Canthaxanthin Research Articles

Suppression of mouse skin papilloma by canthaxanthin and beta-carotene in vivo: possibility of the regression of tumorigenesis by carotenoids without conversion to retinoic acid.

Using mouse skin papilloma as a model system, we examined whether the antitumorigenic activity of carotenoids was related to their provitamin A activity. Oral administration of canthaxanthin (CX) or beta-carotene at 200 mg/kg/day for 14 days significantly reduced the cumulative size of papillomas induced on the skin by 9,10-dimethyl-1,2-benzanthracene (p < 0.05), after the accumulation of these carotenoids in the tumors. The levels of a protooncogene, c-myc, were simultaneously suppressed in papillomas in carotenoid-treated mice. Because CX cannot be converted metabolically to retinoids, these results suggested that CX directly inhibited the growth of papillomas. Neither the accumulation of retinoids nor the expression of a retinoic acid-inducible gene, retinoic acid receptor-beta, was found in papillomas of CX- and beta-carotene-treated mice, suggesting that, like CX, beta-carotene might exert the tumor-suppressing effect without being converted to retinoids. Thus a certain antitumorigenic activity of carotenoids appears not necessarily to require their provitamin A activity.

Suppression of azoxymethane-induced rat colon carcinogenesis by dietary administration of naturally occurring xanthophylls astaxanthin and canthaxanthin during the postinitiation phase.

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.

Chemoprevention of mouse urinary bladder carcinogenesis by the naturally occurring carotenoid astaxanthin

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 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.

Serum carotenoids and retinoids in ferrets fed canthaxanthin

A high-performance liquid chromatographic method has been developed to analyze canthaxanthin (CX) with other carotenoids and retinoids in serum. Serum (100 μL) was extracted first with chloroform/methanol and then with hexane, using retinyl acetate and γ-carotene as the internal standards. The residue of the extract was resuspended into ethanol and injected onto an 8.3 × 0.46 cm 3-μm ODS column. The gradient system consisted of two solvents of acetonitrile/tetrahydrofuran/water with different ratios. The carotenoids and retinoids were measured at 450 nm and 340 nm, respectively. CX was detected in the serum of the ferrets fed CX (79% all- trans and 21% cis isomer, 50 mg/kg bodyweight by gavage for 1 month), but was not detected in the control group. The existence of CX in the serum was confirmed by its UV/visible absorption spectrum and by its negative chemical ionization/mass spectrum with an M − ion at m/z 564. The serum level of all- trans CX in the experimental ferrets was 33.34 ± 3.33 nmol/L ( n = 13) and that of cis CX was 72.48 ± 7.64 nmol/L ( n = 13). There was no difference between the CX-fed group and the control group in the serum levels of retinol, retinyl esters, lutein, cryptoxanthin, or β-carotene. Retinyl esters in ferret serum represent 93% of total vitamin A level, which is more than 10 times higher than the vitamin A level in human blood. High retinyl esters level in the circulation of the ferret is a general phenomenon in carnivores.

Dietary carotenoids influenced biochemical but not morphological changes in adult male rats fed a choline‐deficient diet

In a study of the effects of carotenoids, canthaxanthin (CA), beta-apo-8'-carotenal (BA), or beta-carotene in an extract of Spirulina-Dunaliella algae (AE) was fed at 0%, 0.1%, or 0.2% in a choline-deficient (CD) diet. In each of eight groups, 10 adult male Fischer 344 rats were fed diets with designated carotenoid sources and levels or a choline-sufficient diet for 12 weeks. Carotenoids altered some of the changes induced by the CD diet. Increases in enlargement of fatty livers and low plasma cholesterol levels occurred in rats fed 0.2% BA. Plasma retinol was further reduced 35% by BA or AE. BA and AE increased liver total vitamin A about 80% and 305%, respectively. Liver lipid peroxidation was enhanced and plasma alpha-tocopherol was reduced further by 1.0% AE. AE, BA, and CA (mg/g fat) depressed liver alpha-tocopherol about 49%, 67%, and 78%, respectively. The decreased liver alpha-tocopherol was concurrent with an increase in carotenoid stores of CA > BA > AE. Histopathological examination of sections of liver tissue by light microscopy showed fatty and cirrhotic changes in all rats fed CD diets. Histochemical evaluation based on a semiquantitative assay revealed a marked increase in peroxisome enzyme activity in the livers of all CD rats. None of the carotenoids appeared to have any effect on the development of morphological changes in the liver. Although carotenoids can function as antioxidants, they did not prevent changes observed in rats fed CD diets.

