Hepatoprotective Effect of Stamen Extracts of Mesua ferrea L. against Oxidative Stress induced by $CCl_4$ in Liver Slice Culture Model

  • Rajopadhye, Anagha A. (Botany group, Agharkar Research Institute) ;
  • Upadhye, Anuradha S. (Botany group, Agharkar Research Institute)
  • Received : 2011.09.09
  • Accepted : 2012.05.21
  • Published : 2012.06.30

Abstract

Stamens of Mesua ferrea L. are a well-known herbal drug used in Indian System of Traditional Medicine to treat various diseases. The claimed activity of this plant part is necessitated to investigate antioxidant and hepatoprotective activity. Authenticated plant sample was extracted with hexane, ethanol (EtOH) and water (aq.) using ASE 100 accelerated solvent extractor. Antioxidant activity was evaluated by means of different in vitro assays. Hepatoprotective effect was investigated on carbon tetrachloride induced oxidative stress in liver slice culture model. Cytotoxic marker lactate dehydrogenase (LDH) released in culture medium and the activity of lipid peroxidation along with antioxidant enzymes (AOEs) namely superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) were estimated. Hexane and EtOH extracts were significantly inhibited DPPH, NO, SOD and $ABTS^+$ radical in dose dependent manner. The trade of phenol content was: aq. extract < hexane extract < EtOH extract. A significant correlation was shown by total phenol content and free radical scavenging activity of extracts. The culture system treated with hexane extract, EtOH extract or ascorbic acid exhibited significant depletion in LDH, lipid peroxidation, antioxidative enzymes SOD, CAT and GR. Hexane extract and EtOH extracts of stamen of M. ferrea protected liver slice culture cells by alleviating oxidative stress induced damage to liver cells.

