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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Protective Effect of a 43 kD Protein from the Leaves of the Herb, Cajanus indicus L on Chloroform Induced Hepatic-disorder

  • Received : 2005.09.06
  • Accepted : 2006.01.12
  • Published : 2006.03.31
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Abstract

Cajanus indicus is a herb with medicinal properties and is traditionally used to treat various forms of liver disorders. Present study aimed to evaluate the effect of a 43 kD protein isolated from the leaves of this herb against chloroform induced hepatotoxicity. Male albino mice were intraperitoneally treated with 2mg/kg body weight of the protein for 5 days followed by oral application of chloroform (0.75ml/kg body weight) for 2 days. Different biochemical parameters related to physiology and pathophysiology of liver, such as, serum glutamate pyruvate transaminase and alkaline phosphatase were determined in the murine sera under various experimental conditions. Direct antioxidant role of the protein was also determined from its reaction with Diphenyl picryl hydraxyl radical, superoxide radical and hydrogen peroxide. To find out the mode of action of this protein against chloroform induced liver damage, levels of antioxidant enzymes catalase, superoxide dismutase and glutathione-S-transferase were measured from liver homogenates. Peroxidation of membrane lipids both in vivo and in vitro were also measured as malonaldialdehyde. Finally, histopathological analyses were done from liver sections of control, toxin treated and protein pre- and post-treated (along with the toxin) mice. Levels of serum glutamate pyruvate transaminase and alkaline phosphatase, which showed an elevation in chloroform induced hepatic damage, were brought down near to the normal levels with the protein pretreatment. On the contrary, the levels of anti-oxidant enzymes such as catalase, superoxide dismutase and glutathione-S-transferase that had gone down in mice orally fed with chloroform were significantly elevated in protein pretreated ones. Besides, chloroform induced lipid peroxidation was effectively reduced by protein treatment both in vivo and in vitro. In cell free system the protein effectively quenched diphenyl picryl hydrazyl radical and superoxide radical, though it could not catalyse the breakdown of hydrogen peroxide. Post treatment with the protein for 3 days after 2 days of chloroform administration showed similar results. Histopathological studies indicated that chloroform induced extensive tissue damage was less severe in the mice livers treated with the 43 kD protein prior and post to the toxin administration. Results from all these data suggest that the protein possesses both preventive and curative role against chloroform induced hepatotoxicity and probably acts by an anti-oxidative defense mechanism.

