Neurochemical Research

, Volume 22, Issue 10, pp 1187–1192 | Cite as

Ethanol-Induced Cell Death by Lipid Peroxidation in PC12 Cells

  • Albert Y. Sun
  • Yong-mei Chen
  • Marilyn James-Kracke
  • Patricia Wixom
  • Yu Cheng


Free radical generation is hypothesized to be the cause of alcohol-induced tissue injury. Using fluorescent cis-parinaric acid and TBARS, lipid peroxidation was shown to be increased in the presence of trace amounts of free ferrous ion in PC12 cells. This increase in lipid peroxidation was enhanced by ethanol in a dose dependent manner and also correlated with loss of cell viability, as measured by increased release of lactate dehydrogenase (LDH). Resveratrol, a potent antioxidant, had a protective effect against lipid peroxidation and cell death. These findings strongly suggest that ethanol-induced tissue injury and cell death is a free radical mediated process, and may be important in alcohol-related premature aging and other degenerative diseases.

Lipid Peroxidation ethanol Cis-Parinaric acid = fluorescent probe for lipid peroxidation Resveratrol = antioxidant from grapes 


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  1. 1.
    DiLuzio, N. R. 1968 The role of lipid peroxidation and antioxidants in ethanol-induced lipid alteration. Exp. Mol. Pathol. 8:394–402.Google Scholar
  2. 2.
    Suematsu T., Matsumura T., Sato N., Miyamoto T., Ooka T., Kamada T., and Abe H. 1981. Lipid peroxidation in alcoholic liver disease in humans. Alcoholism 5:427–431.Google Scholar
  3. 2.
    Nordmann, R., Ribiere C., and Ravach H. 1992. Implication of free radical mechanisms in ethanol-induced cellular injury. Free Rad. Biol. Med. 12:219–240.Google Scholar
  4. 4.
    Montoliu, C., Valles, S., Renau-Piqueras, J., and Guerri, J. 1994. Ethanol induced oxygen radical formation and lipid peroxidation in rat brain: Effect of chronic alcohol consumption. J. Neurochem. 63:1855–1862.Google Scholar
  5. 5.
    Reinke, L. A., Lai, E. K., and McCay, P. B. 1988. Administration of ethanol or acetaldehyde to rats results in the generation of free radicals in liver, heart and other organs. Pages 663–666 in Kuriyama, K., Takada, A., and Ishii, H. (eds) Biochemical and Social Aspects of Alcohol and Alcoholism, Excerpta Medica, Amsterdam.Google Scholar
  6. 6.
    Ahmad, F. F., Cowan, K. L., and Sun, A. Y. 1988. Potentiation of ethanol induced lipid peroxidation of biological membranes by vitamin C. Life Sci. 43:1169–1176.Google Scholar
  7. 7.
    Albano, E., Tomase, A., Gona-Gatti, L., Poli, G., Vannini, V., and Kianzani, W. U. 1988. Free radical metabolism of ethanol. Free Rad. Res. Commun. 3:343–349.Google Scholar
  8. 8.
    Cheng, Y., Chen, Y., James-Kracke, M., Wixom, P., and Sun, A.Y. 1996. Enhanced lipid peroxidation by extracellular ATP in PC12 cells. Neurochem. Res. 21:27–33.Google Scholar
  9. 9.
    Renaud, S., and DeLorgeril, M. 1992. Wine alcohol, platelets and the French Paradox for coronary heart disease. Lancet 339: 1523–1526.Google Scholar
  10. 10.
    Frankel, E. N., Waterhouse, A. L., and Kinsella, J. E. 1993. Inhibition of human LDL oxidation by resveratrol. Lancet 341: 1103–1104.Google Scholar
  11. 11.
    Negre-Salvayre, A., and Salvayre, R. 1992 Quercetin prevents the cytotoxicity of oxidized LDL on lymphoid cell lines. Free Rad. Biol. Med. 12:101–106.Google Scholar
  12. 12.
    Morel, I., Lescoat, G., Cogrel, P., Sergent, O., Pasdeloup, N., Brissot, P., Cillard, P., and Cillard, J. 1993. Antioxidant and iron chelating activities of the flavonoids catechin, quercetin and diosmetin on iron loaded rat hepatocytes culture. Biochem. Pharmacol. 45: 13–19.Google Scholar
