant treatment differences (P < 0.05) as analyzed  by  ANOVA. Values  are means  ± SEM.

     

    did not influence the concentration of this acute phase protein.

     

    DNA Damage

    Concentrations of 8-OHdG were dramatically lower (P < 0.01) as early as wk 4 in 2Asta and 8Asta (Figure 5). DNA damage observed with 2Asta was not further decreased in the group fed higher dietary astaxanthin (8Asta).

     

    Lipid Peroxidation

    Dietary astaxanthin did not significantly influence con- centrations of plasma 8-isoprostane at all periods stu- died. The overall mean concentration of 8-isoprostane was 30.8 ± 0.4 pg/mL across all treatments.

     

    Discussion

    While the biological action of astaxanthin has been reported in both in vitro and in vivo studies, these have mainly used rodents and in vitro models. This is the first comprehensive study to examine the action of diet- ary astaxanthin in regulating immune response, oxida- tive damage and inflammation in humans. Dietary astaxanthin enhanced both cell-mediated and humoral immune responses in young healthy feamles. The immune markers significantly enhanced by feeding astaxanthin included T cell and B cell mitogen-induced lymphocyte proliferation, NK cell cytotoxic activity, IFN-g and IL-6 production, and LFA-1 expression. Enhancement of these ex vivo immune markers corre- sponded with increased number of circulating total T and B cells. In addition, subjects given astaxanthin also showed an enhanced tuberculin DTH response, a reli- able clinical test to assess in vivo T cell function. All of


     

     

     

     

    Table 3 Cytokine response following daily supplementation with 0, 2 or 8 mg astaxanthin   after

    0, 4 and 8  wk

     

    Treatment                                       Wk 0        Wk 4         Wk 8

     

    TNF-a (pg/mL); Overall SE = 0.43

    0 mg                                          1.14           1.63           1.43

    2 mg                                          0.80           1.13           1.44

    8 mg                                          1.51           2.15           2.60

    IFN-g (pg/mL); Overall SE = 0.34

    0 mg                                          5.85           4.47           4.68b

    2 mg                                          4.87           4.26           5.00b

    8 mg                                          4.67           6.23           9.55a

    IL-6 (pg/mL); Overall SE = 2.9

    0 mg                                          10.5           12.7           13.6b

    2 mg                                          10.0           11.5           8.7b

    8 mg                                          11.5           12.4           25.2a

    IL-2 (pg/mL); Overall SE = 0.10

    0 mg                                          8.61           8.04           7.74

    2 mg                                          5.09           4.67           4.46

    8 mg                                          4.91           4.01           3.90

     

    a, bDifferent letters represent significant treatment differences (P < 0.05) as analyzed by ANOVA. Values are means; variation is expressed as a representative overall standard  error.

     

     

    these immune responses were generally observed after 8 wk of supplementation following a cutaneous tuberculin injection.

    Modulatory actions of astaxanthin on immune response have been demonstrated in both in vitro and in vivo studies. We previously reported higher mitogen- induced splenocyte proliferation in mice [4], dogs [13] and cats [14] fed astaxanthin. Astaxanthin stimulated

     

     

    cell proliferation of murine splenocytes and thymocytes in vitro [15]. Others have shown that astaxanthin increased cytotoxic T lymphocyte activity in mice [16] and inhibited stress-induced suppression of NK cell activity [17]. In this study, astaxanthin heightened NK cell cytotoxic activity. Natural killer cells serve in an immuno-surveillance capacity against tumors and virus- infected cells; therefore, astaxanthin may play a role in cancer etiology. Patients with Chediak-Higashi syn- drome, a disorder associated with defective NK cell function, are indeed more susceptible to tumor formation.

