Hepatoprotective Effects of Paecilomyces tenuipes Against Carbon Tetrachloride-induced Toxicity in Primary Cultures of Adult Rat Hepatocytes

  • Published : 2007.12.31


Paecilomyces tenuipes (PT), one of the Ascomycetes family, has been used for medicinal purposes due to its broad pharmacological activities. The present study was undertaken to investigate the hepatoprotective effects of PT water extracts against $CCl_4$-induced hepatotoxicity in primary cultures of adult rat hepatocytes. When the extract of PT was directly added into the culture medium at 1, 2, and 5 mg/ml, the extracts not only reduce the $CCl_4$-induced elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase, and lipid peroxide, but also protect cultured hepatocytes from $CCl_4$-induced reduction of reduced glutathione, glutathione reductase, glutathione-S-transferase, glutathione peroxidase, catalase and superoxide dismutase. In addition, the effects of PT water extracts on cytochrome P450 enzymes were relatively marginal, indicating that the hepatoprotective effects of PT extract against $CCl_4$-induced toxicity might not be due to the inhibition of $CCl_4$ activation. In conclusion, the PT extracts were effective in protecting against $CCl_4$ induced hepatotoxicity in hepatocyte cultures, at least in part, by scavenging free radicals, and by modulating enzyme systems involved in cellular oxidative stress.



