Abstract
A hepatoprotective compound was isolated from the ethanolic extract of the fruits of Terminalia chebula Retz. by consecutive solvent partitioning, followed by silica gel and Sephadex LH-20 column chromatographies. The purified compound was identified as a mixture of chebulic acid and its minor isomer, neochebulic acid, with a ratio of 2:1 by spectroscopic analysis including 1D and 2D NMR and MS spectroscopy. To our knowledge, this is the first report on the protection of rat hepatocytes against oxidative toxicity by chebulic acid obtained from T. chebula Retz. This compound exhibited in vitro a free radical-scavenging activity and ferric-reducing antioxidant activity. Also, the specific ESR spectrum for the •OOH radical signals consisting of three-line ESR spectra was within the field of 0.27 mT, whereas 2.5 and 0.25 mg/ml of chebulic acid significantly reduced the signal intensity of the ESR spectra to 0.06 mT and 0.11 mT, respectively. Using isolated rat hepatocyte experiment, we demonstrated that the treatment of hepatocytes with chebulic acid significantly reduced the tert-butyl hydroperoxide (t-BHP)-induced cell cytotoxicity, intracellular reactive oxygen species level, and the ratio of GSSH, oxidized form of glutathione (GSH) to the over total GSH (GSH + GSSG) (4.42%) as compared to that with t-BHP alone (8.33%).
Similar content being viewed by others
References
Aqil F, Khan MS, Owais M, Ahmad I (2005) Effect of certain bioactive plant extracts on clinical isolates of beta-lactamase producing methicillin resistant Staphylococcus aureus. J Basic Microbiol 45:106–114
Arora S, Kaur K, Kaur S (2003) Indian medicinal plants as a reservoir of protective phytochemicals. Teratog Carcinog Mutagen 11:295–300
Asensi M, Sastre J, Pallardo FV., Garcia J, Estrela JM, Vina J (1994) A high-performance liquid chromatography method for measurement of oxidized glutathione in biological samples. Anal Biochem 217:323–328
Badmaev V, Nowakowski M (2000) Protection of epithelial cells against influenza a virus by a plant derived biological response modifier Ledretan-96. Phytother Res 14:245–249
Bajpai M, Pande A, Tewari SK, Prakash D (2005) Phenolic contents and antioxidant activity of some food and medicinal plants. Int J Food Sci Nutr 56:287–291
Beckman KB, Ames BN (1998) The free radical theory of aging matures. Physiol Rev 78:547–581
Berger MM (2005) Can oxidative damage be treated nutritionally? Clin Nutr 24:172–183
Bissell DM, Hammaker LE, Meyer UA (1973) Parenchymal celle from adult rat liver in nonproliferation monolayer culture: I. functional studies. J Cell Biol 59:722–734
Cheng HY, Lin TA, Yu KH, Yang CM, Lin CC (2003) Antioxidant and free radical scavenging activities of Terminalia chebula. Biol Pharm Bull 26:1331–1335
Chevalley I, Marston A, Hostettmann K (1999) A new gallic acid fructose ester from Saxifraga stellaris. Phytochemistry 50:151–154
Dalet C, Fehlmann M, Debey P (1982) Use of Percoll density gradient centrifugation for preparing isolated rat hepatocytes having long-term viability. Anal Biochem 122:119–123
Devasagayam TP, Tilak JC, Boloor KK, Sane KS, Ghaskadbi SS, Lele RD (2004) Free radicals and antioxidants in human health: current status and future prospects. J Assoc Physicians India 52:794–804
Freeman BA, Crapo JD (1982) Biology of disease: free radicals and tissue injury. Lab Invest 47:412–426
Gonzalez FJ (2005) Role of cytochromes P450 in chemical toxicity and oxidative stress: studies with CYP2E1. Mutat Res 569:101–110
Halliwell B, Gutteridge JM (1985) Oxygen radicals and the nervous system. Trends Neurosci 8:22–26
