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ASTAXANTHIN

44 ABSTRACTS

PubMed search of:

Astaxanthin, therapeutic = 16 hits

Astaxanthin, deficiency = 7 hits

Astaxanthin, human = 24 hits

Astaxanthin, animal = 143 hits

Astaxanthin, benefit = 1 hits

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Astaxanthin Overview

Astaxanthin is a fat-soluble, oxygenated pigment called a xanthophyll and a member of

the carotenoid family. It has a unique molecular structure that gives it powerful

antioxidant function. It is extracted from salmon, microalgae, and Pfaffia, a yeast.

Current research shows that due to astaxanthin’s potent antioxidant activity, it may be

beneficial in cardiovascular, immune, anti-inflammatory, and neurodegenerative diseases.

Specifically it inhibits lipid peroxidation at the cell level; crosses the blood-brain barrier,

effecting treatment of ocular, and neurodegenerative diseases such as glaucoma and

Alzheimer's; provides significantly more antioxidant capacity than other carotenoids and

antioxidants such as beta-carotene and Vitamin E; entraps free radicals by adding them to

its long, double-bonded chain rather than donating an electron; stabilizes the cell

membrane like a bridge because its polar end groups span the cell membrane, thus

increasing its rigidity and mechanical strength; neutralizes singlet and triplet oxygen (de-

charges) generated by UVA and UVB radiation and other sources; binds to a lipoprotein,

an efficient transport vehicle, making it more bioavailable; increases immune system

function including heightened production of antibody-secreting cells and Interleukin 2

and suppression of Interferon-gamma; inhibits reactive oxygen species that cause

inflammation; enhances the antioxidant actions of Vitamin E and Vitamin C and

encourages the release of Vitamin A from the liver when needed.

Astaxanthin has 100-500 times the antioxidant capacity of Vitamin E and 10 times the

antioxidant capacity of beta-carotene. Many laboratory studies also indicate astaxanthin

is a stronger antioxidant than lutein, lycopene and tocotrienols.

Dosage: The recommended dosage of 1 mg twice per day is similar on a weight basis to

current doses for beta-carotene and alpha-tocopherol.

Side Effects: There are no known side effects.

(Source: http://www.beta-glucan-info.com/astaxanthin-questions-answers.htm)

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Research Overview

1. Interferes with proinflammatory substances

2. Blocks nitric oxide enzyme activity

3. Is a powerful antioxidant

4. Astaxanthin bioavailability is increased by some long chain triglycerides

5. Protects beta-cell function in diabetes

6. Reduces glucose toxicity

7. Limits exercise-induced cardiac and skeletal damage in mice

8. Protects against DNA damage from UVA rays

9. Reduces stress-induced lipid peroxidation

10. Helps prevent atherosclerosis

11. Slows growth of H pylori infection

12. Inhibits tumor growth

13. Controls cancer cell proliferation in colon cancer

14. Controls cancer cell proliferation in bladder cancer

ASTAXANTHIN: 44 ABSTRACTS

1. Mol Cells. 2003 Aug 31;16(1):97-105.

Astaxanthin inhibits nitric oxide production and inflammatory gene expression by

suppressing I(kappa)B kinase-dependent NF-kappaB activation.

Lee SJ, Bai SK, Lee KS, Namkoong S, Na HJ, Ha KS, Han JA, Yim SV, Chang K, Kwon

YG, Lee SK, Kim YM.

Vascular System Research Center and Department of Molecular and Cellular

Biochemistry, Kangwon National University Biology, Chunchon 200-701, Korea.

Astaxanthin, a carotenoid without vitamin A activity, has shown anti-oxidant and

anti-inflammatory activities; however, its molecular action and mechanism have

not been elucidated. We examined in vitro and in vivo regulatory function of

astaxanthin on production of nitric oxide (NO) and prostaglandin E2 (PGE2) as

well as expression of inducible NO synthase (iNOS), cyclooxygenase-2, tumor

necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta). Astaxanthin

inhibited the expression or formation production of these proinflammatory

mediators and cytokines in both lipopolysaccharide (LPS)-stimulated RAW264.7

cells and primary macrophages. Astaxanthin also suppressed the serum levels of

NO, PGE2, TNF-alpha, and IL-1beta in LPS-administrated mice, and inhibited

NF-kappaB activation as well as iNOS promoter activity in RAW264.7 cells

stimulated with LPS. This compound directly inhibited the intracellular

accumulation of reactive oxygen species in LPS-stimulated RAW264.7 cells as well

as H2O2-induced NF-kappaB activation and iNOS expression. Moreover, astaxanthin

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blocked nuclear translocation of NF-kappaB p65 subunit and I(kappa)B(alpha)

degradation, which correlated with its inhibitory effect on I(kappa)B kinase

(IKK) activity. These results suggest that astaxanthin, probably due to its

antioxidant activity, inhibits the production of inflammatory mediators by

blocking NF-kappaB activation and as a consequent suppression of IKK activity

and I(kappa)B-alpha degradation.

2. Biochem Biophys Res Commun. 2003 Aug 1;307(3):704-12.

Direct superoxide anion scavenging by a disodium disuccinate astaxanthin

derivative: Relative efficacy of individual stereoisomers versus the statistical

mixture of stereoisomers by electron paramagnetic resonance imaging.

Cardounel AJ, Dumitrescu C, Zweier JL, Lockwood SF.

Davis Heart and Lung Research Institute, 473 West 12th Avenue, Columbus, OH

43210-1252, USA.

Carotenoids are a related group of greater than 600 natural compounds,

irrespective of geometric- and stereoisomers, with demonstrated antioxidant

efficacy. The carotenoids are broadly divided into "carotenes," or non-oxygen

substituted hydrocarbon carotenoids, and "xanthophylls," oxygen-substituted

carotenoids. The natural compounds are excellent singlet oxygen quenchers as

well as lipid peroxidation chain-breakers; this dual antioxidant capacity is

generally attributed to the activity of the polyene chain, and increases with

the number of conjugated double bonds along the polyene chain length. However,

the poor aqueous solubility of most carotenes and the vast majority of

xanthophylls limits their use as aqueous-phase singlet oxygen quenchers and

direct radical scavengers. A variety of introduction vehicles (e.g., organic

solvents, cyclodextrins) have been used to introduce the insoluble carotenoids

into aqueous test systems. Hawaii Biotech, Inc. (HBI) successfully synthesized a

novel carotenoid derivative, the disodium disuccinate derivative of astaxanthin

(3,3(')-dihydroxy-beta,beta-carotene-4,4(')-dione) in all-trans (all-E) form.

The novel derivative is a water-dispersible symmetric chiral molecule with two

chiral centers, yielding four stereoisomeric forms: 3R,3(')R and 3S,3(')S

(enantiomers), and the diastereomeric meso forms (3R,3(')S and 3(')R,3S). The

individual stereoisomers were synthesized at high purity (>90% by HPLC) and

compared directly for efficacy with the statistical mixture of stereoisomers

obtained from the synthesis from the commercial source of astaxanthin (1:2:1

ratio of 3S,3(')S, meso, and 3R,3(')R, respectively). Direct scavenging of

superoxide anion was evaluated in a standard in vitro isolated human neutrophil

assay by electron paramagnetic resonance (EPR) imaging, employing the spin-trap

DEPMPO. Each novel derivative was tested in pure aqueous formulation and in

ethanolic formulation shown to completely disaggregate the compounds in

solution. In each case, the ethanolic formulation was a more potent scavenging

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vehicle. No significant differences in scavenging efficiency were noted among

the individual stereoisomers and the statistical mixture of stereoisomers,

suggesting that the polyene chain alone was responsible for superoxide

scavenging. Dose-ranging revealed that the statistical mixture of stereoisomers

of the novel derivative, at millimolar (mM) concentrations, could nearly

completely eliminate the superoxide anion signal generated in the activated

human neutrophil assay. All ethanolic formulations of the novel derivatives

exhibited increased scavenging efficiency over equimolar concentrations of

non-esterified astaxanthin delivered in a dimethyl sulfoxide (DMSO) vehicle.

These novel compounds will likely find utility in applications requiring aqueous

delivery of a highly potent direct radical scavenger.

3. Eur J Pharm Sci. 2003 Jul;19(4):299-304.

Oral bioavailability of the antioxidant astaxanthin in humans is enhanced by

incorporation of lipid based formulations.

Mercke Odeberg J, Lignell A, Pettersson A, Hoglund P.

Department of Clinical Pharmacology, Lund University Hospital, S-221 85 Lund,

Sweden. johanna.odeberg@klinfarm.lu.se

Astaxanthin is a carotenoid with antioxidant properties, synthesised by plants

and algae, and distributed in marine seafood. Astaxanthin is also available as a

food supplement, but, like other carotenoids, is a very lipophilic compound and

has low oral bioavailability. However, bioavailability can be enhanced in the

presence of fat. There is not much information in the literature about the

pharmacokinetics of oral astaxanthin in humans. In this open parallel study,

healthy male volunteers received a single dose of 40 mg astaxanthin, as lipid

based formulations or as a commercially available food supplement, followed by

blood sampling for further analysis of plasma concentrations. Pharmacokinetic

parameters were calculated to evaluate the extent and rate of absorption from

each formulation. The elimination half-life was 15.9+/-5.3 h (n=32), and showed

a mono-phasic curve. Three lipid based formulations: long-chain triglyceride

(palm oil) and polysorbate 80 (formulation A), glycerol mono- and dioleate and

polysorbate 80 (formulation B), and glycerol mono- and dioleate, polysorbate 80

and sorbitan monooleate (formulation C), all showed enhanced bioavailability,

ranging from 1.7 to 3.7 times that of the reference formulation. The highest

bioavailability was observed with formulation B, containing a high content of

the hydrophilic synthetic surfactant polysorbate 80.

4. J Med Food. 2003 Spring;6(1):51-6.

Safety of an astaxanthin-rich Haematococcus pluvialis algal extract: a

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randomized clinical trial.

Spiller GA, Dewell A.

