The effect of vitamin C and glutathione on ethanol cytotoxicity and selected parameters of pro- and antioxidative processes in mouse fibroblasts 3T3-L1

• Autorzy: Stanczyk M , Gromadzinska J. , Wasowicz W.
• Języki publikacji: EN

Abstrakty

EN

Ethanol metabolites may directly or indirectly induce oxidative stress as a result of disturbed balance between pro- and antioxidative processes. An indirect effect of ethanol on the generation of oxidative stress is associated with the attenuation of intracellular defensive enzymatic antioxidants, e.g. gluthatione peroxidase, catalase, superoxide dismutase as well as water- and/or fat-soluble low-molecular antioxidants (e.g., vitamin C, vitamin E, glutathione, selenium). The aim of our study was to evaluate the effect of 48h preincubation of mouse embryo fibroblast-like cells (3T3-L1) with vitamin C (VC, 0.06 mM) and glutathione (GSH, 0.05 mM) on their viability after exposure for 4, 8, and 24 h to ethanol (0.3 mM). Additionally, the activity of glutathione peroxidase (GSH- Px) and thioredoxin reductase (TrxR) enzymes as well as the level of thiobarbituric acid reactive substances (TBARS) were assessed in the exposed cells. In our study, vitamin C and glutathione had no significant effect on cytotoxicity of ethanol. We observed differences in GSH-Px and TrxR activity depending on the duration of exposure to ethanol. The highest GSH-Px activity in cell lysates was measured after 8 and 24h of incubation with ethanol and both antioxidants, or ethanol alone. The highest TrxR activity was observed after 4h of incubation in the presence of ethanol or GSH. There were no effects of VC and GSH on the activity of the antioxidant enzymes in comparison to cells not supplemented with VC and GSH. Similarly, we were unable to show any significant differences in levels of TBARS except incubation for 24h, when concentration of TBARS was highest in cells exposed to ethanol and simultaneously supplemented with the antioxidants.

Słowa kluczowe

EN

vitamin C; fibroblast-like cell; ethanol; mouse; mice; antioxidative process; glutathione; fibroblast; glutathione peroxidase; cytotoxicity; embryo

• Wydawca: -
• Czasopismo: Polish Journal of Environmental Studies
• Rocznik: 2006
• Tom: 15
• Numer: 1
• Opis fizyczny: p.131-137,fig.,ref.

Twórcy

autor

Stanczyk M

• Nofer Institute of Occupational Medicine, Teresy 9, 90-950 Lodz, Poland

autor

Gromadzinska J.

autor

Wasowicz W.