  13. 13.
    Fuhrman, B., Lavy, A., and Aviram, M. 1995. Consumption of red wine with meals reduces the susceptibility of human plasma and low density lipoprotein to lipid peroxidation. Am. J. Clin. Nutr. 61:549–554.Google Scholar
  14. 14.
    Jang, M., Cai, L., Udeani, G. O., Slowing, K. V., Thomas, C. F., Beecher C. W. W., Fong, H. H. S., Farnsworth N. R., Kinghorn, A. D., Mehta, R. G., Moon, R. C., and Pezzuto, J. M. 1997. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275:218–220.Google Scholar
  15. 15.
    Bergmeyer, H. U., and Brent, E. 1974. UV-assay with pyruvate and NADH. Pages 574–579 in Bergmeyer H. U. (ed) Methods of Enzymatic Analysis, Academic Press, Inc., New York.Google Scholar
  16. 16.
    Gutteridge, J. M. C. 1982. Free radical damage to lipids, amino acids, carbohydrates and nucleic acids determined by thiobarbituric acid reactivity. Int. J. Biochem. 14:649–653.Google Scholar
  17. 17.
    Cheng, Y., Wixom, P., James-Kracke, M. R., and Sun, A. Y. 1994. Effects of extracellular ATP on Fe++-induced cytotoxicity of PC12 cells. J. Neurochem. 63:895–902.Google Scholar
  18. 18.
    Sun, A. Y., Yang, W. I., and Kim, H. D. 1993. Free radical and lipid peroxidation in manganese-induced neuronal cell injury. Ann. New York Acad. Sci. 679:358–363.Google Scholar
  19. 19.
    May, P. M., Williams, D. R., and Linder, P. W. 1978. Biological significance of low molecular weight iron (III) complexes. Pages 29–76 in Sigel, H. (ed) Metal Ions in Biological Systems, Vol. 7. Marcel Dekker, Inc., New York.Google Scholar
  20. 20.
    Reinke, L. A., Lai, E. K., DuBose, C. M., and McCay, P. B. 1987. Reactive free radical generation in vivo in heart and liver of ethanol fed rats: correlation with radical formation in vitro. Proc. Nat. Acad. Sci. USA 84:9223–9227.Google Scholar
  21. 21.
    Knecht, K. T., Bradford, B. U., Mason R. P., and Thurman, R. G. 1990. In vivo formation of a free radical metabolite of ethanol. Mol. Pharmacol. 38:26–30.Google Scholar
  22. 22.
    Albano, E., Ciot I., Morimoto, M., Tomase, A., Angelman-Sundberg, M., and French, S. W. 1996. Role of cytochrome P450 2E1-dependent formation of hydroxyethyl free radical in the development of liver damage in rats intragastrically fed with ethanol. Hepatology 23:155–163.Google Scholar
  23. 23.
    Dianzani, M. U. 1985. Lipid peroxidation in ethanol poisoning: a critical reconsideration. Alcohol Alcoholism 20:161–173.Google Scholar
  24. 24.
    Shaw, S., Eng, J., and Jayatilleke, E. 1995. Ethanol-induced free radical injury to the hepatocyte glucagon receptor. Alcohol 12: 273–277.Google Scholar
  25. 25.
    Morgan, E. T., Koop, D. R., and Coon, M. J. 1982. Catalytic activity of cytochrome P-450 isozyme 3a isolated from liver microsomes of ethanol-treated rabbits: oxidation of alcohols. J. Biol. Chem. 257:13951–13957.Google Scholar
  26. 26.
    Winston, G. W., and Cederbaum, A. I. 1983. NADPH-dependent production of oxy radicals by purified components of the rat liver mixed function oxidase system. I. Oxidation of hydroxyl radical scavenging agents. J. Biol. Chem. 258:1508–1519.Google Scholar
  27. 27.
    Mira, L., Maia, L. Barreira, L., and Manso, C. F. 1995. Evidence for free radical generation due to NADH oxidation by aldehyde oxidase during ethanol metabolism. Arch. Biochem. Biophys. 318: 53–58.Google Scholar
  28. 28.
