Effect of Herbal Extracts Mixtures on Antioxidant System in Chronic Enthanol-treated Rats

  • Kim, Mok-Kyung (Department of Hygienic Chemistry and Kyung Hee East-West Pharmaceutical Research Institute, College of Pharmacy, Kyung Hee University) ;
  • Won, Eun-Kyung (Department of Hygienic Chemistry and Kyung Hee East-West Pharmaceutical Research Institute, College of Pharmacy, Kyung Hee University) ;
  • Choung, Se-Young (Department of Hygienic Chemistry and Kyung Hee East-West Pharmaceutical Research Institute, College of Pharmacy, Kyung Hee University)
  • Published : 2006.12.30


Disturbance of antioxidant system is very common in chronic alcoholics and herbal or natural products with antioxidant activity have been used for its treatment. This study was to investigate the effect of Vitis vinifera extract(V), Schisandra chinensis extract(S), Taraxacum officinale extract(T), Gardenia jasminoides extract(G), Angelica acutiloba extract(A) and Paeonia japonica extract(P), and their combinations on the antioxidant and ethanol oxidation system. Male Sprague-Dawley rats were subjected to Lieber-DeCarli ethanol liquid diet(ED) and were then given different herbal extract mixtures for 6 weeks including VST(V 100+S 150+T 150mg/kg/day), VSG(V 100+S 150+G 150mg/kg/day), VTG(V 100+T 150+G 150mg/kg/day), and VAP(V 100+A 150+P 150mg/kg/day). When the activity of alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase(ALDH) were compared between ED only group and herbal extracts treatment group, the differences were statistically significant. Phase I and II(glutathione-S-transferase, phenol sulfatransferase) enzyme activities were found to be significantly higher in the VAT treatment group compared to the ED group. Herbal extracts not only repressed the ethanol-induced elevation of malondialdehyde level, but also protected against ethanol-induced decrease in glutathione content, glutathione reductase, glutathione peroxidase, catalase and superoxide dismutase activities. The administration of the herbal extracts was found to be effective in eliminating lipid-peroxides induced by long-term consumption of alcohol by activating various enzyme systems and physiological active compound formation system. After a chronic consumption of alcohol, Angelica Radix protected the liver via activating the ethanol-metabolism enzyme system, and Paeoniae Radix via activating the ethanol-metabolism enzyme and the phase I, II-metabolism enzyme system. Taraxaci Herba was also effective in liver protection via activating the ethanol-metabolism enzyme system and the phase I, II-metabolism enzyme system, Gardeniae Fructus via activating the phase II-metabolism enzyme system and the anti-oxidation system enzyme, and Schisandra Fructus and a grapestone via activating the anti-oxidation system. Our data suggest that these herbal extracts may be useful as a health functional food or new drug candidate for fatty liver and hepatotoxicity induced by chronic alcohol consumption.



  1. Aebi H., (1984). Catalase in vitro. Methods EnzymoI. 105, 121-126 https://doi.org/10.1016/S0076-6879(84)05016-3
  2. Albano E., (2006). Alcohol, oxidative stress and free radical damage. Proc Nutr Soc. 65, 278-290
  3. Aykac G., Dysal M., and Yalcin A. S., (1985). The effect of chronic ethanol ingestion on hepatic lipid peroxide, glutathione, glutathione peroxidase and glutathione transferase in rats. Toxicology 36, 71-76 https://doi.org/10.1016/0300-483X(85)90008-3
  4. Bergmyer H. U., (1974). Methods of enzymeatic analysis. Academic press, New York, 28