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References

  1. Abraham, P., Wilfred, G. and Cathrine, S. P. (1999) Oxidative damage to the lipids and proteins of the lungs, testis and kidney of rats during carbon tetrachloride intoxication. Clin. Chim. Acta 289, 177-179 https://doi.org/10.1016/S0009-8981(99)00140-0
  2. Achudume, A. C. (1991) Effects of DMSO on carbon tetrachloride induced hepatotoxicity in mice. Clin. Chim. Acta 200, 57-58 https://doi.org/10.1016/0009-8981(91)90335-A
  3. Adang, A. E., Brussee, J., van der Gen, A. and Mulder, G. J. (1990) The glutathione-binding site in glutathione Stransferases. Investigation of the cysteinyl, glycyl and gammaglutamyl domains. Biochem J. 269, 47-54 https://doi.org/10.1042/bj2690047
  4. Ansari, R. A., Tripathi, S. C., Patnaik, G. K. and Dhawan, B. N. (1991) Antihepatotoxic properties of Picroliv, an active fraction from rhizomes of Picorrhiza kurroa. J. Ethnopharmacol. 34, 61-68 https://doi.org/10.1016/0378-8741(91)90189-K
  5. Arena J. M. and Drew, R. H. (1986) Poisoning: Toxicology, Symptoms, Treatments. 5th ed., pp. 258-259, Charles C. Tomas Springfield IL
  6. Blois, M. S. (1958) Antioxidant determination by use of a stable free radical. Nature 29, 1199-1200
  7. Bonaventura, J., Schroeder, W. A. and Fang, S. (1972) Human erythrocyte catalase: an improved method of isolation and a revaluation of reported properties. Arch. Biochem. Biophys. 150, 606-617 https://doi.org/10.1016/0003-9861(72)90080-X
  8. 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
  9. Chopra, R. N., Nayar, S. L. and Chopra, I. C. (1986) Glossary of India's medicinal plants. Publication and Information Directorate, CSIR. New Delhi, 44
  10. Constan, A. A., Sprankle, C. S., Peters, J. M., Kedderis, G. L., Everitt, J. I., Wong, B. A., Gonzalez, F. L. and Butterworth, B. E. (1999) Metabolism of chloroform by Cyt P4502E1 is required for induction of toxicity in liver, kidney and nose of male mice. Toxicol. Appl. Pharmacol. 160, 120-126 https://doi.org/10.1006/taap.1999.8756
  11. Diez-Fernandez, C., Sanz, N. and Cascales, M. (1996) Changes in glucose-6-phosphate dehydrogenase and malic enzyme gene expression in acute hepatic injury induced by thioacetamide. Biochem. Pharmacol. 51, 1159-1163 https://doi.org/10.1016/0006-2952(96)00030-5
  12. Esterbauer, H. and Cheeseman, K. H. (1990) Determination of aldehydic lipid peroxidation products: Malonaldehyde and 4- hydroxynonenal. Methods Enzymol. 186, 407-421 https://doi.org/10.1016/0076-6879(90)86134-H
  13. Ghosh, A. and Biswas, K. (1973) Bhartiya Banausadhi (ed.) A. Chatterjee, Calcutta University Press, India. 2, 332-334
  14. Gokalp, O., Gulle, K., Sulak, O., Cicek, E. and Altuntos, I. (2003) The effects of methidathione on liver: role of vitamins E and C. Toxicol. Ind. Health 19, 63-67 https://doi.org/10.1191/0748233703th176oa
  15. Habig, W. H. and Jakoby, W. B. (1974) Glutathione STransferases. The first enzymatic step in mercapturic acid formation. J. Biol. Chem. 249, 7130-139
  16. Hayes, J. D. and Pulford, D. J. (1995) The glutathione S transferase supergene family: Regulation of glutathione S transferase and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. Critical Reviews Biochem. Mole. Biol. 30, 445-600 https://doi.org/10.3109/10409239509083491
  17. Kakkar, P., Das, B. and Viswanathan, P. N. (1984) A modified spectrophotometric assay of superoxide dismutase. Ind. J. Biochem. Biophys. 21, 130-132
  18. Kind, P. R. N. and King, E. J. (1954) Estimation of plasma phosphatase by determination of hydrolyzed phenol with antipyrine. J. Clin. Path. 7, 322-326 https://doi.org/10.1136/jcp.7.4.322
  19. Kirtikar, K. R. and Basu, B. D. (1935) Indian Medicinal Plants, Blatter, E., Caius, J. F., Mhaskarr, R. S. (eds.) pp. 809-811, Probasi Press, Calcutta, India
  20. Kluwe, W. M., McCormack, K. M. and Hook, J. B. (1978) Selective modification of the renal and the hepatic toxicities of chloroform by induction of drug-metabolizing enzyme systems in kidney and liver. J. Pharmacol. Exp. Ther. 207, 566-573