Free radicals as carcinogens and their quenchers as anticarcinogens

An oxygen dependent signal was detected, late in the 1950s by electron spin resonance (ESR) in a saline solution of hematoporphyrin (Hp) excited by light. This signal expressed a free radical consisting of 'some kind of an association between Hp and oxygen', that Smaller et al. called 'oxyradical' (HpOO.). It soon opened a new level of understanding in carcinogenesis triggered by photodynamic substances, including Hp itself, polycyclic hydrocarbons (PCHs), as well as any carcinogen involving molecular species activated by radiation and/or metabolic reaction. Early in the 1960s, this prompted the discovery of benzo(a)pyrene (BP) photocarcinogenic enhancement (BP-PCE) in mice, probably due to an increase in free oxygen radical generation following correct light exposure. This assumption was confirmed in 1980 by the fact that mice orally loaded with antioxidants and radical quenchers, such as beta-carotene (BC) and cantaxanthin (CX), were protected against BP-PCE at 100% and against total BP carcinogenicity at more than 60%. These achievements were presented as the bases of the current explosion of interest in biology and medicine in building up the new field of chemoprevention against cancer and other chronic diseases by supplementation with antioxidant vitamins, retinoids and especially carotenoids and their synergistic association. The relevant findings of this research obtained in the last decade in in vitro and in vivo experiments as well as human interventions are reported and discussed with personal contributions.

Effect of retinoids and carotenoids on prostaglandin formation by oral squamous carcinoma cells

Several studies have correlated the excessive production of prostaglandins (PGs) with tumor promotion and the suppression of the immune response. Inhibition of PGs by pharmacological agents has been demonstrated to enhance immunocompetence, and to suppress growth of tumors in animals and humans. In this study we examined the effect of retinol (I), all-trans-retinoic acid (II), N-(4-Hydroxyphenyl) retinamide (N-4-HPR) (III), canthaxanthin (CTX) (IV), and β-carotene (β-CT) (V) on the bioconversion of 14C-arachidonic acid (AA) to PGE 2 by squamous carcinoma cells of the tongue, SCC-25. Agents (I), (II), (III), (IV) inhibited while (V) stimulated PGE 2 formation in a dose related manner. N-4-HPR was the most potent inhibitor of PGE 2 synthesis. The data suggest that certain retinoids and carotenoids have the potential of inhibition of PG synthesis by oral squamous carcinoma cells. Inhibitory effects such as those described here and antioxidant properties might in part contribute to the antiinflammatory and anticarcinogenic activity of retinoids in vivo.

Effects of palm carotenoids in rat hepatic cytochrome P450-mediated benzo(a)pyrene metabolism

Using benzo(a)pyrene (BaP) metabolism as a probe for chemical carcinogenesis, in vitro and in vivo effects of palm-oil carotenoid [beta-carotene (BC), alpha-carotene (AC), or canthaxanthin (CTX)] on BaP metabolism in the rat hepatic cytochrome P450-mediated monooxygenase system were studied. Apparent Michaelis-Menten constants (Km) for formation of the precursor carcinogen, 7,8-dihydrodiol BaP, were found to be 14.4 (BC), 1.74 (AC), and 0.7 (CTX) mumol/L. The order of anticarcinogenic strength established in this study was BC much greater than AC greater than CTX. Increased formation of the detoxification intermediate, 3-hydroxy BaP, with increased carotenoid concentration was observed. The order of detoxification strength was BC greater than AC = CTX. The presence of carotenoids in vivo inhibited BaP metabolism. Using 9,10-dihydrodiol BaP as an indicator for inhibition, the order of the antioxidative activity was palm oil (with carotenoids) greater than BC greater than CTX greater than palm oil (without carotenoids). BC and AC may selectively modify the rat-liver microsomal enzymes, thus providing a biochemical mechanism for the inhibitory effect of palm carotenoids on chemical carcinogenesis.