    Flow cytometry data showed higher subpopulations of total T and B cells. Activated T cells and NK cells pro- duce IFN-g, which is involved in immune-regulation, B cell differentiation, and antiviral activity. IFN-g produc- tion was higher in subjects supplemented with astax- anthin, similar to the response in mice given astaxanthin [16]. Splenocytes of tumor-bearing mice fed lutein also had higher IFN-g expression, and these changes paralleled the inhibitory action of lutein against tumor growth [18]. Astaxanthin decreased bacterial load and gastric inflammation in mice infected with Helico- bacter pylori by shifting the T-lymphocyte response from a Th1 response dominated by IFN-g to a Th1/Th2 response dominated by IFN-g and IL-4 [5]. Modulation of the humoral immune response also occurs; astax- anthin increased antibody production in mouse spleno- cytes [19], partially restored humoral immune response in old mice [6], enhanced immunoglobulin production in response to T-dependent stimuli in human blood cells [20] and induced production of   polyclonal

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

     

    antibodies G and M in murine spleen cells [15]. The present study suggests that the higher antibody produc- tion may be due to an increase in B cell number.

    The skin tuberculin test is a reliable clinical test to assess in vivo T cell function. This study shows that subjects given astaxanthin had a heightened DTH response, which is also seen in dogs [13] and cats fed astaxanthin [14], b-carotene [10,14] and lutein [21,22]. Leukocyte function antigens (LFA) mediate intercellular adhesion between leukocytes and other cells in an anti- gen non-specific fashion. LFA-1 is a b2-integrin expressed on leukocytes involved in the migration of lymphocytes, monocytes and neutrophils. LFA-1 binds to ICAM-1 and ICAM-2 expressed on vascular endothe- lium, and controls lymphocyte migration into inflamma- tory sites. Endothelial expression of ICAM-1 is inducible while ICAM-2 is constitutive. Therefore, the heightened DTH response in this study is likely due to an increased expression of LFA-1 but not ICAM.

    Dietary astaxanthin dramatically decreased one DNA damage biomarker (plasma 8-OHdG), and this protec- tive effect was observed by wk 4 of feeding. Maximal

     

    response was observed with the lower 2 mg astaxanthin dose. In addition, subjects fed 2 mg astaxanthin also showed lower plasma C-reactive protein concentrations, demonstrating the anti-inflammatory action of astax- anthin in humans. Immune cells are particularly sensi- tive to membrane damage by free radicals. Reactive oxygen species (ROS) are produced via the mitochondria electron transport system during ATP production, xanthine oxidase and phagocytes [23,24]. In fact, cumu- lative oxidative damage to the mitochondria is consid- ered the main culprit of cell senescence which in turn is responsible for aging and the development of age-related chronic diseases [25]. The ROS can induce redox-sensi- tive transcription factors such as NFkB and AP-1, which regulate genes controlling production of chemokines, inflammatory cytokines, and adhesion molecules which stimulate phagocytic infiltration [26]. Conversely, astax- anthin, acting as a potent antioxidant, can inhibit ROS- induced production of these transcription factors, thereby decreasing inflammation. Indeed, astaxanthin attenuated exercise-induced neutrophil infiltration and subsequent delayed-onset damage to the gastronemius

     

     

     

     

     

    and heart muscle in mice [26]. Astaxanthin is reported to be approximately 100 fold more protective than lutein and b-carotene against UVA-induced oxidative stress in vitro [8].

    Reactive nitrogen species also play an important role in inflammation. As with ROS, astaxanthin has been reported to decrease the production of nitric oxide (NO) and iNOS activity in a mouse macrophage cell line, resulting in the inhibition of COX which down-reg- ulates the production of PGE2 and TNF-a [27]. TNF-a is a pleiotropic cytokine produced by activated macro- phages and monocytes, and has nonspecific resistance against various infectious agents. Similarly, Lee et al.

    [28] reported that astaxanthin suppressed serum NO, TNFa and IL-1b in mice injected with lipopolysacchar- ide. The TNF-a and IL-1 cascade activates p38 MAPK, thus promoting proinflammatory gene expression and cytokine production. Therefore, in this study, astax- anthin exerted its anti-inflammatory action by inhibiting reactive oxygen and nitrogen species.