  21. Kurose, I., Higuchi, H., Miura, S., Saito, H., Watanabe, N., Hokari, R., Hirokawa, M., Takaishi, M., Zeki, S., Nakamura, T., Ebinuma, H., Kato, S. and Ishii, H. (1997) Oxidative stressmediated apoptosis of hepatocytes exposed to acute ethanol intoxication. Hepatology 25, 368-378 https://doi.org/10.1002/hep.510250219
  22. Kyle, M. E., Miccadei, S., Nakae, D. and Farber, J. L. (1987) Superoxide dismutase and catalase protect cultured hepatocytes from the cytotoxicity of acetaminophen. Biochem. Biophys. Res. Commun. 149, 889-896 https://doi.org/10.1016/0006-291X(87)90491-8
  23. Larson, J. L., Wolf, D. C. and Butterworth, B. E. (1993) Acute hepatotoxic and nephrotoxic effects of chloroform in male F- 344 rats and female B6C3F1 mice. Fundam. Appl. Toxicol. 20, 302-315 https://doi.org/10.1006/faat.1993.1040
  24. Lind, R. C., Begay, C. K. and Gandolfi, A. J. (2000) Hepatoprotection by dimethyl sulfoxide. III. Role of inhibition of the bioactivation and covalent bonding of chloroform. Toxicol. Appl. Pharmacol. 166, 145-150 https://doi.org/10.1006/taap.2000.8949
  25. Ljungman, A. G., Grum, C. M., Deeb, G. M., Bolling, S. F. and Morganroth, M. L. (1991) Inhibition of cyclooxygenase metabolite production attenuates ischemia-reperfusion lung injury. Am. Rev. Respir Dis. 143, 610-617 https://doi.org/10.1164/ajrccm/143.3.610
  26. Malhi, H., Irani, A. N., Gagandeep, S. G. and Gupta, S. (2002) Isolation of human progenitor liver epithelial cells with extensive replication capacity and differentiation into mature hepatocytes. J. Cell Sci. 115, 2679-2688
  27. Masuda, Y., Yano, I. and Murano, T. (1980) Comparative studies on the hepatotoxic action of chloroform and related halogenomathanes in normal and Phenobarbital pretreated animals. J. Pharmacobiodyn 3, 535-544
  28. Nishikimi, M., Rao, N. A. and Yagi, K. (1972) The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem. Biophys. Res. Commun. 46, 849-854 https://doi.org/10.1016/S0006-291X(72)80218-3
  29. Paoletti, F., Aldinucci, D., Mocalli, A. and Caparrini, A. (1986) A sensitive spectrophotometric method for the determination of superoxide dismutase activity in tissue extracts. Anal. Biochem. 154, 536-541 https://doi.org/10.1016/0003-2697(86)90026-6
  30. Reitman, S. and Frankel, S. A. (1957) Colorimetric method for the determination of serum glutamic oxaloacetic and glutamic pyruvic transminases. Am. J. Clin. Path. 28, 56-63 https://doi.org/10.1093/ajcp/28.1.56
  31. Sarkar, K., Ghosh, A. and Sil, P. C. (2005) Preventive and curative role of a 43 kD protein from the leaves of the herb Cajanus indicus on thioacetamide induced hepatotoxicity in vivo. Hepatol. Res. 33, 39-49 https://doi.org/10.1016/j.hepres.2005.06.007
  32. Smith, J. H., Maita, K., Sleight, S. D. and Hook, J. B. (1983) Mechanism of chloroform nephrotoxicity. I. Time course of chloroform toxicity in male and female mice. Toxicol. Appl. Pharmacol. 70, 467-479 https://doi.org/10.1016/0041-008X(83)90164-3
  33. Soni, M. G., Raniah, S. K., Mumtaz, M. M., Clewell, H. and Mehendale, H. M. (1999) Toxicant inflicted injury and stimulated tissue repair are opposing toxicodynamic forces in predictive toxicology. Regul. Pharmacol. Toxicol. 19, 165-174
  34. Stevens, J. L. and Anders, W. M. (1981) Effects of cysteine diethyl maleate and Phenobarbital treatments on the hepatotoxicity of [1H] and [2H] chloroform. Chem. Biol. Interact. 37, 207-217 https://doi.org/10.1016/0009-2797(81)90178-2
  35. Testai, E., Di Marzio, S., di Domenico, A., Piccardi, A. and Vittozzi, L. (1995) An in vitro investigation of the reductive metabolism of chloroform. Arch. Toxicol. 70, 83-88 https://doi.org/10.1007/BF02733667
  36. Thakore, K. N. and Mehendale, H. M. (1994) Effect of Phenobarbital and mirex pretreatments on carbon tetrachloride auto protection. Toxicol. Pathology 22, 291-299 https://doi.org/10.1177/019262339402200307
  37. Tomasi, A., Albano, E., Biasi, F., Slater, T. F., Vannini, V. and Dianzani, M. U. (1985) Activation of chloroform and related trihalomethanes to free radical intermediates in isolated hepatocytes and in the rat in vivo as detected by the ESR-spin trapping technique. Chem. Biol. Interact. 55, 303-316 https://doi.org/10.1016/S0009-2797(85)80137-X

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