Abstract
Piper betel leaves (PBL) are used in Chinese folk medicine for the treatment of various disorders. PBL has the biological capabilities of detoxication, antioxidation, and antimutation. In this study, we evaluated the antihepatotoxic effect of PBL extract on the carbon tetrachloride (CCl4)-induced liver injury in a rat model. Fibrosis and hepatic damage, as reveled by histology and the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were induced in rats by an administration of CCl4 (8%, 1 ml/kg body weight) thrice a week for 4 weeks. PBL extract significantly inhibited the elevated AST and ALT activities caused by CCl4 intoxication. It also attenuated total glutathione S-transferase (GST) activity and GST α isoform activity, and on the other hand, enhanced superoxide dismutase (SOD) and catalase (CAT) activities. The histological examination showed the PBL extract protected liver from the damage induced by CCl4 by decreasing α-smooth muscle actin (α-sma) expression, inducing active matrix metalloproteinase-2 (MMP2) expression though Ras/Erk pathway, and inhibiting TIMP2 level that consequently attenuated the fibrosis of liver. The data of this study support a chemopreventive potential of PBL against liver fibrosis.
Similar content being viewed by others
References
Aimes RT, Quigley JP (1995) Matrix metalloproteinase-2 is an interstitial collagenase–Inhibitor-free enzyme catalyzes the cleavage of collagen. Brils and soluble native type I collagen generating the speci.c 3/4- and 1/4-length fragments. J Biol Chem 270:5872–5876
Allan JM, Wild CP, Rollinson S, Willett EV, Moorman AV, Dovey GJ, Roddam PL, Roman E, Cartwright RA, Morgan GJ (2001) Polymorphism in glutathione S-transferase P1 is associated with susceptibility to chemotherapy-induced leukemia. Proc Natl Acad Sci 98:11592–11597
Amonkar AJ, Nagabhushan M, D’Souza AV, Bhide SV (1986) Hydroxychavicol: a new phenolic antimutagen from betel leaf. Food Chem Toxicol 24:1321–1324
Amonkar AJ, Padma PR, Bhide SV (1989) Protection effect of hydroxychavicol, a phenolic component of betel leaf, against the tobacco-specific carcinogens. Mutat Res 210:249–253
Akiyoshi H, Terada T (1998) Mast cell, myofibroblast and nerve terminal complexes in carbon tetrachloride-induced cirrhotic rat livers. J Hepatol 29:112–119
Arthur MJ (2000) Fibrogenesis II. Metalloproteinases and their inhibitors in liver fibrosis. Am J Physiol Gastrointest Liver Physiol 279:G245–G249
Azuine MA, Bhide SV (1992) Protective single/combined treatment with betel leaf and turmeric against methyl (acetoxymethyl) nitrosamine-induced hamster oral carcinogenesis. Int J Cancer 51:412–415
Batist G, Tulpule A, Sinha BK, Katki AG, Myers CE, Cowan KH (1986) Overexpression of a novel anionic glutathione transferase in multidrug-resistant human breast cancer cells. J Biol Chem 261:15544–15549
Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287
Capdevielle-Pardies P, David J, Miquel JL, Le Bras M (1985) Quid of betel. Med Trop (Mar) 45:299–307
Chang MC, Uang BJ, Wu HL, Lee JJ, Hahn LJ, Jeng JH (2002) Inducing the cell cycle arrest and apoptosis of oral KB carcinoma cells by hydroxychavicol: roles of glutathione and reactive oxygen species. Br J Pharmacol 135:619–630
Choudhary D, Kale RK (2002) Antioxidant and non-toxic properties of Piper betle leaf extract: in vitro and in vivo studies. Phytother Res 16:461–466