    Why dietary astaxanthin did not reduce lipid peroxi- dation is unclear. Astaxanthin has been shown to be one of the most effective antioxidants against lipid per- oxidation and oxidative stress in in vitro and in vivo sys- tems [3,29]. Humans given 1.8 to 21.6 mg astaxanthin daily for 14 d increased the lag time for LDL oxidation [30]. Astaxanthin is as effective as a-tocopherol in inhi- biting radical-initiated lipid peroxidation in rat liver microsomes [31], and is 100 times more active than a-

    tocopherol in protecting the rat mitochondria against Fe2+-catalyzed lipid peroxidation in vivo and in vitro [3]. The potent antioxidant activity of astaxanthin is likely

    due to the presence of a keto- and a hydroxyl group on each end of its molecule. This structural property effec- tively rigidifies cell membranes, thereby limiting the penetration of lipoperoxidation promoters across the lipid bilayer [32]. The isoprostane methodology used in this study lacks sensitivity and accuracy; this may account for the lack of a significant effect seen in this study.

    Taken together, the immunomodulatory, antioxidative and anti-inflammatory activity of astaxanthin will likely influence the etiology of cancer and inflammatory diseases. Astaxanthin was more active than b-carotene, lutein and canthaxanthin in inhibiting mammary tumor growth in mice [7]. Others have reported that astaxanthin protected against carcinogenesis of the urinary bladder [33], decreased cancerous growth of the mouth [34], and decreased the number and size of liver preneoplastic foci

    [35] in rodents. Astaxanthin, as an algal extract, protected UVA-induced DNA damage in human skin fibroblasts (IBR-3), melanocytes (HEMAc) and intestinal CaCo2 cells [36]. In addition, astaxanthin ameliorated other oxidative stress-induced inflammatory diseases such as diabetic

     

     

    nephropathy in diabetic mice [37], lipopolysaccharide- induced uveitis in rats [27], and exercise-induced skeletal and cardiac muscle damage in mice [26].

    The polar ends if the astaxanthin structure allows it span biological membranes; this transmembrane align- ment allows astaxanthin to preserve the membrane structure [38], decrease membrane fluidity [39], and function as an antioxidant [40]. These and other mechanisms may explain the antioxidative, anti-inflam- matory and immune-modulatory action of astaxanthin.

    In this study, plasma concentrations of astaxanthin in subjects given 2 or 8 mg astaxanthin daily for 4 wk increased to 0.09 to 0.13 μmol/L, with no further increase observed at 8 wk. These plasma concentra- tions are lower than that reported by Osterlie et al.

    [41] who showed maximal concentrations of 1.3 mg/L (2.28 μmol/L) in subjects administered a single oral dose of 100 mg astaxanthin. The difference in plasma concentrations in the two human studies is expected, due to differences in the dose and length of astax- anthin administration; however, the two studies also used different sources of astaxanthin and different sub- ject gender and age. Most of the astaxanthin was found in the VLDL chylomicra, with lesser amounts in the LDL and HDL [41]. While the stereoisomer form of astaxanthin was not identified in this study, Osterlie et al. [41] reported a preferential uptake of the Z-iso- mers as compared to the all-E-astaxanthin. Astax- anthin used in the present study is from Haematococcus pluvialis and exists primarily in an esterified 3S, 3’S stereoisomer while synthetic astax- anthin [41] is primarily the 3R, 3’S form. The amount of supplemental astaxanthin used in this study is achievable through diet means. For instance, the astax- anthin content of salmon flesh ranges from 3 to 37 mg/kg [42,43]; therefore, a 200-g serving of salmon provides approximately 1 to 7 mg astaxanthin. Wild salmon contains the 3S, 3S’ form of astaxanthin almost exclusively. The 3R, 3R’ form is found rarely in nature but does exist in some crustaceans such as in Krill. In healthy humans, 6 mg astaxanthin from H. pluvialis algal extract can be safely consumed [44].

    Overall, this study shows that dietary astaxanthin enhanced immune response, and decreased a DNA oxi- dative damage biomarker and inflammation in young healthy females. It is the initial scope of the study to focus on a narrow population with regards to age, gen- der and race; however, antioxidants generally show greater physiologic modulation under excess amounts of oxidative stress, in immuno-compromised individuals, and with longer feeding periods. These likely explain the lack of efficacy in certain response measures studied. Future studies with astaxanthin administration will include these parameters. However, our present study

     

     

     

     

     

    suggests astaxanthin to be a bioactive natural carotenoid that may be important to human health.