Diverse carotenoids protect against chemically induced neoplastic transformation

The ability of diverse carotenoid to inhibit methylcholanthrene-induced transformation of 10T1/2 cells has been investigated. When delivered using tetrahydrofuran as a novel solvent, all carotenoids were absorbed by cultured cells. When continuously administered to methylcholanthrene-treated cultures 7 days after removal of the carcinogen, canthaxanthin, beta-carotene, alpha-carotene and lycopene inhibited the production of transformed foci in a dose-dependent manner in the above order of potency. This activity was not associated with drug toxicity or antiproliferative effects. Renierapurpurin and bixin did not inhibit transformation at concentrations less than or equal to 10(-5) M. Lutein was inhibitory at 10(-5) M, but was inactive at lower concentrations. Because of differences in stability in culture medium (alpha-carotene less than beta-carotene less than canthaxanthin less than lycopene less than lutein) and structure, cellular levels of drug differed up to 8-fold after administration of identical concentrations of compounds. Carotenoids with polar groups achieved highest cellular levels, however cellular uptake did not correlate with activity. For example, lutein, the most polar and most stable, reached the highest concentration in cells yet required a concentration of 10(-5) M for activity in the transformation assay, while alpha-carotene, the least stable and least concentrated by cells, was comparably active at 3 X 10(6) M. alpha-Tocopherol, a potent lipid-phase antioxidant, was as active as lycopene in the transformation assay but at a 10-fold higher concentration did not approach the activity of beta-carotene or canthaxanthin. Because the most potent of the carotenoids tested (i.e. beta-carotene, alpha-carotene, canthaxanthin) all have the potential for conversion to retinoids (though this has never been demonstrated in mammals for canthaxanthin), it is suggested that these compounds have two components to their action; one related to their antioxidant properties, the other to their pro-vitamin A activities.

Effects of the Consumption of Cigarettes, Alcohol and Foods on Serum Concentrations of Carotenoids, Retinol and Tocopherols in Healthy Inhabitants Living in a Rural Area of Hokkaido.

The relationship between serum concentrations of carotenoids, tocopherols and retinol and the consumption of cigarettes, alcohol and foods was investigated in 835 healthy inhabitants (aged 35-79 years) of a rural area of Hokkaido. Serum concentrations of beta-carotene (BC), alpha-carotene (AC), lycopene (LY), beta-cryptoxanthin (BX), zeaxanthin (including lutein, ZX), canthaxanthin (CX) and beta-tocopherol (including gamma-tocopherol, BT) were higher in females than in males, while serum retinol (RE) concentrations were lower. Serum alpha-tocopherol (AT) concentrations were the same for both sexes. The serum values of BC, BX, AC, LY and ZX for males were inversely associated with the consumption of cigarettes in multiple-regression analysis. The serum values of BC, AC and BX for males were inversely associated with alcohol consumption, while serum RE values were positively related to alcohol consumption for both sexes. There were significant relations between serum values of BC, AC, and ZX and the intake frequencies of carrots, tomatoes, milk and/or green-leaf vegetables, especially for females, and between serum BX values and the intake of oranges, juices and fruits. Serum RE and AT concentrations were not associated with any food intake, and BT concentrations were associated with the intake of tomatoes and juices for males.

Serum concentrations of carotenoids, retinol, and α-tocopherol in healthy persons determined by high-performance liquid chromatography

Serum concentrations of α-carotene (AC), β-carotene (BC), lycopene (LY), β-cryptoxanthin (CR), zeaxanthin (including lutein, ZX), canthaxanthin (CX), retinol (RE), and α-tocopherol (TO) in healthy persons (618 males and 1196 females) aged 7–86 years were determined by HPLC. Serum concentrations of BC among persons aged 20–49 years were higher with increasing age in females, but not in males. Serum CR concentrations decreased with age ranging from 7 to 39 years, while ZX concentrations rose in the age group of 20 to 59 years for both sexes. In contrast, serum RE concentrations and ratios of RE BC and RE CR , especially in males aged 20–49, were higher with age. Serum TO values in both sexes rose with age and were similar.

