Resveratrol effectively attenuates α-naphthyl-isothiocyanate-induced acute cholestasis and liver injury through choleretic and anti-inflammatory mechanisms
Abstract
Tao WANG1, 3, Zhi-xing ZHOU2, Li-xin SUN1, Xia LI1, Zhi-meng XU1, Mi CHEN1, Guo-lin ZHAO1, Zhen-zhou JIANG1, 3, *, Lu-yong ZHANG1, 4, *
1Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing 210009, China; 2Tianjin Key Laboratory of Molecular Design and Drug Discovery, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China; 3Jiangsu Center for Pharmacodynamics Research and Evaluation, China Pharmaceutical University, Nanjing 210009, China; 4State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China
Aim: α-Naphthylisothiocyanate (ANIT) is a well-characterized cholestatic agent for rats. The aim of this study was to examine whether resveratrol could attenuate ANIT-induced acute cholestasis and liver injury in rats.
Methods: SD rats were treated with resveratrol (15 or 30 mg/kg, ip) or a positive control drug ursodeoxycholic acid (100 mg/kg, po) for 5 consecutive days followed by a single dose of ANIT (60 mg/kg, po). Bile flow, and serum biochemical markers and bile constituents were measured 48 h after ANIT administration. Hepatic levels of oxidative repair enzymes (glutathione peroxidase, catalase and MnSOD), myeloperoxidase activity, TNF-α, IL-6, and ATP content, as well as the expression of liver transporter genes and proteins were assayed.
Results: ANIT exposure resulted in serious cholestasis and liver injury, as shown by marked neutrophil infiltration in liver, dramatically increased serum levels of ALT, AST, GGT, ALP, TBA, TBIL, IBIL, and DBIL, and significantly decreased bile excretion and biliary output of GSH and HCO3–. ANIT significantly increased TNF-α and IL-6 release and myeloperoxidase activity, decreased mitochondrial biogenesis in liver, but had little effect on hepatic oxidative repair enzymes and ATP content. Furthermore, ANIT significantly decreased the expression of Mrp2, FXR, and Cyp7a1, markedly increased Mrp3 expression in liver. Pretreatment with resveratrol attenuated ANIT-induced acute cholestasis and liver injury, and other pathological changes. Pretreatment with ursodeoxycholic acid was less effective.
Conclusion: Resveratrol effectively attenuates ANIT-induced acute cholestasis and liver injury in rats, possibly through suppression of neutrophil infiltration, as well as upregulation of expression of hepatic transporters and enzymes, thus decreasing accumulation of bile acids.
Keywords: resveratrol; ursodeoxycholic acid; α-naphthylisothiocyanate; cholestasis; liver injury; cytokine; bile acids; hepatic injury; cytokine; bile acids; hepatic transporters; FXR; Cyp7a1
This study was supported by grants from the National Natural Science Foundation of China (No 81320108029) and the 2011 Program for Excellent Scientific and Technological Innovation Team of Jiangsu Higher Education.
* To whom correspondence should be addressed.
E-mail lyzhang@cpu.edu.cn (Lu-yong ZHANG); jiangcpu@163.com (Zhen-zhou JIANG)
Received 2014-08-25 Accepted 2014-10-10
Keywords:
1Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing 210009, China; 2Tianjin Key Laboratory of Molecular Design and Drug Discovery, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China; 3Jiangsu Center for Pharmacodynamics Research and Evaluation, China Pharmaceutical University, Nanjing 210009, China; 4State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China
Aim: α-Naphthylisothiocyanate (ANIT) is a well-characterized cholestatic agent for rats. The aim of this study was to examine whether resveratrol could attenuate ANIT-induced acute cholestasis and liver injury in rats.
Methods: SD rats were treated with resveratrol (15 or 30 mg/kg, ip) or a positive control drug ursodeoxycholic acid (100 mg/kg, po) for 5 consecutive days followed by a single dose of ANIT (60 mg/kg, po). Bile flow, and serum biochemical markers and bile constituents were measured 48 h after ANIT administration. Hepatic levels of oxidative repair enzymes (glutathione peroxidase, catalase and MnSOD), myeloperoxidase activity, TNF-α, IL-6, and ATP content, as well as the expression of liver transporter genes and proteins were assayed.
Results: ANIT exposure resulted in serious cholestasis and liver injury, as shown by marked neutrophil infiltration in liver, dramatically increased serum levels of ALT, AST, GGT, ALP, TBA, TBIL, IBIL, and DBIL, and significantly decreased bile excretion and biliary output of GSH and HCO3–. ANIT significantly increased TNF-α and IL-6 release and myeloperoxidase activity, decreased mitochondrial biogenesis in liver, but had little effect on hepatic oxidative repair enzymes and ATP content. Furthermore, ANIT significantly decreased the expression of Mrp2, FXR, and Cyp7a1, markedly increased Mrp3 expression in liver. Pretreatment with resveratrol attenuated ANIT-induced acute cholestasis and liver injury, and other pathological changes. Pretreatment with ursodeoxycholic acid was less effective.
Conclusion: Resveratrol effectively attenuates ANIT-induced acute cholestasis and liver injury in rats, possibly through suppression of neutrophil infiltration, as well as upregulation of expression of hepatic transporters and enzymes, thus decreasing accumulation of bile acids.
Keywords: resveratrol; ursodeoxycholic acid; α-naphthylisothiocyanate; cholestasis; liver injury; cytokine; bile acids; hepatic injury; cytokine; bile acids; hepatic transporters; FXR; Cyp7a1
This study was supported by grants from the National Natural Science Foundation of China (No 81320108029) and the 2011 Program for Excellent Scientific and Technological Innovation Team of Jiangsu Higher Education.
* To whom correspondence should be addressed.
E-mail lyzhang@cpu.edu.cn (Lu-yong ZHANG); jiangcpu@163.com (Zhen-zhou JIANG)
Received 2014-08-25 Accepted 2014-10-10