    Malmstrom, B. G. 1982. Enzymology of oxygen. Ann. Rev. Biochem. 51:21–59.Google Scholar
  29. 29.
    Shaw, S., and Jayatilleke, E. 1990. Ethanol-induced iron mobilization: role of acetaldehyde oxidase generated superoxide. Free Rad. Biol. Med. 9:11–17.Google Scholar
  30. 30.
    Perez, H. D., Roll, F. J., Bissell, D. M., Shak, S., and Goldstein, I. M. 1984. Production of chemotactic activity for polymorphonuclear leukocytes by cultured rat hepatocytes exposed to ethanol. J. Clin. Invest. 74:1350–1357.Google Scholar
  31. 31.
    Williams, A. J., and Barry, R. E. Free radical generation by neutrophils: a potential mechanism of cellular injury in acute alcoholic hepatitis. Gut 28:1157–1161.Google Scholar
  32. 32.
    Montoliu, C., Sancho-Tello, M., Azorin, I., Burgal, M., Vallis, S., Renau-Piqueras, J., and Guerri, C. 1995. Ethanol increases cytochrome P450 2E1 and induces oxidative stress in astrocytes. J. Neurochem. 65:2561–2570.Google Scholar
  33. 33.
    Halliwell, B., and Gutteridge, J. M. C. 1992. Biologically relevant metal ion-dependent hydroxyl radical generation—an update. FEBS Lett. 307:108–112.Google Scholar
  34. 34.
    Egan, T. J., Barthakur, S. R., and Aisen, P. 1992. Catalysis of the Haber-Weiss reaction by iron diethylenetriamine pentaacetate. J. Inorg. Biochem. 48:241–249.Google Scholar
  35. 35.
    Sun, A. Y., and Sun, G. Y. 1976. Functional roles of phospholipids of synaptosomal membranes. Adv. Exp. Med. Biol. 72:169–197.Google Scholar
  36. 36.
    Klatsky, A. L., Armstrong, M. A. and Friedman, G. D. 1992. Alcohol and Mortality. Ann. Intern. Med. 117:646–654.Google Scholar
  37. 37.
    Nanji, A. A., and French, S. W. 1986. Alcoholic beverages and coronary heart-disease. Atherosclerosis 60:197–198.Google Scholar
  38. 38.
    St. Leger, A. S., Cochrane, A. L., and Moore, F. 1979. Factors associated with cardiac mortality in developed countries with particular reference to the consumption of wine. Lancet 1:1017–1020.Google Scholar
  39. 39.
    Siemann, E. H., and Creasy, L. L. 1992. Concentration of the phytoalexin resveratrol in wine. Am. J. Enol. Vitic. 43:49–52.Google Scholar
  40. 40.
    Arichi, H., Kimura, Y., Okuda, H., Baba, K., Kozawa, K., and Arichi, S. 1982. Effects of stilbene components of the roots of polygonum Euspidatum Sieb et Zuce on lipid metabolism. Chem. Pharm. Bull. 30:1766–1779.Google Scholar
  41. 41.
    Kimura, Y., Ohminami, H., Okuda, H., Baba, K., Kozawa, M., and Arichi, S. 1983. Effects of stilbene components of roots of polygonum 550 on liver injury in peroxidized oil-fed rats. Plant Res. 49:51–54.Google Scholar
  42. 42.
    Kimura, Y., Okuda, H., and Arichi, S. Effects of stilbenes on arachidonate metabolism in leukocytes. Biochim. Biophys. Acta 834:275–278.Google Scholar
  43. 43.
    Chung, M. I., Teng, C. M., Cheng, K. L., Ko, F. N., and Lin, C. N. 1992. An anti-platelet principle of veratrum formosarum. Planta. Med. 58:274–276.Google Scholar
  44. 44.
    Noonberg, A., Goldstein, G., and Page, H. A. 1985. Premature aging in male alcoholics: “Accelerated aging” or “increased vulnerability”? Alcoholism Clin. Exp. Res. 9:334–338.Google Scholar

Copyright information

© Plenum Publishing Corporation 1997

Authors and Affiliations

  • Albert Y. Sun
    • 1
  • Yong-mei Chen
    • 1
  • Marilyn James-Kracke
    • 1
  • Patricia Wixom
    • 1
  • Yu Cheng
    • 2
  1. 1.Department of PharmacologyUniversity of MissouriColumbia
  2. 2.Department of NeurologyWashington UniversitySt. Louis

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