Di Bisceglie AM (1997) Hepatitis C and hepatocellular carcinoma. Hepatology 26:34S–38S
Friedman SL (1997) Molecular mechanisms of hepatic fibrosis and principles of therapy. J Gastroenterol 32:424–430
Habig WH, Pabst MT, Jakoby WB (1974) Glutathione-S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139
Hengstler JG, Böttger T, Tanner B, Dietrich B, Henrich M, Knapstein PG, Junginger T, Oesch F (1998) Resistance factors in colon cancer tissue and the adjacent normal colon tissue: glutathione S-transferases C and J, glutathione and aldehyde dehydrogenase. Cancer Lett 128:105–112
Hernandez-Mounoz R, Diaz-Munoz M, Suarez-Cuenca JA, Trejo-Solis C, Lopez V, Sanchez-Sevilla L, Yanez L, De Sanchez VC (2001) Adenosine reverses a preestablished CCl4-induced micronodular cirrhosis through enhancing collagenolytic activity and stimulating hepatocyte cell proliferation in rats. Hepatology 34:677–687
Jeng JH, Kuo ML, Hahn LJ, Kuo MY (1994) Genotoxic and non-genotoxic effects of betel quid ingredients on oral mucosal fibroblasts in vitro. J Dent Res 73:1043–1049
Jeng JH, Wang YJ, Chang WH, Wu HL, Li CH, Uang BJ, Kang JJ, Lee JJ, Hahn LJ, Lin BR, Chang MC (2004) Reactive oxygen species are crucial for hydroxychavicol toxicity toward KB epithelial cells. Cell Mol Life Sci 61:83–96
Karaki Y, Munakata S, Saeki T, Hirota S, Fujimaki M (1991) Appearance of a carcinoid-like pattern in rat hepatic tumors induced by 3′-methyl-4-dimethyl-aminoazobenzene. Jpn J Cancer Res 82:397–402
Kitahara A, Satoh K, Nishimura K, Ishikawa T, Ruike K, Sato K, Tsuda H, Ito N (1984) Changes in molecular forms of rat hepatic glutathione S-transferase during chemical hepatocarcinogenesis. Cancer Res 44:2698–2703
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lear JT, Heagerty AHM, Smith A, Bowers B, Payne C, Smith CA, Jones PW, Gilford J, Yengi L, Alldersea J, Fryer AA, Strange RC (1996) Multiple cutaneous basal cell carcinomas: glutathione-S transferase (GSTM1, GSTT1) and cytochrome P-450 (CYP2D6, CYP1A1) polymorphisms influence tumour numbers and accrual. Carcinogenesis 17:1891–1896
Lee JY, Lee SH, Kim HJ, Ha JM, Lee SH, Lee JH, Ha BJ (2004) The preventive inhibition of chondroitin sulfate against the CCl4-induced oxidative stress of subcellular level. Arch Pharm Res 27:340–345
Lei D, Chan CP, Wang YJ, Wang TM, Lin BR, Huang CH, Lee JJ, Chen HM, Jeng JH, Chang MC (2003) Antioxidative and antiplatelet effects of aqueous inflorescence Piper betle extract. J Agric Food Chem 51:2083–2088
Lu FJ, Lin JT, Wang HP, Huang WC (1996) A simple, sensitive, non-stimulated photon counting system for detection of superoxide anion in whole blood. Experientia 52:141–144
McLellan LI, Judah DJ, Neal GE, Hayes JD (1994) Regulation of aflatoxin B1-metabolizing aldehyde reductase and glutathione S-transferase by chemoprotectors. Biochem J 300:117–124
Meyer DJ, Beale D, Tan KH, Coles B, Ketterer B (1985) Glutathione transferase in primary rat hepatomas: the isolation of a form with GSH peroxidase activity. FEBS Lett 184:139–143
Morrow CS, Smitherman PK, Diah SK, Schneider E, Townsend AJ (1998) Coordinated action of glutathione S-transferases (GSTs) and multidrug resistance protein 1 (MRP1) in antineoplastic drug detoxification. J Biol Chem 273:20114–20120