     

    Acknowledgements

    This work was supported by a grant from the Washington Technology Center, Seattle WA and La Haye Labs, Inc., Redmond,   WA.

     

    Author details

    1School of Food Science, Washington State University, Pullman, WA 99164- 6376 USA. 2Food and Nutrition, Inha University, Incheon, Korea. 3La Haye Labs, Inc, Redmond, WA, USA.

     

    Authors’ contributions

    JSP and BPC designed research, analyzed data, and wrote the paper; JSP,  BPC, JHC, and YKKconducted research; LLL provided essential materials; BPC had primary  responsibility  for  final  content.  All  authors  read  and  approved the  final  manuscript.

     

    Competing interests

    The authors declare that they have no competing   interests.

     

    Received: 8 January 2010  Accepted: 5 March    2010

    Published: 5 March  2010

     

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  • Omega-3, Omega-6 and Omega-9 Fatty Acids: Implicationsfor Cardiovascular and Other Diseases

    Melissa Johnson1* and Chastity Bradford2

    1College of Agriculture, Environment and Nutrition Sciences, Tuskegee University, Tuskegee, Alabama, USA

    2Department of Biology, Tuskegee University, Tuskegee, Alabama, USA

     

      

     

     

     

    Keywords: Omega-3 fatty acids; Omega-6 fatty acids; Omega-9 fatty acids; Cardiovascular disease; Hypertension; Inflammation

    Introduction

    Strategic in pathophysiological  homeostasis  (following  injury),  as well as cellular, tissue and organismic protection are acute and chronic inflammatory responses [1,2]. Consequently, the pathogenesis and progression of cardiovascular and other diseases is initiated and perpetuated by this phenomenon. Efforts to normalize or control inflammatory processes include pharmacological, dietary and behavioral therapies, aimed at regulating biologically stimulatory molecules that may stimulate or suppress the synthesis of inflammatory triggers and subsequent byproducts [3-9]. The most recognizable potent bioactive lipid mediators are Arachidonic Acid (AA, C20:4n6), Eicosapentaenoic Acid (EPA, C22:5n3) and Docosahexaenoic Acid (DHA, C20:6n3), synthesized from their dietarly essential precursors linoleic (LA, C18:3n6) and α- linolenic (ALA, C18:3n3) acids (Figure 1). The omega-9 fatty acid, oleic acid, has been suggested to occupy     a role in the metabolism of the essential fatty acids [10,11]. These bioactive lipid mediators regulate pro-and anti-inflammatory processes via their ability to stimulate enzymes and produce cytokines and other acute phase molecules [12]. Further, these mediators occupy a central role in the synthesis of lipoxins and resolvins that hinder inflammatory pathways, increase the production of anti-inflammatory cytokines and facilitate the resolution of acute inflammation [13-17]. Decreasing dietary omega-6 fatty acid (i.e. linoleic acid) intake increases the bioavailability of omega-3 fatty acids [18], which may in turn lower

     

    genetic regulation and signaling, suggesting that these profiles may be useful clinical diagnostic and therapeutic tools [22-57]. This review provides a brief synopsis of the structure, function and physiological implications of the omega-3, omega-6 and omega-9 fatty acids in inflammation, hypertension, and Cardiovascular Disease (CVD).

    Omega fatty acids and inflammation

    Inflammation, resulting from various genetic, demographic, behavioral, environmental and nutritional interactions, is at the center of CVD and other  vascular  diseases  (Figure  2).  Potential  triggers  of increased risk for inflammation and subsequent endothelial and vascular injury are genetic characteristics [58], Western dietary patterns [59], environmental toxins [60], adaptive immune responses [61], the presence of other co-morbidities [62,63], and socioeconomic factors [64]. This is evident in the new paradigm shift of evaluation of heart failure patients with preserved ejection fraction. The emphasis shifts from solely using left ventricular afterload to evaluate heart failure patients, and now includes coronary microvascular inflammation [65] thus, changing the methods of patient evaluation. Omega fatty acids have been described as inflammation-modulating agents, which may stimulate or suppress the synthesis of pro- and/or anti-inflammatory cell signaling molecules. In a recent randomized controlled trial, omega-3 polyunsaturated fatty acid supplementation lowered the concentration of serum proinflammatory cytokines[66].