References

  1. Aebi, H.E., Catalase. In: Bergmeyer, H.U., (eds.), Methods of Enzymatic Analysis, Vol. 3, Verlagchemie GmbH, Weinhein, 1983, pp. 277-282
  2. Anonymous, The Wealth of India - A Dictionary of Indian Raw Material and Industrial Products, Raw Material Vol VI: L-M, New Delhi, CSIR, 1962.
  3. Bagul, M., Srinivasa, H., Anandjiwala, S. and Rajani, M., Phytochemical evaluation and free natural radical scavenging activity of stamen of Mesua ferrea Linn, var. ferrea. Indian Drugs 43, 665-670 (2006).
  4. Banerji, R. and Chowdhuary, A.R., Mesua ferrea: Chemical constituents and biological activity. J. Chem. Soc. Pak. 15, 207-211 (1993).
  5. Bradford, M.M., A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of dye binding. Anal. Biochem. 72, 248-254 (1976). https://doi.org/10.1016/0003-2697(76)90527-3
  6. Bulaj, G.T., Kortemme, T., and Goldenberg, D.P., Ionization-reactivity relationships for cysteine thiols in polypeptides. Biochem. 37, 8965-8972 (1998). https://doi.org/10.1021/bi973101r
  7. Desai, V.G., Aushadhi Sangraha, Shri Gajanan Book Depo, Bombay, 1975.
  8. Deshpande, A.P., Javalgekar, R.R., and Ranade, S., Dravyagunvidhyan, Anmola Prakhashan Pune, India, 1989.
  9. Dymock, W., Warden, C.J., and David, H., Pharmacographica Indica Vol I., Ballimaran Jayyed Press Delhi, 1890.
  10. Ellman, G.L., Tissue sulfhydryl groups. Arch. Biochem. Biophys. 82, 70-77 (1959). https://doi.org/10.1016/0003-9861(59)90090-6
  11. Frazier, J.M., In vitro toxicity testing: Application to safety evaluation, Marcel Dekker, Inc New York, 1992.
  12. Garg, S., Sharma, K., Ranjan, R., Attri, P., and Mishra, P., In vivo Antioxidant activity and hepatoprotective effects of methanolic extract of Mesua ferrea Linn. Int. J. PharmTech Res. 1, 1692-1696 (2009).
  13. Govindrajan, R., Vijayakumar, M., Rao, C.V., Shirwaikar, A., Rawat, A.K.S., Mehrotra, S., and Pushpangadan, P., Antioxidant potential of Anogeissus latifolia. Biol. Pharm. Bull. 7, 1266-1269 (2004).
  14. Hall, C.A. and Cuppett, S.L., Structure-activities of natural antioxidants, In: Aruoma, O.I. and Cuppett, S.L., (eds.), Antioxidant methodology: In vivo and in vitro concepts, AOCS Press Champaign, IL, 1997, pp. 141-172.
  15. Henry, L.E.A., Halliwell, B., and Hall, D.O., The Superoxide dismutase activity of various photosynthetic organisms measured by a new and rapid assay technique. FEBS Lett. 66, 303-306 (1976). https://doi.org/10.1016/0014-5793(76)80527-3
  16. Invittox Protocol No. 42., Liver slice hepatotoxicity screening system. The ERGATT/FRAME Data Bank of in vitro techniques in toxicology, INVITTOX, England, 1992.
  17. Joseph, C.R., Ilanchezhian, R., Biswajyoti, P., and Harish, C.R., Pharmacognostic studies of Nagakeshar (Mesua Ferrea L.) - An ingredient in Vyaghrihareetaki Avaleha. Int. J. Res. Ayurveda Pharm. 1, 264-272 (2010).
  18. Jung, K.-A., Song, T.-C., Han, D., Kim, I.-H., Kim, Y.-E., and Lee, C.-H., Cardiovascular protective Properties of Kiwi fruit extracts in vitro. Biol. Pharm. Bull. 28, 1782-1785 (2005). https://doi.org/10.1248/bpb.28.1782
  19. Magalhaes, L.M., Segundo, M.A., Reis, S., and Lima, L.M.C., Methodological aspects about in vitro evaluation of antioxidant properties. Anal. Chim. Acta. 61, 1-19 (2008).
  20. Makker, H.P.S., Dawra, R.K., and Singh, B., Determination of both tannin and protein in tannin-protein complex. J. Agric. Food Chem. 36, 523-525 (1988). https://doi.org/10.1021/jf00081a600
  21. Marcocci, L., Maguire, J.J., Droy-Lefaix, M.T., and Parker, L., The nitric oxide-scavenging properties of Ginkgo biloba extract EGB 761. Biochem. Biophys. Res. Comm. 201, 748-755 (1994). https://doi.org/10.1006/bbrc.1994.1764
  22. Marklund, S. and Marklund, G., Involvement of superoxide anion radical in auto oxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur. J. Biochem. 47, 469-464 (1974). https://doi.org/10.1111/j.1432-1033.1974.tb03714.x
  23. McMillan, D.C., Charles, B.J. and Jollow, D.J., Role of lipid peroxidation in dapsone-induced hemolytic anemia. J. Pharmacol. Exp. Ther. 287, 868-876 (1998).
  24. Miller, N.J., Diplock, A.T., and Rice-Evans, C.A., Evaluation of the total antioxidant as a marker of the deterioration of apple juice on storage. J. Agric. Food Chem. 43, 1794-1801 (1995). https://doi.org/10.1021/jf00055a009
  25. Naik, R.S., Mujumdar, A.M., and Ghaskadbi, S., Protection of liver cells from ethanol cytotoxicity by curcumin in liver slice culture in vitro. J. Ethnopharmacol. 95, 31-37 (2004). https://doi.org/10.1016/j.jep.2004.06.032
  26. Renner, K., Amberger, A., Konwalinka, G., Kofler, R., and Gnaiger, E., Changes of mitochondrial respiration, mitochondrial content and cell size after induction of apoptosis in leukemia cells. Biochem. Biophys. Acta. 1642, 115-123 (2003). https://doi.org/10.1016/S0167-4889(03)00105-8
  27. Sharma, P.C., Yelne, M.B., and Dennis, T.J., Database on medicinal plants used in Ayurveda Vol 5, CCRS, Department of ISM & H, Ministry of Welfare, India, 2002.
  28. Slinkard, K. and Singleton, V.L., Total phenol analysis: Automation and comparison with manual methods. Am. J. Enol. Vitic. 28, 49-55 (1977).
  29. Wormser, U. and Ben, Z.S., The liver slice system-an in vitro acute toxicity test for assessment of hepatotoxins and their antidotes. Toxicol. In Vitro 4, 449-451 (1990). https://doi.org/10.1016/0887-2333(90)90098-E