Health Research and Studies Center, Los Altos, CA 94023, USA. spiller@sphere.org

A growing body of scientific literature indicates that astaxanthin is a more

powerful antioxidant than other carotenoids and vitamin E and may confer

numerous health benefits. The purpose of this investigation was to conduct a

human safety study with a Haematococcus pluvialis algal extract with high levels

of astaxanthin. Thirty-five healthy adults age 35-69 years were enrolled in a

randomized, double-blind, placebo-controlled trial of 8 weeks' duration. All

participants took three gelcaps per day, one at each meal. Nineteen participants

received gelcaps with an algal extract in safflower oil, containing 2 mg of

astaxanthin each (treatment); 16 participants received gelcaps containing

safflower oil only (placebo). Blood pressure and blood chemistry tests,

including a comprehensive metabolic panel and cell blood count, were conducted

at the beginning of the trial and after 4 and 8 weeks of supplementation. No

significant differences were detected between the treatment and the placebo

groups after 8 weeks of supplementation with the algal extract in the parameters

analyzed, except for serum calcium, total protein, and eosinophils (P <.01).

Although the differences in these three parameters were statistically

significant, they were very small and are of no clinical importance. These

results reveal that 6 mg of astaxanthin per day from a H. pluvialis algal

extract can be safely consumed by healthy adults.

5. Invest Ophthalmol Vis Sci. 2003 Jun;44(6):2694-701.

Effects of astaxanthin on lipopolysaccharide-induced inflammation in vitro and

in vivo.

Ohgami K, Shiratori K, Kotake S, Nishida T, Mizuki N, Yazawa K, Ohno S.

Department of Ophthalmology and Visual Sciences, Hokkaido University Graduate

School of Medicine, Sapporo, Japan. kohgami@med.hokudai.ac.jp

PURPOSE: Astaxanthin (AST) is a carotenoid that is found in marine animals and

vegetables. Several previous studies have demonstrated that AST exhibits a wide

variety of biological activities including antioxidant, antitumor, and

anti-Helicobacter pylori effects. In this study, attention was focused on the

antioxidant effect of AST. The object of the present study was to investigate

the efficacy of AST in endotoxin-induced uveitis (EIU) in rats. In addition, the

effect of AST on endotoxin-induced nitric oxide (NO), prostaglandin E2 (PGE2),

and tumor necrosis factor (TNF)-alpha production in a mouse macrophage cell line

(RAW 264.7) was studied in vitro. METHODS: EIU was induced in male Lewis rats by

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a footpad injection of lipopolysaccharide (LPS). AST or prednisolone was

administered intravenously at 30 minutes before, at the same time as, or at 30

minutes after LPS treatment. The number of infiltrating cells and protein

concentration in the aqueous humor collected at 24 hours after LPS treatment was

determined. RAW 264.7 cells were pretreated with various concentrations of AST

for 24 hours and subsequently stimulated with 10 microg/mL of LPS for 24 hours.

The levels of PGE2, TNF-alpha, and NO production were determined in vivo and in

vitro. RESULTS: AST suppressed the development of EIU in a dose-dependent

fashion. The anti-inflammatory effect of 100 mg/kg AST was as strong as that of

10 mg/kg prednisolone. AST also decreased production of NO, activity of

inducible nitric oxide synthase (NOS), and production of PGE2 and TNF-alpha in

RAW264.7 cells in vitro in a dose-dependent manner. CONCLUSIONS: This study

suggests that AST has a dose-dependent ocular anti-inflammatory effect, by the

suppression of NO, PGE2, and TNF-alpha production, through directly blocking NOS

enzyme activity.

6. Trends Biotechnol. 2003 May;21(5):210-6.

Haematococcus astaxanthin: applications for human health and nutrition.

Guerin M, Huntley ME, Olaizola M.

Mera Pharmaceuticals Inc., 73-4460 Queen Kaahumanu Hwy, Suite 110, Kailua-Kona,

96740, Hawaii, USA

The carotenoid pigment astaxanthin has important applications in the

nutraceutical, cosmetics, food and feed industries. Haematococcus pluvialis is

the richest source of natural astaxanthin and is now cultivated at industrial

scale. Astaxanthin is a strong coloring agent and a potent antioxidant - its

strong antioxidant activity points to its potential to target several health

conditions. This article covers the antioxidant, UV-light protection,

anti-inflammatory and other properties of astaxanthin and its possible role in

many human health problems. The research reviewed supports the assumption that

protecting body tissues from oxidative damage with daily ingestion of natural

astaxanthin might be a practical and beneficial strategy in health management.

7. Redox Rep. 2002;7(5):290-3.

Astaxanthin protects beta-cells against glucose toxicity in diabetic db/db mice.

Uchiyama K, Naito Y, Hasegawa G, Nakamura N, Takahashi J, Yoshikawa T.

First Department of Medicine, Kyoto Prefectural University of Medicine, Kyoto,

Japan.

Page 7

Oxidative stress induced by hyperglycemia possibly causes the dysfunction of

pancreatic beta-cells and various forms of tissue damage in patients with

diabetes mellitus. Astaxanthin, a carotenoid of marine microalgae, is reported

as a strong anti-oxidant inhibiting lipid peroxidation and scavenging reactive

oxygen species. The aim of the present study was to examine whether astaxanthin

can elicit beneficial effects on the progressive destruction of pancreatic

beta-cells in db/db mice--a well-known obese model of type 2 diabetes. We used

diabetic C57BL/KsJ-db/db mice and db/m for the control. Astaxanthin treatment

was started at 6 weeks of age and its effects were evaluated at 10, 14, and 18

weeks of age by non-fasting blood glucose levels, intraperitoneal glucose

tolerance test including insulin secretion, and beta-cell histology. The

non-fasting blood glucose level in db/db mice was significantly higher than that

of db/m mice, and the higher level of blood glucose in db/db mice was

significantly decreased after treatment with astaxanthin. The ability of islet

cells to secrete insulin, as determined by the intraperitoneal glucose tolerance

test, was preserved in the astaxanthin-treated group. Histology of the pancreas

revealed no significant differences in the beta-cell mass between

astaxanthin-treated and -untreated db/db mice. In conclusion, these results

indicate that astaxanthin can exert beneficial effects in diabetes, with

preservation of beta-cell function. This finding suggests that anti-oxidants may

be potentially useful for reducing glucose toxicity.

8. J Pharm Sci. 2003 Apr;92(4):922-6.

Improved aqueous solubility of crystalline astaxanthin (3,3'-dihydroxy-beta,

beta-carotene-4,4'-dione) by Captisol (sulfobutyl ether beta-cyclodextrin).

Lockwood SF, O'Malley S, Mosher GL.

Hawaii Biotech, Inc., 99-193 Aiea Heights Drive, Suite 200, Aiea, Hawaii 96701,

USA. slockwood@hibiotech.com

Carotenoids are the most widely distributed natural pigments, with over 600

individual compounds identified and characterized from natural sources. A few

are commercially important molecules, having found utility as additions to

animal feed in the aquaculture, poultry, and swine feed industries. The majority

are lipophilic molecules with near zero inherent aqueous solubility. Many

different methods have been developed to make the carotenoids "water

dispersible," as true water solubility has not been described. Astaxanthin

(3,3'-dihydroxy-beta, beta-carotene-4,4'-dione) is a commercially important

oxygenated carotenoid that has gained wide acceptance as a feed additive in the

$50 billion salmon and trout aquaculture industry. Recently, interest in the

human health applications of astaxanthin has increased, with astaxanthin

receiving approval as a dietary supplement in several countries, including the

Page 8

United States. Moving astaxanthin into a pharmaceutical application will require

a chemical delivery system that overcomes the problems with parenteral

administration of a highly lipophilic, low molecular weight compound. In the

current study, the ability of sulfobutyl ether beta-cyclodextrin (sodium), as

the Captisol(R) brand, to increase the aqueous water solubility of crystalline

astaxanthin was evaluated. Complexation of crystalline astaxanthin with Captisol

increased the apparent water solubility of crystalline astaxanthin approximately

71-fold, to a concentration in the 2 microg/mL range. It is unlikely that this

increase in solubility will result in a pharmaceutically acceptable chemical

delivery system for humans. However, the increased aqueous solubility of

crystalline astaxanthin to the range achieved in the current study will likely

find utility in the introduction of crystalline astaxanthin into mammalian cell

culture systems that have previously been dependent upon liposomes, or toxic

organic solvents, for the introduction of carotenoids into aqueous solution.

Pharm Sci 92: 922-926, 2003

9. Antioxid Redox Signal. 2003 Feb;5(1):139-44.

Astaxanthin limits exercise-induced skeletal and cardiac muscle damage in mice.

Aoi W, Naito Y, Sakuma K, Kuchide M, Tokuda H, Maoka T, Toyokuni S, Oka S,

Yasuhara M, Yoshikawa T.

Department of Medicine, Kyoto Prefectural University of Medicine, Kyoto,

602-0841.

Dietary antioxidants may attenuate oxidative damage from strenuous exercise in

various tissues. Beneficial effects of the antioxidant astaxanthin have been

demonstrated in vitro, but not yet in vivo. We investigated the effect of

dietary supplementation with astaxanthin on oxidative damage induced by

strenuous exercise in mouse gastrocnemius and heart. C57BL/6 mice (7 weeks old)

were divided into groups: rested control, intense exercise, and exercise with

astaxanthin supplementation. After 3 weeks of exercise acclimation, both

exercise groups ran on a treadmill at 28 m/min until exhaustion.

Exercise-increased 4-hydroxy-2-nonenal-modified protein and

8-hydroxy-2'-deoxyguanosine in gastrocnemius and heart were blunted in the

astaxanthin group. Increases in plasma creatine kinase activity, and in

myeloperoxidase activity in gastrocnemius and heart, also were lessened by

astaxanthin. Astaxanthin showed accumulation in gastrocnemius and heart from the

3 week supplementation. Astaxanthin can attenuate exercise-induced damage in

mouse skeletal muscle and heart, including an associated neutrophil infiltration

Page 9

that induces further damage.

10. Comp Biochem Physiol C Toxicol Pharmacol. 2002 Nov;133(3):443-51.

Astaxanthin and canthaxanthin do not induce liver or kidney

xenobiotic-metabolizing enzymes in rainbow trout (Oncorhynchus mykiss Walbaum).