Cancer chemoprevention by supplemental carotenoids and synergism with retinol in mastodynia treatment

Cancer chemoprevention with beta-carotene (BC), canthaxanthin (CX) and retinol-BC is reported with respect to skin, breast, gastric, colon carcinogeneses induced by benzo(a)pyrene (BP) with or without ultra violet radiation (UV-A, UV-B), dimethylbenzathracene (DMBA) +/- UVB, P-UVA, N-methyl-N'-N-nitro-nitrosoguanidine (MNNG), dimethylhydrazine (DMH), and with respect to transplanted tumours. When animals were loaded with carotenoid supplementation one month before the carcinogenic induction (continued throughout the experiment), cancer prevention was observed up to 60-100%. The absence of provitamin A-activity in CX shows the carotenoid antioxidant property. Fifteen patients given BC + CX to prevent recurrences after radical removal of the primary neoplasia in organs like lung, breast, colon, urinary bladder, head and neck were studied in 1980-89. A longer than expected disease-free interval was preliminarily found. Supplementation of BC +/- retinol was also reported to prevent and treat oral leucoplakia. Supplementation and intermittent retinol administration was also tested in benign cyclical mastalgia with clear cut side effect free therapeutic results.

Enhancement of the Expression of Activation Markers on Human Peripheral Blood Mononuclear Cells by In Vitro Culture With Retinoids and Carotenoids

Retinoids (retinol, retinal, retinoic acid, retinyl beta-glucuronide, and 13-cis retinoic acid) and carotenoids (beta-carotene and canthaxanthin) were evaluated for their immunomodulatory effects on human peripheral blood T-lymphocyte subpopulations and natural killer (NK) cells. Peripheral blood mononuclear cells (PBMC) from healthy young volunteers were isolated and incubated for 72 hours at various levels of retinoids and carotenoids including a physiological concentration (10(-8) M). Expression of surface antigens for total T cells, T-helper and T-suppressor cells, and activation markers (transferrin receptor, HLA-Dr antigen, and interleukin 2 receptor) were analyzed with an EPICS V flow cytometer. Retinoic acid and 13-cis retinoic acid (13-cRA) produced significant increases in the percentage of cells with markers for total T-helper cells, with a minimal effect on percentage of lymphocytes with markers for NK cells. However, beta-carotene (BC), canthaxanthin (CTX), and retinyl beta-glucuronide (RBG) dramatically increased the percentage of PBMC with markers for NK cells and produced a smaller increase in lymphocytes with surface antigens identifying them as T-helper cells. Furthermore, retinol and retinal did not show significant change either in the percentage of lymphocytes with markers for T-helper cells or in the helper/suppressor ratio. An increase in the expression of HLA-Dr antigen and transferrin receptors was greater when cells were incubated with 13-cRA than with either BC, CTX, or RBG, while carotenoids produced a greater increase in the expression of IL-2 receptors than 13-cRA. Our study indicates that both retinoids and carotenoids might be activating different subpopulations of immune cells.