    One of the omega-6 fatty acids, arachidonic acid, directly impacts inflammation. Invitro it enhanced the ability of endothelial cells to bind

     

    tissue concentrations of the omega-6/omega-3 fatty acid ratio, mitigate                                                                                                                        the intensity and duration of inflammatory responses and subsequently

     

    reduce disease risk [19-21].

    The relationship between omega-3 and omega-6 fatty acids, inflammation and disease pathogenesis continues to be a topic of extensive study. To a lesser magnitude omega-9 fatty acids have been considered as potential disease mediators. These fatty acids may work individually, additively or synergistically as precursors and critical elements within metabolic pathways, thus actively influencing and/   or altering membrane fluidity, cell structure, and disease pathogenesis (Table 1). Research has revealed the relationship between inflammation and the cellular lipidomic (i.e. lipid) and glycomic (i.e. sugar)  profiles,

     

    *Corresponding author: Melissa Johnson, College of Agriculture,   Environment

    and Nutrition Sciences, Tuskegee University, Tuskegee, Alabama, USA, Tel: 334-727-8625; E-mail: Questo indirizzo email è protetto dagli spambots. È necessario abilitare JavaScript per vederlo.

    Received  September  02,  2014;  Accepted  September  26,  2014;   Published

    September 30, 2014

    Citation: Johnson M, Bradford C (2014) Omega-3, Omega-6 and Omega-9 Fatty Acids: Implications for Cardiovascular and Other Diseases. J Glycomics Lipidomics 4: 123. doi:10.4172/2153-0637.1000123

    Copyright: © 2014 Johnson M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

     

     

     

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    monocytes- thus, facilitating the pro-inflammatory process. Linoleic and γ-linolenic, omega-6 fatty acids, and omega-9 oleic acids were able to indirectly provoke the synthesis of Reactive Oxygen Species (ROS) superoxide, a pro-inflammatory mediator, mainly by activating p47 and NADPH oxidase enzyme complex [67]. Oleic acid also induced foam cell formation in rat aorta smooth muscle cells and enhanced atherosclerotic lesion development [68]. This is of particular interest as macrophage foam cell has been suggested to be a potential target  for therapeutic interventions [69], with the oxidative byproducts of cholesterol metabolism being found to influence the lipidome and transcriptome of the macrophage [70]. Others found the activation

     

    of macrophages to regulate the expression of genes involved in lipid metabolism, immunity and apoptosis [71,72].

    An alternative study found that oleic acid exerted vascular antiatherogenic effects [54] Oleic acid was able to mitigate the effects of TNF-α-induced oxidative stress and injury in adult male Sprague- Dawley rat cardiomyocytes [73] as well as reduce the inflammation associated with saturated fatty acid-induced inflammation in human aortic endothelial cells [74]. Further, the incorporation of milks enriched with oleic acid into the diet has resulted in reductions in total cholesterol, LDL-cholesterol and triglyceride levels, the effects of which

     

     

     

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    Table 1: Structure, dietary source, mechanism and implications of select omega-3, omega-6 and omega-9 fatty acids.

     

    to the endothelium.The endothelial cells are in direct contact with   the red blood cells, and blood lipid profiles are tools of evaluating cardiovascular health. Fatty acid composition of a major component of the endothelium, caveolae, played a regulatory role in TNF-α- induced endothelial cell activation and inflammation. The major omega-6 unsaturated fatty acids in the American diet are atherogenic and enhance the endothelial inflammatory response [76]. One of  them, arachidonic acid, directly impacted inflammation. In vitro, it enhanced the ability of endothelial cells to bind, a pro-inflammatory response [77]. In younger animals, estrogen inhibits the expression    of proinflammatory mediators in vascular smooth muscle cells [78]. Therefore, in addition to the fatty acids, age and gender play a major role in the inflammatory response. Further, a higher eicosapentaenoic acid to arachidonic acid ratio was associated with decreased LV wall thickness among individuals with diabetes [79]. The ability of omega-3,

     

    Figure 2: Interactions between and among factors contributing to  inflammatory

    status and disease risk.