Bibliografia

• 1. SUSICK JR. L. R., ZANNONI V. G. Ascorbic acid and alcohol oxidation. Biochem. Pharmacol. 33, 3963, 1984.
• 2. DOPONT I., BODENEZ P., BERTHOU F., SIMON B., BARDOU L. G., LUCAS D. Cytochrome P-450 2E1 activity and oxidative stress in alcoholic patients. Alcohol Alcohol. 35, 98, 2000.
• 3. COHEN-KEREM R., KOREN G. Antioxidants and fetal protection against ethanol teratogenicity I. Review of the experimental data and implications to humans. Neurotoxicol. Teratol. 25, 1, 2003.
• 4. SIES H. Oxidative stress: from basic research to clinical application. Am. J. Med. 91, 31, 1991.
• 5. MANSOURI A., DEMEILLIERS CH., AMSELLEM S., PESSAYRE D., FROMENTY B. Acute ethanol administration oxidatively damages and depletes mitochondrial DNA in mouse liver, brain, heart, and skeletal muscles: protective effect of antioxidants. J. Pharmacol. Exp. Ther. 298, 737, 2001.
• 6. HOEK J. B., CAHILL A., PASTORINO J. G. Alcohol and mitochondria: a dysfunctional relationship. Gastroenterology 122, 2049, 2002.
• 7. ZHOU Z., SUN X., KANG Y. J. Metallothionein protection against alcoholic liver injury through inhibition of oxidative stress. Exp. Biol. Med. 227, 214, 2002.
• 8. MCDONOUGH K. H. Antioxidant nutrients and alcohol. Toxicology 189, 89, 2003.
• 9. BARTSCH H., NAIR J. Ultrasensitive and specific detection methods for exocyclic DNA adducts: Markers for lipid peroxidation and oxidative stress. Toxicology 153, 105, 2000.
• 10. NAVASUMRIT P., WARD T. H., DODD N. J. F., O’CONNOR P. J. Ethanol-induced free radicals and hepatic DNA strand breaks are prevented in vivo by antioxidants: effects of acute and chronic ethanol exposure. Carcinogenesis 21, 93, 2000.
• 11. JOUNG I. S., WOODSIDE J. V. Antioxidants in health and disease. J. Clin. Pathol. 54, 176, 2001.
• 12. DOBA T., BURTON G. W., INGLOT K. U. Antioxidant and co-antioxidant activity of vitamin C. The effect of vitamin C, either alone or in the presence of vitamin E or a water-soluble vitamin E analogue, upon the peroxidation of aqueous multilamellar phospholipid liposomes. Biochem. Biophys. Acta 835, 298, 1985.
• 13. CARR A., FREI B. Toward a new recommended dietary allowance for vitamin C based on antioxidant and health effects in humans. Am. J. Clin. Nutr. 69, 1086, 1999.
• 14. CHEPDA T., CADAU M., LASSABLIERE F., REYNAUD E., PERIER C., FREY J., CHAMSON A. Synergy between ascorbate and α-tocopherol on fibroblasts in culture. Life Sci. 69, 1587, 2001.
• 15. GINTER E., ZLOCH Z. Influence of vitamin C status on the metabolic rate of a single dose of ethanol-1⁻¹⁴ C in guinea pigs. Physiol. Res. 48, 369, 1999.
• 16. SUSICK JR R., ZANNONI V. G. Effect of ascorbic acid on the consequences of acute alcohol consumption in humans. Clin. Pharmacol. Ther. 41, 502, 1987.
• 17. HAMMOND CH. L., LEE T. K., BALLATORI N. Novel roles for glutathione in gene expression cell death and membrane transport of organics solvents. J. Hepatol. 34, 946, 2001.
• 18. ANNI H., PRISTATSKY P., ISRAEL Y. Binding of acetaldehyde to glutathione metabolite: mass spectrometric characterization of an acetaldehyde-cysteinylglycine conjugate. Alcohol. Clin. Exp. Res. 27, 1613, 2003.
• 19. KANBAGLI Ö., BALKAN J. AYKAC-TOKER G., UYSAL M. Hepatic mitochondrial prooxidant and antioxidant status in ethanol-induced liver injury in rats. Biol. Pharm. Bull. 25, 1482, 2002.
• 20. NORDMANN R. Alcohol and antioxidant systems. Alcohol Alcohol. 29, 513, 1994.
• 21. OZARAS R., TAHAN V., AYDIN S., UZUN H., KAYA S., SENTURK H. N-acetylcysteine attenuates alcohol-induced oxidative stress in the rat. World J. Gastroenterol. 9, 125, 2003.
• 22. HANSSEN M., NIELSEN S. E., BERG K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods 119, 203, 1989.
• 23. PAGLIA D.E., VALENTINE V.W. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J.Lab.Clin.Med. 70, 158-169, 1967.
• 24. SMITH A. D., MORRIS V. C., LEVANDER O. A. Rapid determination of glutathione peroxidase and thioredoxin reductase activities using a 96-well microplate format: comparison to standard cuvette based assays. Int.J.Vitam.Nutr. Res. 71, 87, 2001.
• 25. WĄSOWICZ W., NEVE J., PERETZ A. Optimized steps in fluorometric determination of thiobarbituric acid-reactive substances in serum: Importance of extraction pH and influence of sample preservation and storage. Clin.Chem. 39, 2522, 1993.
• 26. BRADFORD M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal.Biochem. 72, 248, 1976.
• 27. BJORNEBOE A., BJORNEBOE G. Antioxidant status and alcohol-related diseases. Alcohol Alcohol. 28, 111, 1993.
• 28. CELEC P., JÁNI P., SMREKOVÁ L., MRLIAN A., KÚDELA M., HODOSY J., BOOR P., KRISTOVÁ V., JAKUBOVSKÝ J., JEŽOVÁ D., HALČÁK J., BOŽEK P., SLÁMOVÁ J., ULIČNÁ O., HOJSIK D., JURKOVIČOVÁ I. Effects of anabolic steroids and antioxidant vitamins on ethanol-induced tissue injury. Life Sci. 74, 419, 2003.
• 29. HIRAISHI H., SHIMADA T., IVEY K., TERANO A. Role of antioxidant defenses against ethanol-induced damage in cultured rat gastric epithelial cells. J. Pharmacol. Exp. Ther. 289, 103, 1999.
• 30. GARRIGA J., ADANERO J., FERNÁNDEZ-SOLÁ J., URBANO-MÁRQUEZ A., CUSSÓ R. Ethanol inhibits skeletal muscle cell proliferation and delays its differentiation in cell culture. Alcohol Alcohol. 35, 236, 2000.
• 31. WOŹNIAK A., WOŹNIAK B., DREWA G., KASPRZAK H. A. The effect of ascorbic acid and ethanol on the level of thiobarbituric acid reactive substances (TBARS) in selected tissues of albino BALB/c mice. Biol. Pharm. Bull. 25, 943, 2002.
• 32. AHMAD F. F., COWAN D. L., SUN A. Y. Potentation of ethanol-induced lipid peroxidation of biological membranes by vitamin C. Life Sci 43, 1169, 1988.
• 33. SAKAGAMI H., SATOH K., HAKEDA Y., KUMEGAWA M. Apoptosis-inducing activity of vitamin C and vitamin K. Cell Mol Biol. 46, 129, 2000.
• 34. SAKAGAMI H., SATOH K. Modulating factors of radical intensity and cytotoxic activity of ascorbate. Anticancer Res. 17, 3513, 1997.
• 35. GINTER E., ZLOCH Z., ONDREIČKA R. Influence of vitamin C status on ethanol metabolism in guinea-pigs. Physiol. Res. 47, 137, 1998.
• 36. SÁNCHEZ-MORENO C., PANIAGUA M., MADRID A., MARTIN A. Protective effect of vitamin C against the ethanol mediated toxic effects on human brain glial cells. J. Nutr. Biochem. 14, 606, 2003.
• 37. LEVINE M., CONRY-CANTILENA C., WANG Y., WELCH R. W., WASHKO P. W., DHARIWAL K. R., PARK J. B., LAZAREV A., GRAUMLICH J. F., KING J., CANTILENA L. R. Vitamin C pharmacokinetics in healthy volunteers: evidence for a recommended dietary allowance. Proc. Natl. Acad. Sci. USA 93, 3704, 1996.
• 38. SMITH A. R., VISIOLI F., HAGEN T. M. Vitamin C matters: increased oxidative stress in cultured human aortic endothelial cells without supplemental ascorbic acid. FASEB Journal 16, 1102, 2002.
• 39. ARTEEL G. Oxidants and antioxidants in alcohol-induced liver disease. Gastroenterology 124, 778, 2003.