  5. Bradford M. M., (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72, 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  6. Caballeria J., Frezza M., and Hernandez-Munoz R. (1989). Gastric origin of the first-pass metabolism of ethanol in humans: effect of gastrectomy. Gastroenterology 97, 1205-1209 https://doi.org/10.1016/0016-5085(89)91691-0
  7. Carlberg I., and Mannervik B., (1975). Purification and characterization of the flavoenzyme glutathione reductase from rat liver. J Biol Chem. 250, 5475-5480
  8. Cederbaum A. I., (1989). Role of lipid peroxidation and oxidative stress in alcohol toxicity. Free Radic Biol Med. 7, 537-539 https://doi.org/10.1016/0891-5849(89)90029-4
  9. Cho M. H., Shim S. M., and Lee S. R., (2005). Effect of Evodiae fructus extracts on gene expressions related with alcohol metabolism and antioxidation in ethanol-loaded mice. Food Chem Toxicol. 43, 1365-1371 https://doi.org/10.1016/j.fct.2005.03.010
  10. Comporti M., (1993).Lipid peroxidation : An overview. In : Poli, G., albino, E., Dianzani, M.U., ets. Free Radicals: From Basic Science to Medicine(Molecular and cell Biology updates). Birkhauser Verag, Basel, Switzerland. 65
  11. Habig W. H., Pabst M. J., and Jakoby W. B.. (1974). Glutathione Svtransferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 249, 7130-7139
  12. Howard J.A., (1972). Absolute rate contents for reaction of oxygen radicals. Adv. Free Radical Chem. 4, 49
  13. ISSELBACHER K. J., CHRABAS M. F., and QUINN R. C., (1962). The solubilization and partial purification of a glucuronyl transferase from rabbit liver microsomes. J Biol Chem. 237, 3033-3036
  14. Julkunen R. J., Di Padova C., and Lieber C. S., (1985). First pass metabolism of ethanol--a gastrointestinal barrier against the systemic toxicity of ethanol. Life Sci. 37, 567-573 https://doi.org/10.1016/0024-3205(85)90470-9
  15. Jung S.Y., (2000). Effects of Leucocyanidin on aceta. minopheninduced hepatotoxicity and the mechanism. Masteres thesis
  16. Khan S. G., Katiyar S. K., and Agarwal R., (1992). Enhancement of antioxidant and phase II enzymes by oral feeding of green tea polyphenols in drinking water to SKH-1 hairless mice: possible role in cancer chemoprevention. Cancer Res. 52, 4050-4052
  17. Kim M. K., Lee Y. H., and Choung S. Y., (2006). Effects of herbal extracts on serum and liver lipid levels in chronic ethanol administered rats. Yakhak Hoeji. 49, 477-483
  18. Kim M. K., Hyun S. H., and Choung S. Y., (2006). Effects of Chronic ethanol consumption and herbal extracts administration on the antioxidant system and ethanol oxidation system in rats. Yakhak Hoeji. 50, 4. 245-262
  19. Klouckova I., Hrncirova P., and Mechref Y. (2006). Changes in liver protein abundance in inbred alcohol-preferring rats due to chronic alcohol exposure, as measured through a proteomics approach. Proteomics. 6, 3060-3074 https://doi.org/10.1002/pmic.200500725
  20. Koivula T., and Lindros K. O., (1975). Effects oflong-term ethanol treatment on aldehyde and alcohol dehydrogenase activities in rat liver. Biochem Pharmacol, 24, 1937-1942 https://doi.org/10.1016/0006-2952(75)90378-0
  21. Koop D. R., (1986). Hydroxylation of p-nitrophenol by rabbit ethanol-inducible cytochrome P-450 isozyme 3a. Mol Pharmacol. 29, 399-404
  22. Lieber C. S., (1994). Alcohol and the liver: 1994 update. Gastro-enterology 106, 1085-1105 https://doi.org/10.1016/0016-5085(94)90772-2
  23. Lieber C. S., and DeCarli L. M., (1968). Ethanol oxidation by hepatic microsomes: adaptive increase after ethanol feeding. Science 162, 917-918 https://doi.org/10.1126/science.162.3856.917
  24. Lieber C. S., and DeCarli L. M., (1982). The feeding of alcohol in liquid diets: two decades of applications and 1982 update. Alcohol Clin Exp Res. 6, 523-531 https://doi.org/10.1111/j.1530-0277.1982.tb05017.x