Page GI, Davies SJ.

Fish Nutrition Unit, Department of Biological Sciences, University of Plymouth,

Drake Circus, Plymouth PL4 8AA, UK. pagegi@mapleleaf.ca

This study was designed to assess the effects of dietary carotenoid

supplementation on liver and kidney xenobiotic-metabolizing enzymes in the

rainbow trout. Twelve rainbow trout (mean weight 266+/-10 g) were assigned to

each of three replicate tanks for each of four dietary treatments; astaxanthin,

canthaxanthin, negative control and positive control using beta-naphthoflavone,

at a target dietary inclusion of 100 mg kg(-1) for each additive. Fish were fed

for 3 weeks at a level of 1.2% body wt. day(-1). Serum carotenoid levels were

used as indicators of exposure and were not significantly different (P>0.05)

between carotenoid-fed trout. Livers and kidney were frozen separately in liquid

N(2) by immersion and microsomal fractions from pooled samples (n=3) assayed for

xenobiotic-metabolizing enzyme (cytochrome P450 monoxygenase) activities

including ethoxyresorufin O-deethylase; methoxyresorufin O-demethylase;

pentoxyresorufin O-dealkylase; benzoxyresorufin O-dearylase; and the conjugating

enzymes glucuronosyl transferase; and glutathione-s-transferase. Results

revealed that carotenoid treatment did not significantly (P>0.05) induce any

enzyme system examined. Results are discussed in the context of metabolism of

absorbed carotenoids.

11. J Dermatol Sci. 2002 Oct;30(1):73-84.

Modulatory effects of an algal extract containing astaxanthin on UVA-irradiated

cells in culture.

Lyons NM, O'Brien NM.

Department of Food Science, Food Technology and Nutrition, University College

Cork, Cork, Ireland. nob@ucc.ie

UV radiation from sunlight is the most potent environmental risk factor in skin

cancer pathogenesis. In the present study the ability of an algal extract to

protect against UVA-induced DNA alterations was examined in human skin

fibroblasts (1BR-3), human melanocytes (HEMAc) and human intestinal CaCo-2

Page 10

cells. The protective effects of the proprietary algal extract, which contained

a high level of the carotenoid astaxanthin, were compared with synthetic

astaxanthin. DNA damage was assessed using the single cell gel electrophoresis

or comet assay. In 1BR-3 cells, synthetic astaxanthin prevented UVA-induced DNA

damage at all concentrations (10 nM, 100 nM, 10 microM) tested. In addition, the

synthetic carotenoid also prevented DNA damage in both the HEMAc and CaCo-2

cells. The algal extract displayed protection against UVA-induced DNA damage

when the equivalent of 10 microM astaxanthin was added to all three-cell types,

however, at the lower concentrations (10 and 100 nM) no significant protection

was evident. There was a 4.6-fold increase in astaxanthin content of CaCo-2

cells exposed to the synthetic compound and a 2.5-fold increase in cells exposed

to algal extract. In 1BR-3 cells, exposure to UVA for 2 h resulted in a

significant induction of cellular superoxide dismutase (SOD) activity, coupled

with a marked decrease in cellular glutathione (GSH) content. However

pre-incubation (18 h) with 10 microM of the either the synthetic astaxanthin or

the algal extract prevented UVA-induced alterations in SOD activity and GSH

content. Similarly, in CaCo-2 cells a significant depletion of GSH was observed

following UVA-irradiation which was prevented by simultaneously incubating with

10 microM of either synthetic astaxanthin or the algal extract. SOD activity was

unchanged following UVA exposure in the intestinal cell line. This work suggests

a role for the algal extract as a potentially beneficial antioxidant.

12. Life Sci. 2002 Apr 21;70(21):2509-20.

Contribution of the antioxidative property of astaxanthin to its protective

effect on the promotion of cancer metastasis in mice treated with restraint

stress.

Kurihara H, Koda H, Asami S, Kiso Y, Tanaka T.

Institute for Health Care Science, Suntory Ltd., 1-1-1 Wakayamadai,

Shimamoto-cho, Mishima-gun, Osaka 618-8503, Japan.

Hiroshi_Kurihara@suntory.co.jp

We investigated the effects of astaxanthin on the antitumor effector activity of

natural killer (NK) cells suppressed by stress in mice in order to define the

immunological significance of astaxanthin (ASX) when combined with restraint

stress treatment. When the mice were treated with restraint stress alone, the

total number of spleen cells, and the level NK cell activity per spleen were

reduced to a nadir on day 3. The stress also caused a significant increase in

the lipid peroxidation of liver tissue. ASX (100 mg/kg/day, p.o., 4 days)

improved the immunological dysfunction induced by restraint stress. On the other

hand, metastatic nodules were observed in the livers of syngenic DBA/2 mice on

day 12 after inoculation of P815 mastocytoma cells. Hepatic metastasis was

promoted further by restraint stress when applied on day 3 before the

Page 11

inoculation of P815. Daily oral administration of ASX (1 mg/kg/day, p.o., 14

days) markedly attenuated the promotion of hepatic metastasis induced by

restraint stress. These results suggested that astaxanthin improves antitumor

immune responses by inhibiting of lipid peroxidation induced by stress.

13. Arch Toxicol. 2002 Jan;75(11-12):665-75.

Metabolism and CYP-inducer properties of astaxanthin in man and primary human

hepatocytes.

Kistler A, Liechti H, Pichard L, Wolz E, Oesterhelt G, Hayes A, Maurel P.

Vitamins and Fine Chemicals, Human Nutrition and Health, F. Hoffmann-La Roche

Ltd, Basel, Switzerland. kistlera@bluewin.ch

Previous investigations in the rat have shown that the non-provitamin A

carotenoid astaxanthin is metabolized into 3-hydroxy-4-oxo-beta-ionone and

3-hydroxy-4-oxo-7,8-dihydro-beta-ionone and, in addition, is a potent CYP1A gene

inducer. Here we investigated the metabolism of this compound as well as its

capacity to induce CYP genes in primary cultures of human hepatocytes. Free

metabolites of 14C-astaxanthin produced in this cellular model were purified by

high pressure liquid chromatography (HPLC) and identified by gas

chromatography-mass spectrometry (GC-MS) analyses as 3-hydroxy-4-oxo-beta-ionol

and 3-hydroxy-4-oxo-beta-ionone. In addition, deconjugation of polar compounds

by glusulase and further analyses with HPLC and GC-MS revealed four radiolabeled

metabolites including: 3-hydroxy-4-oxo-beta-ionol, 3-hydroxy-4-oxo-beta-ionone,

and their reduced forms, 3-hydroxy-4-oxo-7, 8-dihydro-beta-ionol and

3-hydroxy-4-oxo-7,8-dihydro-beta-ionone. The same four metabolites were

identified in human plasma from two volunteers who had orally taken 100 mg

astaxanthin 24 h before blood collection. In cultured hepatocytes, astaxanthin

was a significant inducer of the major cytochrome P450 enzyme, CYP3A4 as well as

of CYP2B6, but not of other CYPs, including those from CYP1A and CYP2C families.

The lack of autoinduction of astaxanthin metabolism in human hepatocytes

suggests that neither CYP3A4 nor CYP2B6 contribute to the formation of

metabolites. We conclude that metabolism of astaxanthin and its CYP-inducing

capacity are different in humans and in rats. The novel methodology used in our

studies could be extended to evaluating the role of metabolites of more

important carotenoids such as beta-carotene in differentiation and

carcinogenicity.

14. J Reprod Fertil Suppl. 2001;57:331-4.

Effect of supplementation with the antioxidant astaxanthin on reproduction,

pre-weaning growth performance of kits and daily milk intake in mink.

Page 12

Hansen KB, Tauson AH, Inborr J.

Department of Animal Science and Animal Health, Royal Veterinary and

Agricultural University, Gronnegardsvej 3, 1870 Frederiksberg C, Denmark.

The study comprised two parts. Firstly, the effects of dietary supplementation

with an algal meal (Novasta) with a high astaxanthin content on ovulation rate

(number of corpora lutea, implantation rate, number, mass and length of fetuses)

of breeding female mink were evaluated. Secondly, reproductive outcome (number

of live and stillborn kits), kit growth rate and milk intake were studied. Both

studies were performed on standard brown female mink (n = 20; control (n = 10)

and experimental (n = 10)) housed under conventional farm conditions.

Experimental animals were supplied with 5.35 mg astaxanthin per day (0.25 g

algal meal (Novasta)). The numbers of corpora lutea, implantation sites and

fetuses appeared to be higher in the group that was given astaxanthin but the

effect was not significant. The differences between treated and control mink

were 1.4 (corpora lutea), 0.9 (implantation sites) and 1.2 (litter size). The

percentage of stillborn kits was reduced by 6.3 (P < 0.005). The milk intake as

measured by use of the isotopic water dilution technique was not affected by

treatment group. Milk intake increased from about 19 g in week 1 of lactation to

about 30 g per kit per day in week 4 of lactation. Kit weight gain was not

affected by the experimental treatment.

15. Biochem Biophys Res Commun. 2001 Oct 19;288(1):225-32.

Astaxanthin and peridinin inhibit oxidative damage in Fe(2+)-loaded liposomes:

scavenging oxyradicals or changing membrane permeability?

Barros MP, Pinto E, Colepicolo P, Pedersen M.

Department of Botany, Stockholm University, SE-10691 Stockholm, Sweden.

mpbarros@botan.su.se

Astaxanthin and peridinin, two typical carotenoids of marine microalgae, and

lycopene were incorporated in phosphatidylcholine multilamellar liposomes and

tested as inhibitors of lipid oxidation. Contrarily to peridinin results,

astaxanthin strongly reduced lipid damage when the lipoperoxidation

promoters-H(2)O(2), tert-butyl hydroperoxide (t-ButOOH) or ascorbate-and

Fe(2+):EDTA were added simultaneously to the liposomes. In order to check if the

antioxidant activity of carotenoids was also related to their effect on membrane

permeability, the peroxidation processes were initiated by adding the promoters

to Fe(2+)-loaded liposomes (encapsulated in the inner aqueous solution). Despite

that the rigidifying effect of carotenoids in membranes was not directly

measured here, peridinin probably has decreased membrane permeability to

Page 13

initiators (t-ButOOH > ascorbate > H(2)O(2)) since its incorporation limited

oxidative damage on iron-liposomes. On the other hand, the antioxidant activity

of astaxanthin in iron-containing vesicles might be derived from its known

Press.