  1. Aebi, H. (1984). Catalase in vitro. Method. Enzymol., 105, 121-126
  2. Ames, B.N., Shigenaga, M.K. and Hagen, T.M. (1993). Oxidants, antioxidants, and the degenerative diseases of aging. Proc. Natl. Acad. Sci., U.S.A., 90, 7915-7922
  3. Borchers, A.T., Stern, J.S., Hackman, R.M., Keen, C.L. and Gershwin, M.E. (1999). Mushrooms, tumors, and immunity. Proc. Soc. Exp. Biol. Med., 221, 281-293
  4. Brattin, W.J., Glende, E.A. Jr. and Recknagel, R.O. (1985). Pathological mechanisms in carbon tetrachloride hepatohepatotoxicity. Free Rad. Biol. Med., 1, 27-38
  5. Brautbar, N. and Williams, J. 2nd. (2002). Industrial solvents and liver toxicity: risk assessment, risk factors and mechanisms. Int. J. Hyg. Env. Health, 205, 479-491
  6. Brent, J.A. and Rumack, B.H. (1993). Role of free radicals in toxic hepatic injury. II. Are free radicals the cause of toxininduced liver injury? J. Toxicol. Clin. Toxicol., 31, 173-196
  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. Comporti, M. (1985). Lipid peroxidation and cellular damage in toxic liver injury. Lab. Invest., 53, 599-623
  9. Dickins, M. and Peterson, R.E. (1980). Effects of a hormonesupplemented medium on cytochrome P-450 content and mono-oxygenase activities of rat hepatocytes in primary culture. Biochem. Pharmacol., 29, 1231-1238
  10. Eaton, J.W. (1991). Catalases and peroxidases and glutathione and hydrogen peroxide: mysteries of the bestiary. J. Lab. Clin. Med., 118, 3-4
  11. Fortson, W.C., Tedesco, F.J., Starnes, E.C. and Shaw, C.T. (1985). Marked elevation of serum transaminase activity associated with extrahepatic biliary tract disease. J. Clin. Gastroenterol., 7, 502-505
  12. Gonzalez, F.J. (1988). The molecular biology of cytochrome P450s. Pharmacol. Rev., 40, 243-288
  13. Guengerich, F.P., Kim, D.H. and Iwasaki, M. (1991). Role of human cytochrome P-450 IIE1 in the oxidation of many low molecular weight cancer suspects. Chem. Res. Toxicol., 4, 168-179
  14. Habig, W.H., Pabst, M.J. and Jakoby, W.B. (1974). Glutathione S-transferase. The first enzymatic step in mercapturic acid formation. J. Biol. Chem., 249, 7130-7139
  15. Halliwell, B., Aeschbach, R., Loliger, J. and Aruoma, O.I. (1995). The characterization of antioxidants. Food Chem. Toxicol., 33, 601-617
  16. Hatono, S., Jimenez, A. and Wargovich, M.J. (1996). Chemopreventive effects of S-allylcysteine and its relationship to the detoxification enzyme glutathione S-transferase. Carcinogenesis, 17, 1041-1044
  17. Hove, E.L. and Seibold, H.R. (1955). Liver necrosis and altered fat composition in vitamin E-deficient swine. J. Nutr., 56, 173-186
  18. Jeong, H.G. (1999). Inhibition of cytochrome P450 2E1 expression by oleanolic acid: hepatoprotective effects against carbon tetrachloride-induced hepatic injury. Toxicol. Lett., 105, 215-222
  19. Ketterer, B. (1988). Protective role of glutathione and glutathione transferases in mutagenesis and carcinogenesis. Mutat. Res., 202, 343-361
  20. Kiho, T., Hui, J., Yamane, A. and Ukai, S. (1993). Polysaccharides in fungi. XXXII. Hypoglycemic activity and chemical properties of a polysaccharide from the cultural mycelium of Cordyceps sinensis. Biol. Pharm. Bull., 16, 1291-1293
  21. Kim, N.D., Kwak, M.K. and Kim, S.G. (1997). Inhibition of cytochrome P450 2E1 expression by 2-(allylthio)pyrazine, a potential chemoprotective agent: hepatoprotective effects. Biochem. Pharmacol., 53, 261-269
  22. Koop, D.R. (1986). Hydroxylation of p-nitrophenol by rabbit ethanol-inducible cytochrome P-450 isozyme 3a. Mol. Pharmacol., 29, 399-404
  23. Koop, D.R. (1992). Oxidative and reductive metabolism by cytochrome P450 2E1. FASEB J., 6, 724-730
  24. Kuo, Y.C., Tsai, W.J., Shiao, M.S., Chen, C.F. and Lin, C.Y. (1996). Cordyceps sinensis as an immunomodulatory agent. Am. J. Chinese Med., 24, 111-125
  25. Lee, J.H., Cho, S.M., Song, K.S., Hong, N.D. and Yoo, I.D. (1996a). Characterization of carbohydrate-peptide linkage of acidic heteroglycopeptide with immuno-stimulating activity from mycelium of Phellinus linteus. Chem. Pharm. Bull., 44, 1093-1095
  26. Lee, S.S., Buters, J.T., Pineau, T., Fernandez-Salguero, P. and Gonzalez, F.J. (1996b). Role of CYP2E1 in the hepatotoxicity of acetaminophen. J. Biol. Chem., 271, 12063- 12067
  27. Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem., 193, 265-275
  28. 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
  29. McCay, P.B., Lai, E.K., Poyer, J.L., DuBose, C.M. and Janzen, E.G. (1984). Oxygen- and carbon-centered free radical formation during carbon tetrachloride metabolism. Observation of lipid radicals in vivo and in vitro. J. Biol. Chem., 259, 2135-2143
  30. Nam, K.S., Jo, Y.S., Kim, Y.H., Hyun, J.W. and Kim, H.W. (2001). Cytotoxic activities of acetoxyscirpenediol and ergosterol peroxide from Paecilomyces tenuipes. Life Sci., 69, 229-237
  31. Ohkawa, H., Ohishi, N. and Yagi, K. (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem., 95, 351-358
  32. Paglia, D.E. and Valentine, W.N. (1967). Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J. Lab. Clin. Med., 70, 158-169
  33. Poli, G. (1993). Liver damage due to free radicals. Brit. Med. Bull., 49, 604-620
  34. Rahman, I., Smith, C.A., Lawson, M.F., Harrison, D.J. and MacNee, W. (1996). Induction of ã-glutamylcysteine synthetase by cigarette smoke is associated with AP-1 in human alveolar epithelial cells. FEBS Lett., 396, 21-25
  35. Recknagel, R.O., Glende, E.A. Jr., Dolak, J.A. and Waller, R.L. (1989). Mechanisms of carbon tetrachloride toxicity. Pharmacol. Therapeut., 43, 139-154
  36. Recknagel, R.O., Glende, E.A. Jr. and Britton, R.S. (1991). Free radical damage and lipid peroxidation. In: Meeks, R.G., Harrison, S.D. and Bull, R.J. (Eds.), Hepatotoxicology. CRC Press, Florida, USA, pp. 401-436
  37. Salocks, C.B., Hsieh, D.P. and Byard, J.L. (1981). Butylated hydroxytoluene pretreatment protects against cytotoxicity and reduces covalent binding of aflatoxin B1 in primary hepatocyte cultures. Toxicol. Appl. Pharmacol., 59, 331- 345
  38. Sedlak, J. and Lindsay, R.H. (1968). Estimation of total, protein- bound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal. Biochem., 25, 192-205
  39. Stacey, N. and Priestly, B.G. (1978). Dose-dependent toxicity of $CCI_4$ in isolated rat hepatocytes and the effects of hepatoprotective treatments. Toxicol. Appl. Pharmacol., 45, 29-39
  40. von Herbay, A., Stahl, W., Niederau, C., von Laar, J., Strohmeyer, G. and Sies, H. (1996). Diminished plasma levels of vitamin E in patients with severe viral hepatitis. Free Rad. Res., 25, 461-466
  41. Williams, A.T. and Burk, R.F. (1990). Carbon tetrachloride hepatotoxicity: an example of free radical-mediated injury. Semin. Liver Dis., 10, 279-284
  42. Yang, K.H., Choi, E.J. and Choe, S.Y. (1983). Cytotoxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin on primary cultures of adult rat hepatocytes. Arch. Env. Contam. Toxicol., 12, 183-188
  43. Yoshikawa, T., Furukawa, Y., Murakami, M., Takemura, S. and Kondo, M. (1996). Effects of vitamin E on D-galactosamine- induced or carbon tetrachloride-induced hepatotoxicity. Digestion, 25, 222-229
  44. Zangar, R.C., Benson, J.M., Burnett, V.L. and Springer, D.L. (2000). Cytochrome P450 2E1 is the primary enzyme responsible for low-dose carbon tetrachloride metabolism in human liver microsomes. Chem. Biol. Interact., 125, 233-243