Murawaki Y, Yamada S, Ikuta Y, Kawasaki H (1999) Clinical usefulness of serum matrix metalloproteinase-2 concentration in patients with chronic viral liver disease. J Hepatol 30:1090–1098
Murphy G, Stanton H, Cowell S (1999) Mechanisms for pro matrix metalloproteinase activation. Acta Pathol Microbiol Immunol Scand 1:38–44
Nishida C, Ogawara KI, Kimura T, Higaki K (2004) Pharmacokinetic analysis of factors determining elimination pathways for sulfate and glucuronide metabolites of xenobiotics. III mechanisms for sinusoidal efflux of 4-methylumbelliferone sulfate. Xenobiotica 34:439–448
Norton SA (1998) Betel: consumption and consequences. J Am Acad Dermatol 38:81–88
Padma PR, Lalitha VS, Amonkar AJ, Bhide SV (1989) Anticarcinogenic effect of betel leaf extract against tobacco carcinogens. Cancer Lett 45:195–202
Paquet KJ, Kamphausen U (1975) The carbon-tetrachloride-hepatotoxicity as a model of liver damage. First report Long-time biochemical changes. Acta Hepatogastroenterol 22:84–88
Perez Tamayo R (1983) Is cirrhosis of the liver experimentally produced by CCl4 and adequate model of human cirrhosis? Hepatology 3:112–120
Rebbeck TR (1997) Molecular epidemiology of the human glutathione-S-transferase genotypes GSTM1 and GSTT1 in cancer susceptibility. Cancer Epidemiol Biomarkers Prev 6:733–743
Roeb E, Bosserhoff AK, Hamacher S, Jansen B, Dahmen J, Wagner S, Matern S (2005) Enhanced migration of tissue inhibitor of metalloproteinase overexpressing hepatoma cells is attributed to gelatinases: relevance to intracellular signaling pathways. World J Gastroenterol 11:1096–1104
Saravanan R, Rajendra Prasad N, Pugalendi KV (2003) Effect of Piper betel leaf extract on alcoholic toxicity in the rat brain. J Med Food 6:261–265
Seftor REB (1994) Electrophoretic analysis of proteins associated with tumor cell invasion. Electrophoresis 15:454–462
Sun Y, Elwell JH, Oberley LW (1988) A simultaneous visualization of the antioxidant enzymes glutathione peroxidase and catalase on polyacrylamide gels. Free Radic Res Commun 5:67–75
Takahara T, Furui K, Yata Y, Jin B, Zhang LP, Nambu S, Sato H, Seiki M, Watanabe A (1997) Dual expression of matrix metalloproteinase-2 and membrane-type 1-matrix metalloproteinase in fibrotic human livers. Hepatology 26:1521–1529
Watkins JB (1989) Exposure of rats to inhalational anesthetics alters the hepatobiliary clearance of cholephilic xenobiotics. J Pharmacol Exp Ther 250:421–427
Yang Y, Cheng JZ, Singhal SS, Saini M, Pandya U, Awasthi S, Awasthi YC (2001) Role of Glutathione S-Transferases in Protection against Lipid Peroxidation. J Biol Chem 276:19220–19230
Young SC, Wang CJ, Hsu JD, Hsu JL, Chou FP (2005) Increased sensitivity of Hep G2 cells toward the cytotoxicity of cisplatin by the treatment of piper betel leaf extract. Arch Toxicol 9:1–9
Acknowledgment
This work was supported by the grants from National Science Council (NSC94-2320-B-040-040) and Chung Shan Medical University (CSMC 92-OM-B-002), Taiwan.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Young, SC., Wang, CJ., Lin, JJ. et al. Protection effect of piper betel leaf extract against carbon tetrachloride-induced liver fibrosis in rats. Arch Toxicol 81, 45–55 (2007). https://doi.org/10.1007/s00204-006-0106-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-006-0106-0