16. J Atheroscler Thromb. 2000;7(4):216-22.

Inhibition of low-density lipoprotein oxidation by astaxanthin.

Iwamoto T, Hosoda K, Hirano R, Kurata H, Matsumoto A, Miki W, Kamiyama M,

Itakura H, Yamamoto S, Kondo K.

National Institute of Health and Nutrition, Tokyo, Japan.

Marine animals produce astaxanthin which is a carotenoid and antioxidant. In

this study we determined the in vitro and ex vivo effects of astaxanthin on LDL

oxidation. The oxidation of LDL was measured in a 1 ml reaction system

consisting of increasing concentrations of astaxanthin (12.5, 25.0, 50.0

microg/ml), 400 microM V-70 (2, 2'-azobis(4-methoxy-2,

4-dimethylvaleronitrile)), and LDL (70 microg/ml protein). Astaxanthin dose,

dependently significantly prolonged the oxidation lag time (31.5, 45.4, 65.0

min) compared with the control (19.9 min). For the ex vivo study 24 volunteers

(mean age 28.2 [SD 7.8] years) consumed astaxanthin at doses of 1.8, 3.6,14.4

and 21.6 mg per day for 14 days. No other changes were made in the diet. Fasting

venous blood samples were taken at days 0, +14. LDL lag time was longer (5.0,

26.2, 42.3 and 30.7% respectively) compared with day 0 after consuming

astaxanthin at doses of 1.8, 3.6,14.4 and 21.6 mg for 14 days compared with day

0, but there was no difference in oxidation of LDL between day 0 (lag time

59.9+/-7.2 min) and day 14 (57.2+/-6.0 min) in the control group. Our results

provide evidence that consumption of marine animals producing astaxanthin

inhibits LDL oxidation and possibly therefore contributes to the prevention of

atherosclerosis.

17. Methods Find Exp Clin Pharmacol. 2001 Mar;23(2):79-84.

Effect of astaxanthin on the hepatotoxicity, lipid peroxidation and

Page 14

antioxidative enzymes in the liver of CCl4-treated rats.

Kang JO, Kim SJ, Kim H.

Department of Food and Nutrition, College of Human Ecology, Seoul National

University, Korea.

Astaxanthin is one of many carotenoids present in marine animals, vegetables and

fruits. Since carotenoids are known to have antioxidant properties, we tested to

determine if astaxanthin could have protective effects in the CCl4-treated rat

liver by activating the antioxidant system. Astaxanthin blocked the increase of

glutamate-oxalacetate transaminase (GOT) and glutamate-pyruvate transaminase

(GTP) activity and thiobarbituric acid reactive substances (TBARS) in response

to carbon tetrachloride (CCl4), while causing an increase in glutathione (GSH)

levels and superoxide dismutase (SOD) activities in the CCl4-treated rat liver.

These results suggest that astaxanthin protects liver damage induced by CCl4 by

inhibiting lipid peroxidation and stimulating the cellular antioxidant system.

18. Biochim Biophys Acta. 2001 Jun 6;1512(2):251-8.

Efficient radical trapping at the surface and inside the phospholipid membrane

is responsible for highly potent antiperoxidative activity of the carotenoid

astaxanthin.

Goto S, Kogure K, Abe K, Kimata Y, Kitahama K, Yamashita E, Terada H.

Faculty of Pharmaceutical Sciences, University of Tokushima, Japan.

Kogure@ph.tokushima-u.ac.jp

The effects of the carotenoids beta-carotene and astaxanthin on the peroxidation

of liposomes induced by ADP and Fe(2+) were examined. Both compounds inhibited

production of lipid peroxides, astaxanthin being about 2-fold more effective

than beta-carotene. The difference in the modes of destruction of the conjugated

polyene chain between beta-carotene and astaxanthin suggested that the

conjugated polyene moiety and terminal ring moieties of the more potent

astaxanthin trapped radicals in the membrane and both at the membrane surface

and in the membrane, respectively, whereas only the conjugated polyene chain of

beta-carotene was responsible for radical trapping near the membrane surface and

in the interior of the membrane. The efficient antioxidant activity of

astaxanthin is suggested to be due to the unique structure of the terminal ring

moiety.

Page 15

19. 0955-2863. 2000 Oct;11(10):482-490.

Plasma appearance and distribution of astaxanthin E/Z and R/S isomers in plasma

lipoproteins of men after single dose administration of astaxanthin(1).

Osterlie M, Bjerkeng B, Liaaen-Jensen S.

HIST, Department of Food Science, N-7004, Trondheim, Norway

Appearance, pharmacokinetics, and distribution of astaxanthin E/Z and R/S

isomers in plasma and lipoprotein fractions were studied in 3 middle-aged male

volunteers (37-43 years) after ingestion of a single meal containing a 100 mg

dose of astaxanthin. The astaxanthin source consisted of 74% all-E-, 9% 9Z-, 17%

13Z-astaxanthin (3R,3'R-, 3R,3'S; meso-, and 3S,3'S-astaxanthin in a 1:2:1

ratio). The plasma astaxanthin concentration--time curves were measured during

72 hr. Maximum levels of astaxanthin (1.3 +/- 0.1 mg/L) were reached 6.7 +/- 1.2

hr after administration, and the plasma astaxanthin elimination half-life was 21

+/- 11 hr. 13Z-Astaxanthin accumulated selectively, whereas the 3 and 3'R/S

astaxanthin distribution was similar to that of the experimental meal.

Astaxanthin was present mainly in very low-density lipoproteins containing

chylomicrons (VLDL/CM; 36-64% of total astaxanthin), whereas low-density

lipoprotein (LDL) and high-density lipoprotein (HDL) contained 29% and 24% of

total astaxanthin, respectively. The astaxanthin isomer distribution in plasma,

VLDL/CM, LDL, and HDL was not affected by time. The results indicate that a

selective process increases the relative proportion of astaxanthin Z-isomers

compared to the all-E-astaxanthin during blood uptake and that astaxanthin E/Z

isomers have similar pharmacokinetics.

20. Antimicrob Agents Chemother. 2000 Sep;44(9):2452-7.

Astaxanthin-rich algal meal and vitamin C inhibit Helicobacter pylori infection

in BALB/cA mice.

Wang X, Willen R, Wadstrom T.

Department of Infectious Diseases and Medical Microbiology, University of Lund,

Sweden.

Helicobacter pylori infection in humans is associated with chronic type B

gastritis, peptic ulcer disease, and gastric carcinoma. A high intake of

carotenoids and vitamin C has been proposed to prevent development of gastric

malignancies. The aim of this study was to explore if the microalga

Haematococcus pluvialis rich in the carotenoid astaxanthin and vitamin C can

inhibit experimental H. pylori infection in a BALB/cA mouse model. Six-week-old

BALB/cA mice were infected with the mouse-passaged H. pylori strain 119/95. At 2

Page 16

weeks postinoculation mice were treated orally once daily for 10 days (i) with

different doses of algal meal rich in astaxanthin (0.4, 2, and 4 g/kg of body

weight, with the astaxanthin content at 10, 50, and 100 mg/kg, respectively),

(ii) with a control meal (algal meal without astaxanthin, 4 g/kg), or (iii) with

vitamin C (400 mg/kg). Five mice from each group were sacrificed 1 day after the

cessation of treatment, and the other five animals were sacrificed 10 days after

the cessation of treatment. Culture of H. pylori and determination of the

inflammation score of the gastric mucosae were used to determine the outcome of

the treatment. Mice treated with astaxanthin-rich algal meal or vitamin C showed

significantly lower colonization levels and lower inflammation scores than those

of untreated or control-meal-treated animals at 1 day and 10 days after the

cessation of treatment. Lipid peroxidation was significantly decreased in mice

treated with the astaxanthin-rich algal meal and vitamin C compared with that of

animals not treated or treated with the control meal. Both astaxanthin-rich

algal meal and vitamin C showed an inhibitory effect on H. pylori growth in

vitro. In conclusion, antioxidants may be a new strategy for treating H. pylori

infection in humans.

21. J Nutr. 2000 Jul;130(7):1800-8.

Depletion of alpha-tocopherol and astaxanthin in Atlantic salmon (Salmo salar)

affects autoxidative defense and fatty acid metabolism.

Bell JG, McEvoy J, Tocher DR, Sargent JR.

Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, Scotland,

U.K.

Duplicate groups of Atlantic salmon post-smolts were fed four purified diets

supplemented with both vitamin E and the carotenoid astaxanthin (Ax) (+E, +Ax),

or supplemented with either vitamin E or Ax (-E, +Ax and +E, -Ax) or deficient

in both vitamin E and Ax (-E, -Ax) for 22 wk. There were no effects of diet on

growth rate, but an extensive lipoid liver degenerative lesion was observed in

15% of fish fed diets deficient in vitamin E. Tissue vitamin E concentrations

varied in accordance with dietary vitamin E in liver, muscle, heart, plasma,

brain and eye; levels were reduced to approximately 3% in liver but only to 40%

in eye of fish fed diets deficient in vitamin E compared with those fed diets

Page 17

supplemented with vitamin E. An interactive sparing of Ax supplementation on

tissue vitamin E concentration was observed, but only in brain. Dietary

deficiency of both vitamin E and Ax significantly increased the recovery of

desaturated and elongated products of both [1-(14)C] 18:3(n-3) and [1-(14)C]

20:5(n-3) in isolated hepatocytes, suggesting that conversion of fatty acids to

their long-chain highly unsaturated products can be stimulated by a deficiency

of lipid-soluble antioxidants. The antioxidant synergism of vitamin E and Ax was

supported by their ability to reduce malondialdehyde formation in an in vitro

stimulation of microsomal lipid peroxidation and to reduce plasma levels of

8-isoprostane. The results of this study suggest that both vitamin E and the

carotenoid Ax have antioxidant functions in Atlantic salmon.