Antioxidant activity of dietary canthaxanthin

It has been suggested that canthaxanthin (CX), beta-carotene, and other carotenoids may inhibit carcinogenesis via antioxidant activity. CX has been shown to inhibit lipid peroxidation in liposomes; this experiment was designed to assess the antioxidant activity of CX in biological membranes. Chicks were fed semipurified diets supplemented with placebo beadlets or CX beadlets (5 g beadlets/kg diet; 0.5 g CX/kg diet) for five weeks. Diets were deficient in vitamin E and selenium to maximize the power of detecting an antioxidant effect of CX and to induce exudative diathesis, which is a vitamin E and selenium deficiency disease of chicks. Dietary CX had no effect on the onset, incidence, or severity of exudative diathesis. Liver homogenates from CX-fed chicks exhibited significantly (p = 0.02) decreased formation of thiobarbituric acid-reactive substances over time in ferrous ion-induced peroxidations. Because dietary CX increased hepatic alpha-tocopherol content, subsequent peroxidations were conducted in membrane fractions prepared from placebo and CX livers matched for alpha-tocopherol content. In this system, CX was marginally but inconsistently protective against peroxidation at both 15 torr O2 (p = 0.12) and 150 torr O2 (p = 0.07). Nanomolar changes in hepatic alpha-tocopherol, unlike CX, substantially altered rates of peroxidation. These results suggest that under these conditions, dietary CX increased resistance to lipid peroxidation primarily by enhancing membrane alpha-tocopherol levels and secondarily by providing weak direct antioxidant activity.

Cis-Canthaxanthin and other carotenoid-like compounds in canthaxanthin preparations

cis-Canthaxanthin and other carotenoid-like compounds in canthaxanthin preparations

Beta-carotene and canthaxanthin inhibit chemically- and physically-induced neoplastic transformation in 10T1/2 cells.

We have studied the effects of beta-carotene (beta-C), a vitamin A precursor of plant origin, and canthaxanthin (CTX), a non-provitamin A carotenoid, on the neoplastic transformation of C3H/10T1/2 murine fibroblast cells. Chemical transformation in this well-characterized cell system has previously been shown to be reversibly inhibited by retinoids, compounds with vitamin A-like activity. Here we show that both beta-C and CTX inhibit 3-methylcholanthrene (MCA)-induced transformation with ED50s of 9 x 10(-7) M and 2 x 10(-7) M, respectively. Both carotenoids failed to inhibit X-ray-induced transformation when the cells were treated prior to and during irradiation. However, when the drugs were added 1 week after X-irradiation and maintained in the medium thereafter, as in the chemical transformation protocol, both carotenoids inhibited subsequent development of transformed foci in a dose-dependent manner. Again, CTX was more effective than beta-C, such that 3 x 10(-6) M completely inhibited radiogenically-induced foci. Similar to the previously described action of retinoids, the inhibition of MCA-induced transformation was reversible; upon removal of the drug, transformed foci developed within 2 weeks, indicating that the carotenoids were not specifically toxic to initiated cells. Although both carotenoids caused a small dose-dependent decrease in the growth rate of both parental and initiated 10T1/2 cells, they did not markedly affect colony size or number when the cells were treated as in the transformation assays, nor did they influence the expression of neoplasia of two transformed cell lines. Although the actions of beta-C and CTX are similar to those of retinoids in the 10T1/2 system, we suggest that the carotenoids act via a different mechanism, since CTX cannot be converted to active retinoids in mammalian cells, and there is no evidence that 10T1/2 cells can convert beta-C to vitamin A. We suggest that the carotenoids' lipid anti-oxidant properties may be responsible for their inhibitory actions on transformation.

The Deposition of Synthetic Oxycarotenoids in Egg Yolks

SummaryThe relative efficiencies with which synthetic oxycarotenoids are deposited in egg yolks were examined in three experiments using either White Leghorn x Australorp (W x A) or White Leghorn (WL) hens. The oxycarotenoids measured were β-apo-8′-carotenoic acid ethyl ester (CAEE), canthaxanthin (CHX) and citranaxanthin (CTX). The effect of time and duration of administration of CAEE on its utilization was also measured.W x A hens, aged 36 weeks, deposited 30–46, 13–17 and 4.4-4.9% of the intake of CAEE, CHX and CTX, respectively in their egg yolks. The comparable values for W x A hens aged 53 weeks were 26–35, 17–18 and 3.2-3.3%, respectively.WL hens, aged 32 weeks, deposited 49–50 and 13–15% of the intake of CAEE and CHX, respectively.The deposition of CAEE tended to increase when it was given for 20 days rather than 10 days and when given daily at 15.00 h rather than 08.30 h.