Hepatoprotective effect of the ethanol extract of Vitis thunbergii on carbon tetrachloride-induced acute hepatotoxicity in rats through anti-oxidative activities Jeng-Shyan Deng a , Yi-Chih Chang b , Chi-Luan Wen c , Jung-Chun Liao d , Wen-Chi Hou e , Sakae Amagaya f , Shyh-Shyun Huang d,n , Guan-Jhong Huang g,nn a Department of Health and Nutrition Biotechnology, Asia University, Taichung 413, Taiwan b Department of Medical Laboratory Science and Biotechnology, China Medical University, Taiwan c Taiwan Seed Improvement and propagation Station, Council of Agriculture, Propagation Technology Section, Taichung, Taiwan d School of Pharmacy, China Medical University, Taichung 404, Taiwan e Graduate Institute of Pharmacognosy, Taipei Medical University, Taipei 250, Taiwan f Department of kampo Pharmaceutical Sciences, Nihon Pharmaceutical University, Saitama 362-0806, Japan g School of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung 404, Taiwan article info Article history: Received 21 December 2011 Received in revised form 21 May 2012 Accepted 2 June 2012 Available online 12 June 2012 Keywords: Vitis thunbergii Carbon tetrachloride Hepatotoxicity Nitric oxide Tumor necrosis factor-a abstract Ethnopharmacological relevance: Vitis thunbergii var. taiwaniana are traditionally used for the treatment of diarrhea, fracture and injury, jaundice, and hepatitis in Taiwan. Aim of the study: The hepatoprotective activity of its plant extracts seems to be been associated with its antioxidant activity. This paper aims to investigate the in vitro and in vivo antioxidant effects of the ethanol extract of Vitis thunbergii (EVT). Materials and methods: In HPLC analysis, the fingerprint chromatogram of EVT was established. Antioxidant ability of EVT was investigated by employing several established in vitro methods. In vivo antioxidant activity was tested against CCl 4 -induced toxicity in mice. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected in the blood to indicate hepatic injury. Product of lipid peroxidation (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH) contents were evaluated for oxidative stress in hepatic injury. Moreover, histopathological observation was assayed for the degree of hepatic injury. Results: EVT exhibited strong antioxidant ability in vitro. After oral administration of EVT significantly decreased ALT and AST, and ameliorated the oxidative stress in hepatic tissue and increased the activity of CAT, SOD, GPx, and GSH. Serum tumor necrosis factor-alpha (TNF-a), interleukin  1b (IL-1b), and nitric oxide (NO) were decreased in the group treated with CCl 4 plus EVT. Western blotting revealed that EVT blocked protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in CCl 4 -treated rats, significantly. Histopathological examination of livers showed that EVT reduced fatty degeneration, cytoplasmic vacuolization and necrosis in CCl 4 -treated rats. Conclusion: This study suggests that EVT possesses antioxidant effects in vitro and hepatoprotective effect on acute liver injuries induced by CCl 4 in vivo, and the results suggested that the effect of EVT against CCl 4 -induced liver damage is related to its antioxidant properties. & 2012 Elsevier Ireland Ltd. All rights reserved. 1. Introduction The oxidative damage caused by reactive oxygen species (ROS) and reactive nitrogen species (RNS) may generate various dis- eases in the human body, such as aging, arthritis, cancer, inflam- mation, heart diseases and other human diseases (Poli, 1993). The enhanced production of oxidative stress can be induced by a variety of factors, such as ionizing radiation, and exposure to drugs or xenobiotics (e.g., carbon tetrachloride). CCl 4 , an analogue of human hepatotoxin, has been used extensively in animal Contents lists available at SciVerse ScienceDirect journal homepage: www.elsevier.com/locate/jep Journal of Ethnopharmacology 0378-8741/$ - see front matter & 2012 Elsevier Ireland Ltd. All rights reserved. http://dx.doi.org/10.1016/j.jep.2012.06.003 Abbreviations: EVT, Vitis thunbergii; ALT, Alanine aminotransferase; AST, aspartate aminotransferase; SOD, superoxide dismutase; CAT, catalase; GPx, glutathione peroxidase; GSH, reduced glutathione; TNF-a, tumor necrosis factor-alpha; IL-1 b, interleukin-1b; NO, nitric oxide; iNOS, inducible NO synthase; COX-2, cyclooxygenase-2 n Corresponding author. nn Corresponding author. Tel.: þ886 4 2205 3366x5508. fax: þ886 4 2208 3362. E-mail addresses: wenhsin.press@msa.hinet.net (C.-L. Wen), gjhuang@mail.cmu.edu.tw (G.-J. Huang). Journal of Ethnopharmacology 142 (2012) 795–803