22. Nutr Cancer. 2000;36(1):59-65.

Antitumor activity of astaxanthin and its mode of action.

Jyonouchi H, Sun S, Iijima K, Gross MD.

Department of Pediatrics, School of Medicine, University of Minnesota,

Minneapolis 55455, USA. jyono001@jyono001.email.umn.edu

Astaxanthin, a carotenoid without vitamin A activity, may exert antitumor

activity through the enhancement of immune responses. Here, we determined the

effects of dietary astaxanthin on tumor growth and tumor immunity against

transplantable methylcholanthrene-induced fibrosarcoma (Meth-A tumor) cells.

These tumor cells express a tumor antigen that induces T cell-mediated immune

responses in syngenic mice. BALB/c mice were fed astaxanthin (0.02%, 40

micrograms/kg body wt/day in a beadlet form) mixed in a chemically defined diet

starting zero, one, and three weeks before subcutaneous inoculation with tumor

cells (3 x 10(5) cells, 2 times the minimal tumorigenic dose). Three weeks after

inoculation, tumor size and weight were determined. We also determined cytotoxic

T lymphocyte (CTL) activity and interferon-gamma (IFN-gamma) production by

tumor-draining lymph node (TDLN) and spleen cells by restimulating cells with

Meth-A tumor cells in culture. The astaxanthin-fed mice had significantly lower

tumor size and weight than controls when supplementation was started one and

three weeks before tumor inoculation. This antitumor activity was paralleled

with higher CTL activity and IFN-gamma production by TDLN and spleen cells in

the astaxanthin-fed mice. CTL activity by TDLN cells was highest in mice fed

astaxanthin for three weeks before inoculation. When the

astaxanthin-supplemented diet was started at the same time as tumor inoculation,

none of these parameters were altered by dietary astaxanthin, except IFN-gamma

production by spleen cells. Total serum astaxanthin concentrations were

approximately 1.2 mumol/l when mice were fed astaxanthin (0.02%) for four weeks

and appeared to increase in correlation with the length of astaxanthin

supplementation. Our results indicate that dietary astaxanthin suppressed Meth-A

Page 18

tumor cell growth and stimulated immunity against Meth-A tumor antigen.

23. J Agric Food Chem. 2000 Apr;48(4):1150-4.

Antioxidant activities of astaxanthin and related carotenoids.

Naguib YM.

Phytochem Technologies, Chelmsford, MA 01824, USA.

The antioxidant activities of astaxanthin and related carotenoids have been

measured by employing a newly developed fluorometric assay. This assay is based

on 4,4-difluoro-3,5-bis(4-phenyl-1, 3-butadienyl)-4-bora-3a,4a-diaza-s-indacene

(BODIPY 665/676) as an indicator; 2,2'-azobis-2,4-dimethylvaleronitrile (AMVN)

as a peroxyl radical generator; and 6-hydroxy-2,5,7,

8-tetramethylchroman-2-carboxylic acid (Trolox) as a calibrator in an organic

and liposomal media. By employing this assay, three categories of carotenoids

were examined: namely, the hydrocarbon carotenoids lycopene, alpha-carotene, and

beta-carotene; the hydroxy carotenoid lutein; and the

alpha-hydroxy-ketocarotenoid astaxanthin. The relative peroxyl radical

scavenging activities of Trolox, astaxanthin, alpha-tocopherol, lycopene,

beta-carotene, lutein, and alpha-carotene in octane/butyronitrile (9:1, v/v)

were determined to be 1.0, 1.0, 1.3, 0.5, 0.4, 0.3, and 0.2, respectively. In

dioleoylphosphatidyl choline (DOPC) liposomal suspension in Tri-HCl buffer (pH

7.4 at 40 degrees C), the relative reactivities of astaxanthin, beta-carotene,

alpha-tocopherol, and lutein were found to be 1.00, 0.9, 0.6, and 0.6,

respectively. When BODIPY 665/676 was replaced by

4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,

4a-diaza-s-indacene-3-undecanoic acid (BODIPY 581/591 C(11)) as an indicator,

astaxanthin showed the highest antioxidant activity toward peroxyl radicals. The

relative reactivities of Trolox, astaxanthin, alpha-tocopherol, alpha-carotene,

lutein, beta-carotene, and lycopene were determined to be 1.0, 1.3, 0.9, 0.5,

0.4, 0.2, and 0.4, respectively.

24. Anticancer Res. 1999 Nov-Dec;19(6B):5223-7.

Dietary beta-carotene and astaxanthin but not canthaxanthin stimulate splenocyte

function in mice.

Chew BP, Wong MW, Park JS, Wong TS.

Department of Animal Sciences, Washington State University, Pullman 99164, USA.

The in vivo modulatory effect of beta-carotene, astaxanthin and canthaxanthin on

Page 19

lymphocyte function was investigated. Female BALB/c mice (8 wk old) were fed a

basal diet containing 0, 0.1% or 0.4% beta-carotene, astaxanthin or

canthaxanthin for 0, 2 or 4 wk (n = 8/diet/period). Splenic lymphocytes were

isolated and mitogen-stimulated proliferation, IL-2 production and lymphocyte

cytotoxicity were assessed. Body weight and feed intake were not different among

dietary treatments. Plasma carotenoids were undetectable in unsupplemented mice

but concentrations of the respective carotenoids were elevated in mice fed 0.1

or 0.4% beta-carotene (0.22 and 0.39 mumol/L), astaxanthin (16.4 and 50.2

mumol/L) and canthaxanthin (5.00 and 7.02 mumol/L) respectively. Mice fed both

dietary levels of beta-carotene and astaxanthin had enhanced

phytohemagglutinin-induced lymphoblastogenesis compared to unsupplemented mice

(P < 0.03). No treatment difference was detected with concanavalin A- or

lipopolysaccharide-induced lympho-proliferation nor with IL-2 production (P <

0.05). Astaxanthin (0.1%) also enhanced lymphocyte cytotoxic activity (P <

0.08). In contrast, canthaxanthin did not significantly influence any of the

lymphocyte functions measured. Results indicate that beta-carotene and

astaxanthin but not canthaxanthin exert enhanced splenic lymphocyte function in

mice.

25. Immunol Lett. 1999 Dec 1;70(3):185-9.

Treatment of H. pylori infected mice with antioxidant astaxanthin reduces

gastric inflammation, bacterial load and modulates cytokine release by

splenocytes.

Bennedsen M, Wang X, Willen R, Wadstrom T, Andersen LP.

Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark.

mbe@biobase.dk

Helicobacter pylori is a gram-negative bacterium affecting about half of the

world population, causing chronic gastritis type B dominated by activated

phagocytes. In some patients the disease evolves into gastric ulcer, duodenal

ulcer, gastric cancer or MALT lymphoma. The pathogenesis is in part caused by

the immunological response. In mouse models and in human disease, the mucosal

immune response is characterized by activated phagocytes. Mucosal T-lymphocytes

are producing IFN-gamma thus increasing mucosal inflammation and mucosal damage.

A low dietary intake of antioxidants such as carotenoids and vitamin C may be an

important factor for acquisition of H. pylori by humans. Dietary antioxidants

may also affect both acquisition of the infection and the bacterial load of H.

pylori infected mice. Antioxidants, including carotenoids, have

anti-inflammatory effects. The aim of the present study was to investigate

whether dietary antoxidant induced modulation of H. pylori in mice affected the

cytokines produced by H. pylori specific T-cells. We found that treatment of H.

pylori infected mice with an algal cell extract containing the antioxidant

Page 20

astaxanthin reduces bacterial load and gastric inflammation. These changes are

associated with a shift of the T-lymphocyte response from a predominant

Th1-response dominated by IFN-gamma to a Th1/Th2-response with IFN-gamma and

IL-4. To our knowledge, a switch from a Th1-response to a mixed Th1/Th2-response

during an ongoing infection has not been reported previously.

26. Anticancer Res. 1999 May-Jun;19(3A):1849-53.

A comparison of the anticancer activities of dietary beta-carotene,

canthaxanthin and astaxanthin in mice in vivo.

Chew BP, Park JS, Wong MW, Wong TS.

Department Animal Sciences, Washington State University, Pullman 99164-6320,

USA. boonchew@wsu.edu

The anticancer activities of beta-carotene, astaxanthin and canthaxanthin

against the growth of mammary tumors were studied in female eight-wk-old BALB/c

mice. The mice were fed a synthetic diet containing 0, 0.1 or 0.4%

beta-carotene, astaxanthin or canthaxanthin. After 3 weeks, all mice were

inoculated with 1 x 10(6) WAZ-2T tumor cells into the mammary fat pad. All

animals were killed on 45 d after inoculation with the tumor cells. No

carotenoids were detectable in the plasma or tumor tissues of unsupplemented

mice. Concentrations of plasma astaxanthin (20 to 28 mumol/L) were greater (P <

0.05) than that of beta-carotene (0.1 to 0.2 mumol/L) and canthaxanthin (3 to 6

mmol/L). However, in tumor tissues, the concentration of canthaxanthin (4.9 to

6.0 nmol/g) was higher than that of beta-carotene (0.2 to 0.5 nmol/g) and

astaxanthin (1.2 to 2.7 nmol/g). In general, all three carotenoids decreased

mammary tumor volume. Mammary tumor growth inhibition by astaxanthin was

dose-dependent and was higher than that of canthaxanthin and beta-carotene. Mice

fed 0.4% beta-carotene or canthaxanthin did not show further increases in tumor

growth inhibition compared to those fed 0.1% of each carotenoid. Lipid

peroxidation activity in tumors was lower (P < 0.05) in mice fed 0.4%

astaxanthin, but not in those fed beta-carotene and canthaxanthin. Therefore,

beta-carotene, canthaxanthin and especially astaxanthin inhibit the growth of

mammary tumors in mice; their anti-tumor activity is also influenced by the

supplemental dose.

27. Nutr Cancer. 1999;33(2):206-12.

Dietary lutein but not astaxanthin or beta-carotene increases pim-1 gene

expression in murine lymphocytes.