  25. Lin K. J, Chen J. C, and Tsauer W. (2001). Prophylactic effect of four prescriptions of traditional Chinese medicine on alphanaphthylisothiocyanate and carbon tetrachloride induced toxicity in rats. Acta Pharmacol Sin. 22, 1159-1167
  26. Marklund S., and Marklund G., (1974). Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem. 47, 469-474 https://doi.org/10.1111/j.1432-1033.1974.tb03714.x
  27. Misra H. P., (1974). Generation of superoxide free radical during the autoxidation of thiols. J Bioi Chem. 249, 2151-2155
  28. Nadkarni G. D., and D'Souza N. B., (1988). Antioxidant and free radical-scavenging enzymes in chronically ethanol-consuming rats: controversy over hepatic lipid peroxidation. Drug Alcohol Depend. 22, 161-164 https://doi.org/10.1016/0376-8716(88)90051-8
  29. Nordmann R., Ribiere C., and Rouach H., (1992). Implication of free radical mechanisms in ethanol-induced cellular injury. Free Radic Biol Med. 12, 219-240 https://doi.org/10.1016/0891-5849(92)90030-K
  30. Ohkawa H., Ohishi N., and Yagi K., (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 95, 351-358 https://doi.org/10.1016/0003-2697(79)90738-3
  31. Paglia D.E., and Valentine W. N., (1967). Glutathione peroxidase and selenoprotein activity in various tissue. J. Biol, Chem. 145, 233
  32. Pierson J. L, and Mitchell M. C., (1986). Increased hepatic efflux of glutathione after chronic ethanol feeding. Biochem Pharmacol. 35, 1533-1537 https://doi.org/10.1016/0006-2952(86)90121-8
  33. Recknagel R.O., GIende B.A., and Hruszkewyez A. M., (1977). Chemical mechanisms in carbon tetrachloride toxicity in 'Free radicals in biology'. Pryor, W. A. (ed.), Academic Press, New York, 97
  34. Reitz R. C., (1975). A possible mechanism for the peroxidation of lipids due to chronic ethanol ingestion. Biochim Biophys Acta. 380, 145-154 https://doi.org/10.1016/0005-2760(75)90001-6
  35. Ribiere C., Hininger I., and Rouach H., (1992). Effects of chronic ethanol administration on free radical defence in rat myocardium. Biochem Pharmacol. 44, 1495-1500 https://doi.org/10.1016/0006-2952(92)90463-S
  36. Rosenblum E. R., Gavaler J. S., and Van Thiel D. H., (1987). Vitamin A at pharmacologic doses ameliorates the membranelipid peroxidation injury and testicular atrophy that occurs with chronic alcohol feeding in rats. Alcohol and Alcoholism 22, 241-249
  37. Saravanan R., and Pugalendi V., (2006). Impact of ursolic acid on chronic ethanol-induced oxidative stress in the rat heart. Pharmacal Rep. 58, 41-47
  38. Schisler N. J., and Singh S. M., (1989). Effect of ethanol in vivo on enzymes which detoxify oxygen free radicals. Free Radic Biol Med. 7, 117-123 https://doi.org/10.1016/0891-5849(89)90002-6
  39. Schlorff E. C., Husain K., and Somani S. M., (1999). Dose- and time-dependent effects of ethanol on plasma antioxidant system in rat. Alcohol. 17, 97-105 https://doi.org/10.1016/S0741-8329(98)00039-1
  40. Sedlak J., and Lindsay R. H., (1968). Estimation of total, proteinbound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem. 25, 192-20 https://doi.org/10.1016/0003-2697(68)90092-4
  41. Sekura R. D., and Jakoby W. B., (1979). Phenol sulfotransferases. J Biol Chem. 254, 5658-5663
  42. Sherlock S., (1993). Disease of the Liver and Bilisry System. Oxford, Blackwell 8th, 370
  43. Thurman R. G., Bradford B. U., and Iimuro Y., (1997). Role of Kupffer cells, endotoxin and free radicals in hepatotoxicity due to prolonged alcohol consumption: studies in female and male rats. J Nutr. 127, 903S-906S https://doi.org/10.1093/jn/127.5.903S
  44. Vina J., Estrela J. M., and Guerri C., (1980). Effect of ethanol on glutathione concentration in isolated hepatocytes. Biochem J. 188, 549-552 https://doi.org/10.1042/bj1880549