     

    6, and 9 fatty acids to differentially modulate inflammatory stimuli,

     

                                                                                                                            impact  vascular  composition,  cellular  responsiveness,  and influence

     

    were observed among healthy individuals, those with increased risk for cardiovascular disease and individuals with CVD [75]. Although studied to a much lesser degree than oleic  acid,  another  omega-9 fatty acid, nervonic acid, has demonstrated influence on CVD risk. Researchers found Body Mass Index (BMI), leptin, triglycerides, total cholesterol and fasting blood glucose to be significantly negatively correlated with serum nervonic acid. These findings illustrate the ability of nervonic acid to exert protective effects against obesogenic- linked risk factors and conditions such as insulin resistance, diabetes, dyslipidemia and metabolic syndrome.

    The impact of fatty acids as inflammatory-modulators is crucial   to the state of the vasculature.The vasculature is mainly comprised    of endothelial cells, caveolae, smooth muscle cells, adventitia, and fibroblasts. Thus, the cellular responsiveness of the vasculature is  vital

     

    the structural integrity of the left ventricle underscore the implications of these fatty acids in chronic disease risk and prevention.

    Omega fatty acids and hypertension

    Chronic diseases such as hypertension, obesity, and diabetes are a national and international concern. Obesity prevalence has increased dramatically in recent years. The mortality of obese patients is more often a result of diabetes and hypertension [80]. Obesity is strongly associated with metabolic abnormalities, including insulin resistance, type 2 diabetes, hypertension, and dyslipidemia, mediated in part by the chronic inflammatory state induced by the secretion of adipocytokines, such as angiotensinogen, transforming growth factor–beta, tumor necrosis factor–alpha, and interleukin-six [81-83].

    The  cardioprotective  mechanisms  of  the  omega-3  fatty     acids

     

     

     

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    Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA)  have been attributed to their ability to displace the omega-6 fatty acid, arachidonic acid [8], as molecular substrates during the cyclooxygenase and oxygenase pathways. The combined hypotensive effects of EPA and DHA have been demonstrated in randomized controlled trials [84]. However others found DHA and DHA epoxides to be effective  in lowering blood pressure but not EPA [85]. The epoxides of an omega-6 fatty acid, arachidonic acid epoxyeicosatrienoic acids also exhibit antihypertensive and anti-inflammatory effects [86]. Actions of these fatty acids subsequently influence metabolism, β-oxidation, fatty acid synthesis, pro-inflammatory molecule synthesis and the transcription of genes coding for transcription factors (e.g. Peroxisome Proliferator-Activated Receptor [PPAR], Sterol-Response Element Binding Protein [SREBP] and Nuclear Factor jB [NF-jB] as well as enzymes implicated in cholesterol synthesis) [87,88]. Intake of EPA and DHA has been inversely associated with markers of inflammation in both men and women [89] In addition to influencing cytokine concentrations, EPA and DHA have been demonstrated to influence blood glucose and lipid profile [90]. The supplementation of DHA into

     

    the diet of hypertriglycemic men was found to decrease serum levels of c-reactive protein and other inflammatory biomarkers [91].

    Studies suggest that there is a role for the renin-angiotensin system in the mechanistic blood pressure lowering effects of omega-3 fatty acids.The Ren-2 rat model is mediated by ANG II, and the data suggest that omega-3 PUFA may reduce hypertension via the renin-angiotensin system [92]. In models of Angiotensin-II induced  hypertension,  DHA epoxides reduce inflammation and systolic blood pressure partially via reduction of prostaglandins, MCP-1, and upregulation of angiotensin-converting enzyme-2. It has been proposed that the oleic acid constituent of olive oil may be responsible for the hypotensive and cardio protective effect associated with olive oil consumption [93- 96]. Flaxseed, one of the richest sources of the plant-based omega-3 fatty acid, alpha-linolenic acid has been suggested to have a positive impact on CVD.There is strong scientific evidence from human trials that omega-3 fatty acids from fish or fish oil supplements (EPA and DHA) can significantly reduce  risk  factors  for  heart  disease  (such as reducing blood triglyceride [TG] levels, LDL-cholesterol, serum lipids, blood glucose), diabetes and metabolic syndrome [97-100],  yet