Park JS, Chew BP, Wong TS, Zhang JX, Magnuson NS.

Page 21

Department of Animal Sciences, Washington State University, Pullman 99164-6320,

USA.

This study investigates the effect of dietary carotenoids on pim-1 gene

expression in mouse splenocytes. Female BALB/c mice were fed 0%, 0.02%, or 0.4%

astaxanthin, beta-carotene, and lutein for two weeks. Plasma and liver were

obtained for the analysis of carotenoids. Splenocytes were isolated and cultured

in the presence of concanavalin A, and the level of pim-1 mRNA was determined by

Northern blot analysis. None of the carotenoids were detectable in the plasma

and liver of unsupplemented mice. In plasma the concentration of astaxanthin

(4.9-54.7 mumol/l) was dramatically higher than that of lutein (1.4-2.0 mumol/l)

and beta-carotene (0.1-0.7 mumol/l). Carotenoid uptake by the spleen but not the

liver reflected that observed in plasma. In mice fed 0.4% of each carotenoid,

the absolute concentration of the carotenoid in the liver was highest for

astaxanthin (24 nmol/g) followed by beta-carotene (7.5 nmol/g) and lutein (1.58

nmol/g). Mice fed lutein showed a dose-related increase in pim-1 mRNA

expression. The steady-state level of pim-1 mRNA in mice fed 0.4% lutein was

sixfold higher than in mice fed 0.02% lutein. In contrast, dietary astaxanthin

and beta-carotene did not affect pim-1 expression. Therefore, an increase in

pim-1 mRNA was observed in splenocytes stimulated with concanavalin A in

lutein-fed mice. This appears to be a unique effect of lutein and may be

associated with its antitumor activity observed in vivo.

28. Drug Metab Dispos. 1999 Apr;27(4):456-62.

Characterization of metabolites of astaxanthin in primary cultures of rat

hepatocytes.

Wolz E, Liechti H, Notter B, Oesterhelt G, Kistler A.

Vitamins and Fine Chemicals, Human Nutrition and Health, F. Hoffmann-LaRoche

Ltd., Basel, Switzerland. erich.wolz@roche.com

The metabolism of the nonprovitamin A carotenoid astaxanthin was investigated in

primary cultures of rat hepatocytes. In a time course study based on HPLC and

gas chromatography-mass spectrometry analyses, one main metabolite,

(rac)-3-hydroxy-4-oxo-beta-ionone, was found. This metabolite was conjugated

mainly into glucuronides, as demonstrated by glusulase treatment of the

conjugates under sulfatase-inhibiting conditions. Within 24 h more than 50%

astaxanthin was metabolized and conjugated. Deconjugation of the polar

conjugates with glusulase and analyses with HPLC and gas chromatography-mass

spectrometry identified two metabolites, (rac)-3-hydroxy-4-oxo-beta-ionone and

its reduced form (rac)-3-hydroxy-4-oxo-7,8-dihydro-beta-ionone, indicating that

the former was reduced in the conjugated form. We confirmed that the

Page 22

ketocarotenoid astaxanthin induces xenobiotic-metabolizing enzymes in rat liver

in vivo. However, there were no differences in the metabolism of astaxanthin in

cultured hepatocytes from rats that were pretreated with astaxanthin and, thus,

with induced cytochrome P-450 systems compared with control hepatocytes. Neither

liver microsomes from astaxanthin-pretreated nor control rats metabolized

astaxanthin. These results indicated that the cytochrome P-450 enzymes were not

involved in the metabolism of astaxanthin in rat hepatocytes. We conclude that

astaxanthin was metabolized in primary cultures of rat hepatocytes into

(rac)-3-hydroxy-4-oxo-beta-ionone and its reduced form

(rac)-3-hydroxy-4-oxo-7,8-dihydro-beta-ionone independent of the

xenobiotic-metabolizing enzymes induced by astaxanthin.

29. J Dermatol Sci. 1998 Mar;16(3):226-30.

Modulation of UVA light-induced oxidative stress by beta-carotene, lutein and

astaxanthin in cultured fibroblasts.

O'Connor I, O'Brien N.

Department of Nutrition, University College, Cork, Ireland.

The ability of beta-carotene, lutein or astaxanthin to protect against

UVA-induced oxidative stress in rat kidney fibroblasts (NRK) was assessed.

Activities of the antioxidant enzymes catalase (CAT) and superoxide dismutase

(SOD), and changes in thiobarbituric acid reactive substances (TBARS) were

measured as indices of oxidative stress. Exposure to UVA light at a dose

intensity of 5.6 mW/cm2 for 4 h resulted in a significant decrease in CAT and

SOD activities and a significant increase in TBARS. No cytotoxicity, as

indicated by lactate dehydrogenase (LDH) release, was observed. beta-Carotene (1

microM), lutein (1 microM) and astaxanthin (10 nM) protect against UVA

light-induced oxidative stress in vitro with astaxanthin exhibiting superior

protective properties.

30. Z Naturforsch [C]. 1998 Jan-Feb;53(1-2):93-100.

Does astaxanthin protect Haematococcus against light damage?

Fan L, Vonshak A, Zarka A, Boussiba S.

Microalgal Biotechnology Laboratory, Jacob Blaustein Institute for Desert

Research, Ben-Gurion University of the Negev, Israel.

The photoprotective function of the ketocarotenoid astaxanthin in Haematococcus

was questioned. When exposed to high irradiance and/or nutritional stress, green

Page 23

Haematococcus cells turned red due to accumulation of an immense quantity of the

red pigment astaxanthin. Our results demonstrate that: 1) The addition of

diphenylamine, an inhibitor of astaxanthin biosynthesis, causes cell death under

high light intensity; 2) Red cells are susceptible to high light stress to the

same extent or even higher then green ones upon exposure to a very high light

intensity (4000 mumol photon m(-2)s(-1)); 3) Addition of 1O2 generators

(methylene blue, rose bengal) under noninductive conditions (low light of 100

mumol photon m(-2)s(-1) induced astaxanthin accumulation. This can be reversed

by an exogenous 1O2 quencher (histidine); 4) Histidine can prevent the

accumulation of astaxanthin induced by phosphate starvation. We suggest that: 1)

Astaxanthin is the result of the photoprotection process rather than the

protective; 2) 1O2 is involved indirectly in astaxanthin accumulation process.

31. J Nutr Sci Vitaminol (Tokyo). 1997 Jun;43(3):345-55.

Inhibition of beta-carotene and astaxanthin of NADPH-dependent microsomal

phospholipid peroxidation.

Nakagawa K, Kang SD, Park DK, Handelman GJ, Miyazawa T.

Department of Applied Biological Chemistry, Tohoku University, Sendai, Japan.

To evaluate the antioxidant effects of beta-carotene and astaxanthin, rat liver

microsomes were exposed to a mixture of chelated iron (Fe3+/ADP) and NADPH. The

carotenoids (190 pmol/mg protein) were incorporated into some of these

microsomal membranes, and phospholipid hydroperoxides (PLOOH), thiobarbituric

acid reactive substances (TBARS) and endogenous alpha-tocopherol content were

measured over time after the initiation of oxidant stress. In control

microsomes, oxidant stress led to accumulation of 1,865 (+/- 371) pmol PLOOH/mg

protein during the initial 10-min peroxidation reaction, followed by a more

gradual decrease during the subsequent 20-min of reaction. PLOOH accumulation

during the initial 10-min reaction period was reduced to 588 (+/- 169) pmol/mg

protein with beta-carotene present and 800 (+/- 288) pmol/mg protein with

astaxanthin present. During the following 20-min of incubation, PLOOH levels

declined in the carotenoid-supplemented microsomes but continued to increase at

a slower rate in control preparations. TBARS did not show such large

accumulation as observed in PLOOH during the initial 10-min incubation in any

microsomal sample. The presence of carotenoids in the microsomal membrane

partially inhibited the loss of alpha-tocopherol, especially during the later

phase of oxidant stress. When lipid peroxidation is generated by membrane-bound

cyt-P450, the specific measurement of PLOOH clearly demonstrates that the

presence of carotenoids provides antioxidant protection.

32. Xenobiotica. 1996 Jan;26(1):49-63.

Page 24

Effects of canthaxanthin, astaxanthin, lycopene and lutein on liver

xenobiotic-metabolizing enzymes in the rat.

Gradelet S, Astorg P, Leclerc J, Chevalier J, Vernevaut MF, Siess MH.

Unite de Toxicologie Nutritionnelle, Institut National de la Recherche

Agronomique, DIJON, France.

1. The catalytic activities of several phase I and II xenobiotic-metabolizing

enzymes and the immunochemical detection of P4501A and 2B have been investigated

in liver microsomes and cytosol of male rats fed for 15 days with diets

containing canthaxanthin, astaxanthin, lycopene or lutein (as lutein esters)

(300 mg/kg diet) and in rats fed increasing levels (10, 30, 100 and 300 ppm) of

canthaxanthin or astaxanthin in the diet. 2. Canthaxanthin increased the liver

content of P450, the activities of NADH- and NADPH-cytochrome c reductase, and

produced a substantial increase of some P450-dependent activities, especially

ethoxyresorufin O-deethylase (EROD) (x 139) and methoxyresorufin O-demethylase

(MROD) (x 26). Canthaxanthin also increased pentoxy-(PROD) and benzoxyresorufin

O-dealkylases (BROD), but did not affect. NADPH-cytochrome c reductase and

erythromycin N-demethylase (ERDM) activities and decreased nitrosodimethylamine

N-demethylase (NDMAD) activity. Phase II p-nitrophenol UDP-glucuronosyl

transferase (4NP-UGT) and quinone reductase (QR) activities were also increased

by canthaxanthin treatment. These enhancing effects on EROD, MROD and 4NP-UGT

were clearly detectable at a dose as low as 10 ppm of canthaxanthin in the diet;

the induction of QR was only observed in rats fed > or = 100 ppm. Astaxanthin

induced the same pattern of enzymes activities as canthaxanthin, but to a lesser

extent: its effects on phase I enzymes and 4NP-UGT were observed in rats fed >

or = 100 ppm, and QR was not increased. Western blots of microsomal proteins

clearly showed the induction of P4501A1 and 1A2 by canthaxanthin and

astaxanthin. By contrast, lutein had no effect on the phase I and II

xenobiotic-metabolizing enzymes activities measured. Lycopene only decreased

NDMAD activity. 3. The two 4-oxocarotenoids canthaxanthin and astaxanthin are

substantial inducers of liver P4501A1 and 1A2 in the rat, and coinduce 4NP-UGT

and QR, just like polycyclic aromatic hydrocarbon, beta-naphtoflavone or dioxin

(TCDD). However, these latter classical P4501A inducers also induce aldehyde

dehydrogenase class 3 (ALDH3); this enzyme is not increased, or only marginally,

by canthaxanthin and astaxanthin. These two oxocarotenoids form a new class of

inducers of P4501A, are structurally very different from the classical inducers

quoted above, which are ligands of the AH receptor.