Jaeschke H (1990) Glutathione disulfide formation and oxidant stress during acetaminophen-induced hepatotoxicity in mice in vivo: the protective effect of allopurinol. J Pharmacol Exp Ther 255:935–941
Jiménez-Escrig A, Jiménez-Jiménez I, Pulido R, Saura-Calixto F (2001) Antioxidant activity of fresh and processed edible seaweeds. J Sci Food Agric 81:530–534
Juang LJ, Sheu SJ, Li TC (2004) Determination of hydrolyzable tannins in the fruit of Terminalia chebula Retz. by high-performance liquid chromatography and capillary electrophoresis. J Sep Sci 27:718–724
Kaur S, Grover IS, Singh M (1998) Antimutagenicity of hydrolyzable tannins from Terminalia chebula in Salmonella typhimurium. Mutat Res 419:169–179
Kitagaki H, Tsugawa M (1999) 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging ability of sake during storage. J Biosci Bioeng 87:328–332
Lee HS, Won NH, Kim KH, Lee H, Jun W, Lee KW (2005) Antioxidant effects of aqueous extract of Terminalia chebula in vivo and in vitro. Biol Pharm Bull 28:1639–1644
Naik GH, Priyadarsini KI, Naik DB, Gangabhagirathi R, Mohan H (2004) Studies on the aqueous extract of Terminalia chebula as a potent antioxidant and a probable radioprotector. Phytomedicine 11:530–538
Ochi T, Miyaura S (1989) Cytotoxicity of an organic hydroperoxide and cellular antioxidant defense system against hydroperoxides in cultured mammalian cells. Toxicology 55:69–82
Perry LM (1980) Medicinal plants of east and southeast Asia-attributed properties and use. The MIT, Cambridge, pp.80
Sabu MC, Kuttan R (2002) Anti-diabetic activity of medicinal plants and its relationship with their antioxidant property. J Ethnopharmacol 81:155–160
Saleem A, Husheem M, Härkönen P, Pihlaja K (2002) Inhibition of cancer cell growth by crude extract and the phenolics of Terminalia chebula Retz. fruit. J Ethnopharmacol 81:327–336
Skehan P, Storeng R, Scadiero D, Monks A, McMahon I, Vistica D, Warren JT, Bokesch H, Kenney S, Boy MR (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82:1107–1112
Tseng TH, Wang CJ, Kao ES, Chu HY (1996) Hibiscus protocatechuic acid protects against oxidative damage induced by tert-butylhydroperoxide in rat primary hepatocytes. Chem Biol Interact 101:137–148
Vonshak A, Barazani O, Sathiyamoorthy P, Shalev R, Vardy D, Golan-Goldhirsh A (2003) Screening South Indian medicinal plants for antifungal activity against cutaneous pathogens. Phytother Res 17:1123–1125
Wang H, Joseph JA (1999) Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic Biol Med 27:612–616
Wei T, Sun H, Zhao X, Hou J, Hou A, Zhao Q, Xin W (2002) Scavenging of reactive oxygen species and prevention of oxidative neuronal cell damage by a novel gallotannin, pistafolia A. Life Sci 70:1889–1899
Wickens AP (2001) Ageing and the free radical theory. Respir Physiol 128:379–391
Willcox JK, Ash SL, Catignan GL (2004) Antioxidants and prevention of chronic disease. Crit Rev Food Sci Nutr 44:275–295
Yagi K (1987) Lipid peroxides and human diseases. Chem Phys Lipids 45:337–351
Youdim KA, Joseph JA (2001) A possible emerging role of phytochemicals improving age-related neurological dysfunctions: a multiplicity of effects. Free Radic Biol Med 30:583–594
Acknowledgments
This work was supported by grant number 20050401-034-749-180-02-00 from the BioGreen 21 program, Rural Development Administration, Republic of Korea Research Fund (K0401771) in 2004–2005 and Ministry of Education and Human Resources Development in Korea (2005).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, HS., Jung, SH., Yun, BS. et al. Isolation of chebulic acid from Terminalia chebula Retz. and its antioxidant effect in isolated rat hepatocytes. Arch Toxicol 81, 211–218 (2007). https://doi.org/10.1007/s00204-006-0139-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-006-0139-4