     

     

     

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    using nutritional strategies to combat diseases is not the first line of therapeutic intervention [101,102]. Unfortunately, analysis of national observational data indicates that U.S. adults are not consuming the recommended intake of fish and omega-3 fatty acids [103].

    Omega fatty acids and other diseases

    In addition to suppressing or inhibiting the expression of specific genes implicated in lipid metabolism, dietary fatty acid intake influences cellular, molecular oxidative and inflammatory status [8]. In addition to occupying a role in immune function [104], oleic acid inhibits food intake and glucose production in male rats [105] and has been suggested to enhance insulin production in rat pancreatic beta cells in both in vivo and in vitro environments favoring the inhibition of insulin production by TNF-α [106]. Further, the presence of a rich supply of oleic acid within low density lipoprotein molecules was protective against oxidative modification in rabbits, suggesting the antiatherogenic propensity of oleic acid. Conversely oleic acid was able to facilitate increased macrophage concentrations in mesenteric adipose tissue [107] and attenuate renal fibrosis [108]. Although  omega-3  fatty acids have been classified as anti-inflammatory mediators, there is conflicting evidence on the definite ability of these fatty acids to consistently reduce the risks, morbidities and mortalities associated with CVD, cancers and other inflammatory diseases and disorders [109]. There is also evidence for the role of omega-3 fatty acids in the stress response and cognitive function. Rats fed the omega-3 enriched diet had a lower stress-induced weight loss and plasma corticosterone peak, and reduced grooming [110]. These data suggest that the response to chronic restraint stress can also be altered by omega-3 fatty acids.

    Conclusions

    Central to  the  initiation,  pathogenesis  and  progression  of  many disease states is inflammation. Conventional mechanisms of alleviating inflammation include pharmacological therapies, which often target specific key components of inflammatory pathways.  Albeit not  relatively  novel,  increased  attention  has  been  devoted  to more aggressively reevaluating dietary approaches that mitigate inflammatory sequelae. Serving as mediators of lipid metabolism and foundational biomolecules of the lipidome, the character of omega-3, omega-6 and omega-9 fatty acids warrants further discussion. Omega-3 and omega-6 fatty acids have typically been associated with anti- and pro-inflammatory pathways, respectively, whereas the direct role of omega-9 fatty acids in inflammatory pathways remains unclear. In conjunction with other fatty acids and lipid classes, the omega-3, -6 and -9 fatty acids make up the lipidome, and within the conversion of excess carbohydrates into fats, transcendence of the glycome into the lipidome occurs.

    More recently, lipidomics profiling has been used as an assessment and monitoring tool for cardiovascular and other disease risk [23,111]. Bioinformatical tools have been particularly useful in examining the lipidome [112]. The genetic, metabolic and phenotypic consequences of omega-3, omega-6 and omega-9 fatty acids range from undetectable to detectable, and may even endure throughout subsequent cellular and organismic  generations  (Figure  3).  Although  research  affirms  a relationship between omega-3, omega-6 and omega-9 fatty acids, both synergistically with the metabolism of the other fatty acids, as well as individually in modulating specific pathways, findings are conflicting. Together the anti-inflammatory exertions,  along  with  the pro-inflammatory mechanisms, highlight the delicate, oftentimes calculated mercurial nature of these fatty acids in maintaining homeostasis.  Additional  research  is  needed  to  add  credence  to the

     

    emergence of omega-3, omega-6 and omega-9 fatty acids as modulators of metabolism, lipidomics and glycomics.

    Acknowledgement

    This work was supported by the Tuskegee University College Agriculture, Environment and Nutrition Sciences, the George Washington Carver Experiment Station and the Tuskegee University College of Arts and Sciences (Tuskegee, AL).

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