33. Carcinogenesis. 1995 Dec;16(12):2957-63.

Suppression of azoxymethane-induced rat colon carcinogenesis by dietary

administration of naturally occurring xanthophylls astaxanthin and canthaxanthin

Page 25

during the postinitiation phase.

Tanaka T, Kawamori T, Ohnishi M, Makita H, Mori H, Satoh K, Hara A.

First Department of Pathology, Gifu University School of Medicine, Japan.

The modulating effects of dietary feeding of two xanthophylls, astaxanthin (AX)

and canthaxanthin (CX) during the postinitiation phase on colon carcinogenesis

initiated with azoxymethane (AOM) were investigated in male F344 rats. Animals

were initiated with AOM by weekly s.c. injections of 15 mg/kg body wt for 3

weeks and then they were fed the diets containing AX or CX at concentrations of

100 and 500 p.p.m. for 34 weeks. The others contained the groups of rats treated

with AX or CX alone and untreated. At the end of the study (week 37), the

incidence and multiplicity of neoplasms (adenoma and adenocarcinoma) in the

large intestine of rats initiated with AOM and followed by AX or CX containing

diet at a high dose (500 p.p.m.) were significantly smaller than those of rats

given AOM alone (P < 0.001). In addition, AX or CX feeding significantly

inhibited the development of aberrant crypt foci induced by AOM. Dietary

exposure to AX or CX also decreased cell proliferation activity as revealed by

measuring 5'-bromodeoxyuridine-labeling index as crypt cells, colonic mucosal

ornithine decarboxylase activity and blood polyamine levels. These results

indicate that AX and CX are possible chemopreventers for carcinogenesis of colon

in addition to urinary bladder and oral cavity and such effects may be partly

due to suppression of cell proliferation.

34. J Nutr. 1995 Oct;125(10):2483-92.

Astaxanthin, a carotenoid without vitamin A activity, augments antibody

responses in cultures including T-helper cell clones and suboptimal doses of

antigen.

Jyonouchi H, Sun S, Tomita Y, Gross MD.

Department of Pediatrics, School of Medicine, University of Minnesota,

Minneapolis 55455, USA.

Astaxanthin, a carotenoid without vitamin A activity, enhances T-dependent

antigen (Ag)-specific humoral immune responses. We examined carotenoid actions

on T-helper (Th) cell activity in a direct manner with reconstitution

experiments; spleen Th cells were replaced with Ag-specific Type 1 and Type 2

(Th1 and Th2) Th cell clones. The Ag for the Th1 and Th2 clones were pigeon

cytochrome C and rabbit gamma-globulin, respectively. Astaxanthin and

beta-carotene augmented the number of IgM antibody (Ab)-secreting cells when

unprimed B cells were incubated with Th clones and stimulated with suboptimal

doses of Ag specific for each Th clone. The number of IgG Ab-secreting cells

Page 26

were greater with use of in vivo primed B cells than with unprimed B cells in

both Th clones. Astaxanthin but not beta-carotene augmented the number of IgG

Ab-secreting cells when primed B cells and Th cell clones were stimulated with

suboptimal doses of Ag specific for each Th clone. In the presence of optimal

doses of Ag for each Th clone, neither carotenoid augmented the number of

Ab-secreting cells. Astaxanthin and beta-carotene may enhance the actions of

both Th1 and Th2 cells for humoral immune responses with suboptimal Ag

challenges; certain carotenoids may help maintain Ag-mediated immune responses

at optimal levels.

35. Cancer Res. 1995 Sep 15;55(18):4059-64.

Chemoprevention of rat oral carcinogenesis by naturally occurring xanthophylls,

astaxanthin and canthaxanthin.

Tanaka T, Makita H, Ohnishi M, Mori H, Satoh K, Hara A.

First Department of Pathology, Gifu University School of Medicine, Japan.

The chemopreventive effects of two xanthophylls, astaxanthin (AX) and

canthaxanthin (CX) on oral carcinogenesis induced by 4-nitroquinoline 1-oxide

(4-NQO) was investigated in male F344 rats. Rats were given 20 ppm of 4-NQO in

their drinking water for 8 weeks to induce oral neoplasms or preneoplasms.

Animals were fed diets containing 100 ppm AX or CX during the initiation or

postinitiation phase of 4-NQO-induced oral carcinogenesis. The others contained

the groups of rats treated with AX or CX alone and untreated. At the end of the

study (week 32), the incidences of preneoplastic lesions and neoplasms in the

oral cavity of rats treated with 4-NQO and AX or CX were significantly smaller

than those of rats given 4-NQO alone (P < 0.001). In particular, no oral

neoplasms developed in rats fed AX and CX during the 4-NQO exposure and in those

given CX after the 4-NQO administration. Similarly, the incidences of oral

preneoplastic lesions (hyperplasia and dysplasia) in rats treated with 4-NQO and

AX or CX were significantly smaller than that of the 4-NQO-alone group (P <

0.05). In addition to such tumor inhibitory potential, dietary exposure of AX or

CX decreased cell proliferation activity in the nonlesional squamous epithelium

exposed to 4-NQO as revealed by measuring the silver-stained nucleolar organizer

regions protein number/nucleus and 5'-bromodeoxyuridine-labeling index. Also,

dietary AX and CX could reduce polyamine levels of oral mucosal tissues exposed

to 4-NQO. These results indicate that AX and CX are possible chemopreventers for

oral carcinogenesis, and such effects may be partly due to suppression of cell

proliferation.

36. Nutr Cancer. 1995;23(2):171-83.

Page 27

Effect of carotenoids on in vitro immunoglobulin production by human peripheral

blood mononuclear cells: astaxanthin, a carotenoid without vitamin A activity,

enhances in vitro immunoglobulin production in response to a T-dependent

stimulant and antigen.

Jyonouchi H, Sun S, Gross M.

Department of Pediatrics, School of Medicine, University of Minnesota,

Minneapolis 55455, USA.

The effect of carotenoids on in vitro immunoglobulin (Ig) production by

peripheral blood mononuclear cells (PBMNC) was examined by employing blood

samples from adult volunteers and full-term newborn babies (umbilical cord

blood). Under carotenoid-supplemented culture conditions, cells were stimulated

by polyclonal stimulants, neoantigens, and a recall antigen (Ag), and IgM, IgA,

and IgG levels in the culture supernatant were measured. Beta-carotene and

astaxanthin were used as representatives of carotenoids with and without vitamin

A activity, respectively. Astaxanthin enhanced IgM production in response to

T-dependent Ag (TD-Ag) and a T-dependent polyclonal stimulant. Astaxanthin also

augmented IgG production in response to a recall Ag. IgA production without

supplemental carotenoids was negligible for all stimuli. However, in

carotenoid-supplemented cultures, IgA production was significantly higher in

response to a T-dependent polyclonal stimulant than in unsupplemented cultures.

IgM and IgA production was augmented at 10(-8) mol/l astaxanthin, whereas

astaxanthin enhanced IgG production in response to a recall Ag at 10(-10)-10(-9)

mol/l. Similar enhancing actions of astaxanthin on IgM production were observed

in cord blood mononuclear cells (CBMNC), although CBMNC produced less IgM than

adult PBMNC. Beta-carotene did not have a significant effect on human Ig

production. The carotenoid actions were not demonstrated under serum-free

culture conditions; serum is essential for solubilization of carotenoids. In

summary, this study has shown for the first time that astaxanthin, a carotenoid

without vitamin A activity, enhances human Ig production in response to

T-dependent stimuli.

37. Int J Vitam Nutr Res. 1995;65(2):79-86.

Vitamin A status and metabolism of cutaneous polyamines in the hairless mouse

after UV irradiation: action of beta-carotene and astaxanthin.

Savoure N, Briand G, Amory-Touz MC, Combre A, Maudet M, Nicol M.

Biochimie Medicale A - Faculte de Medecine de Rennes, France.

Solar radiations (UV A and B) can cause epidermis photoaging and skin cancers.

These frequently irreversible effects result from the in situ generation of free

Page 28

radicals. However, it has been noted that nutritional factors can modulate

photochemical damage, in particular the common carotenoids present in food,

which can be considered as potential prophylactic agents against carcinogenesis.

We investigated the effect of UV A and B radiations on the skin of the SKH1

hairless mouse fed a diet either lacking in vitamin A or supplemented with

retinol, beta-carotene or astaxanthin. The latter is an oxygenated carotenoid

(like canthaxanthin) without provitamin A activity and with strong singlet

oxygen quenching ability. After analysing of vitamin status of each group

(plasma retinol concentrations and hepatic reserves), we searched for UV-induced

modifications of polyamine metabolism by measuring epidermal ornithine

decarboxylase (ODC) activity and free polyamines concentration (putrescine,

spermidine and spermine). In the basal state without irradiation, differences in

ODC activity between groups were nonsignificant; but after UV stimulation, ODC

increased markedly in the skin of vitamin A-deficient animals, much more than in

other groups. Curiously, the addition of astaxanthin or beta-carotene to the

regimen containing retinol reduced the protective effect of retinol alone.

Regarding polyamines after irradiation, putrescine was significantly increased

in the skin of deficient animals, in parallel with ODC activity. However,

astaxanthin had a stronger inhibitory effect on putrescine accumulation than

retinol, and decreased spermidine and spermine concentrations: this suggests a

specific action on transglutaminases.

38. Carcinogenesis. 1994 Jan;15(1):15-9.

Chemoprevention of mouse urinary bladder carcinogenesis by the naturally

occurring carotenoid astaxanthin.

Tanaka T, Morishita Y, Suzui M, Kojima T, Okumura A, Mori H.

First Department of Pathology, Gifu University School of Medicine, Japan.

The chemopreventive effects of two xanthophylls, astaxanthin (AX) and

canthaxanthin (CX), on urinary bladder carcinogenesis induced by

N-butyl-N(4-hydroxybutyl)nitrosamine (OH-BBN) was investigated in male ICR mice.

Mice were given 250 p.p.m. OH-BBN in drinking water for 20 weeks and after a 1

week interval with tap water, water containing AX or CX at a concentration of 50

p.p.m. was administered during subsequent 20 weeks. Other groups of mice were

treated with AX or CX alone or untreated. At the end of the study (week 41), the

incidences of preneoplastic lesions and neoplasms in the bladder of mice treated

with OH-BBN and AX or CX were smaller than those of mice given OH-BBN. In

particular, AX administration after OH-BBN exposure significantly reduced the

incidence of bladder cancer (transitional cell carcinoma) (P < 0.003). However,

the inhibition of the frequencies of such lesions in mice treated with OH-BBN

and CX was not significant. Treatment with AX or CX also decreased the

number/nucleus of silver-stained nucleolar organizer region proteins (AgNORs), a

Page 29

new index of cell proliferation, in the transitional epithelium exposed to

OH-BBN. Preneoplasms and neoplasms induced by OH-BBN, and the antiproliferative

potential, was greater for AX than CX. These results indicate that AX is a

possible chemopreventive agent for bladder carcinogenesis and such an effect of

AX may be partly due to suppression of cell proliferation.

39. J Nutr Sci Vitaminol (Tokyo). 1993 Dec;39(6):607-15.

Inhibitory effect of beta-carotene and astaxanthin on photosensitized oxidation

of phospholipid bilayers.

Oshima S, Ojima F, Sakamoto H, Ishiguro Y, Terao J.

Kagome Research Institute, Kagome Co., Ltd., Tochigi, Japan.

Large unilamellar liposomes comprising of egg yolk phosphatidylcholine (PC) was

exposed to photoirradiation in the presence of methylene blue (water-soluble

photosensitizer) or 12-(1-pyrene)dodecanoic acid (P-12, lipid-soluble

photosensitizer) to estimate the inhibitory effect of beta-carotene and

astaxanthin on photosensitized oxidation of phospholipid bilayers. Without

sensitizers, astaxanthin decreased much slower than beta-carotene and other

hydrocarbon carotenoids (lycopene, alpha-carotene). Astaxanthin lasted longer

than beta-carotene even in the presence of methylene blue or P-12. Decrease of

astaxanthin was also much slower than that of beta-carotene when egg yolk PC was

replaced by dimyristoyl PC. However, inhibitory effect of astaxanthin was lower

than beta-carotene in the case of P-12 sensitized photooxidation. These results

suggest that effectiveness of carotenoids as antioxidants on photosensitized

oxidation (Type II) in phospholipid bilayers depends on the site of singlet

oxygen to be generated, as well as their stability on photoirradiation.

40. Nutr Cancer. 1993;19(3):269-80.

Studies of immunomodulating actions of carotenoids. II. Astaxanthin enhances in

vitro antibody production to T-dependent antigens without facilitating

polyclonal B-cell activation.

Jyonouchi H, Zhang L, Tomita Y.

Department of Pediatrics, University of Minnesota, Minneapolis 55455.

Previously we have shown that astaxanthin, a carotenoid without provitamin A

activity, enhances in vitro antibody (Ab) production to sheep red blood cells in

normal B6 mice. In this study, we further attempted to examine the mechanisms of

this enhancing action of carotenoids on specific Ab production in vitro in

Page 30

relation to different antigen (Ag) stimuli, cytokine production, and T- and

B-cell interactions in both normal and autoimmune strains of mice. When the

actions of carotenoids were tested in normal strains of mice, we found that

astaxanthin enhanced in vitro Ab production to T cell-dependent Ag, but not to

T-independent Ag, and did not augment total immunoglobulin production.

Astaxanthin exerted maximum enhancing actions when it was present at the initial

period of Ag priming. This action of astaxanthin was abolished when T cells were

depleted from spleen cell suspensions and appeared to require direct

interactions between T and B cells. The results also indicated that carotenoids

may modulate the production of interferon-tau in this assay system. When the

actions of carotenoids were tested in autoimmune-prone MRL and NZB mice, the

enhancing action of astaxanthin on in vitro Ab production was less significant.

Furthermore, carotenoids did not potentiate or augment spontaneous Ab and

immunoglobulin production by spleen cells in these strains. Taken together,

carotenoids without provitamin A activity may be able to augment in vitro

specific Ab production to T cell-dependent Ag partly through affecting the

initial stage of Ag presentation without facilitating polyclonal B-cell

activation or autoantibody production.

41. Arch Latinoam Nutr. 1992 Dec;42(4):409-13.

[Hypercholesterolemic effect of canthaxanthin and astaxanthin in rats]

[Article in Spanish]

Murillo E.

Departamento de Quimica, Facultad de Ciencias Naturales y Exactas, Universidad

de Panama.

Three groups of male Wistar rats (130-140 g) were fed 30 days with a synthetic

diets containing 0.1% of beta-carotene, canthaxanthin and astaxanthin

respectively. Another group was fed with a synthetic diet without carotenoids.

The results shows that the beta-carotene does not induce change in plasma

cholesterol (49, 7 +/- 3.6 mg/dl), but canthaxanthin and astaxanthin induce a

significant increase in cholesterol concentration (92.1 +/- 3.6 and 66.5 +/- 5.1

mg/dl). This increase is noted mainly in the HDL fraction of the lipoproteins.

Canthaxanthin has more affinity than astaxanthin for the liver, principal site

of lipoproteins catabolism. The hypercholesterolemic effect of these

xanthophylls is not related to reported mechanisms of carotenoids in mammalian,

because beta-carotene does not induce changes in plasma cholesterol.

42. Arch Biochem Biophys. 1992 Sep;297(2):291-5.

Page 31

Astaxanthin and canthaxanthin are potent antioxidants in a membrane model.

Palozza P, Krinsky NI.

Department of Biochemistry, Tufts University School of Medicine, Boston,

Massachusetts 02111-1837.

When the conjugated keto-carotenoids, either astaxanthin or canthaxanthin, are

added to rat liver microsomes undergoing radical-initiated lipid peroxidation

under air, they are as effective as alpha-tocopherol in inhibiting this process.

This contrasts with the effect of beta-carotene, which is a much less potent

antioxidant when added in this system, without the addition of other

antioxidants.

43. Nutr Cancer. 1991;16(2):93-105.

Studies of immunomodulating actions of carotenoids. I. Effects of beta-carotene

and astaxanthin on murine lymphocyte functions and cell surface marker

expression in in vitro culture system.

Jyonouchi H, Hill RJ, Tomita Y, Good RA.

Department of Pediatrics, University of South Florida/All Children's Hospital,

St. Petersburg 33701.

The immunomodulating effects of carotenoids (beta-carotene and astaxanthin) on

mouse lymphocytes were studied in in vitro culture system by use of assay for

mitogen responses of spleen cells, thymocyte proliferation, interleukin 2

production, and antibody (Ab) production in vitro in response to sheep red blood

cells. Changes of cell surface markers on spleen lymphocytes including Ia

antigen (Ag), surface immunoglobulin, B220, and Thy-1 Ag were also examined. At

a concentration of 10(-8) M, carotenoids did not show any significant effect on

mitogen responses (phytohemagglutinin P and concanavalin A) on murine spleen

cells, irrespective of the concentrations of mitogens used. Interleukin 2

production by murine spleen cells was not significantly altered by carotenoids

in the culture media (10(-7) to 10(-9) M). [3H]thymidine incorporation by B6

thymocytes was somewhat enhanced in the presence of astaxanthin or beta-carotene

when cultured in the concentration of 10(6)/ml. At higher concentrations of

cells (5 x 10(6)/ml), such an effect was not observed. In assays of in vitro Ab

production in response to sheep red blood cells, B6 spleen cells produced

significantly more Ab-forming cells (plaque-forming cells, immunoglobulins M and

G) in the presence of astaxanthin (greater than 10(-8) M) but not beta-carotene.

Expression of Ia Ag seemed to be moderately enhanced on both Thy-1+ and Thy-1-

spleen cells in the presence of astaxanthin (greater than 10(-9) M) but not

beta-carotene. The expression of Thy-1 and surface immunoglobulin seemed

Page 32

unchanged with the treatment of these carotenoids. These results indicate that

immunomodulating actions of carotenoids are not necessarily related to

provitamin A activity, because astaxanthin, which does not have provitamin A

activity, showed more significant effects in these bioassays and also indicate

that such actions of carotenoid demonstrated in this study may be difficult to

explain only by its oxygen-quenching capacity.

44. Physiol Chem Phys Med NMR. 1990;22(1):27-38.

Inhibition of oxidative injury of biological membranes by astaxanthin.

Kurashige M, Okimasu E, Inoue M, Utsumi K.

Department of Medical Biology, Kochi Medical School, Japan.

The value of astaxanthin, a carotenoid pigment, in the treatment of oxidative

injury is assessed. Astaxanthin protects the mitochondria of vitamin E-deficient

rats from damage by Fe2(+)-catalyzed lipid peroxidation both in vivo and in

vitro. The inhibitory effect of astaxanthin on mitochondrial lipid peroxidation

is stronger than that of alpha-tocopherol. Thin layer chromatographic analysis

shows that the change in phospholipid components of erythrocytes from vitamin

E-deficient rats induced by Fe2+ and Fe3(+)-xanthine/xanthine oxidase system was

significantly suppressed by astaxanthin. Carrageenan-induced inflammation of the

paw is also significantly inhibited by administration of astaxanthin. These data

indicate that astaxanthin functions as a potent antioxidant both in vivo and in

vitro.