Original Article
Copyright ©2014 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Feb 21, 2014; 20(7): 1822-1832
Published online Feb 21, 2014. doi: 10.3748/wjg.v20.i7.1822
Sophocarpine attenuates liver fibrosis by inhibiting the TLR4 signaling pathway in rats
Hui Qian, Jian Shi, Ting-Ting Fan, Jiao Lv, Si-Wen Chen, Chun-Yan Song, Zhi-Wu Zheng, Wei-Fen Xie, Yue-Xiang Chen
Hui Qian, Jian Shi, Ting-Ting Fan, Si-Wen Chen, Chun-Yan Song, Wei-Fen Xie, Yue-Xiang Chen, Department of Gastroenterology, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Hui Qian, Jiao Lv, Zhi-Wu Zheng, Department of Gastroenterology, the 411th Hospital of PLA, Shanghai 200081, China
Author contributions: Qian H, Shi J and Fan TT contributed equally to this study who performed the majority of the experiments and prepared the manuscript with assistance from Chen SW and Song CY; Lv J and Zheng ZW performed the immunohistochemical staining and data analysis; Xie WF designed the experiments and supervised the project; Chen YX designed the experiments, analyzed the data and wrote the manuscript.
Supported by The National Natural Science Foundation of China, Nos. 30971343, 81270486, 81000167 and 81370009
Correspondence to: Yue-Xiang Chen, Professor, Department of Gastroenterology, Shanghai Changzheng Hospital, Second Military Medical University, 415 Fengyang Rd, Shanghai 200003, China. yuexchen1807@163.com
Telephone: +86-21-81885343 Fax: +86-21-81886824
Received: September 7, 2013
Revised: November 13, 2013
Accepted: November 28, 2013
Published online: February 21, 2014
Processing time: 185 Days and 21.8 Hours
Abstract

AIM: To explore the effect of sophocarpine on experimental liver fibrosis and the potential mechanism involved.

METHODS: Sophocarpine was injected intraperitoneally in two distinct rat hepatic fibrosis models induced either by dimethylnitrosamine or bile duct ligation. Masson’s trichrome staining, Sirius red staining and hepatic hydroxyproline level were used for collagen determination. Primary hepatic stellate cells (HSCs) were isolated and treated with different concentrations of sophocarpine. Real-time reverse transcription-polymerase chain reaction was used to detect the mRNA levels of fibrotic markers and cytokines. The expression of pathway proteins was measured by Western blot. The Cell Counting Kit-8 test was used to detect the proliferation rate of activated HSCs treated with a gradient concentration of sophocarpine.

RESULTS: Sophocarpine decreased serum levels of aminotransferases and total bilirubin in rats under chronic insult. Moreover, administration of sophocarpine suppressed extracellular matrix deposition and prevented the development of hepatic fibrosis. Furthermore, sophocarpine inhibited the expression of α-smooth muscle actin (SMA), interleukin (IL)-6, transforming growth factor-β1 (TGF-β1), Toll-like receptor 4 (TLR4), and extracellular-related kinase (ERK) in rats. Sophocarpine also down-regulated the mRNA expression of α-SMA, collagen I, collagen III, TGF-β1, IL-6, tumor necrosis factor-α and monocyte chemoattractant protein-1, and decreased protein levels of TLR4, p-ERK, p-JNK, p-P38 and p-IKK in vitro after Lipopolysaccharide induction. In addition, sophocarpine inhibited the proliferation of HSCs accompanied by a decrease in the expression of Cyclin D1. The protein level of proliferating cell nuclear antigen was decreased in activated HSCs following a gradient concentration of sophocarpine.

CONCLUSION: Sophocarpine can alleviate liver fibrosis mainly by inhibiting the TLR4 pathway. Sophocarpine may be a potential chemotherapeutic agent for chronic liver diseases.

Keywords: Liver fibrosis; Sophocarpine; Toll-like receptor 4; Hepatic stellate cells; Cytokines

Core tip: Sophocarpine significantly ameliorated liver function and hepatic fibrosis in both the dimethylnitrosamine and bile duct ligation models. In addition, sophocarpine inhibited the activation and proliferation of hepatic stellate cells in vitro, which contributed to the anti-fibrotic effect of sophocarpine in vivo. Toll-like receptor 4 signaling was blocked by sophocarpine in vivo and in vitro, accompanied by a reduction in pro-inflammatory and fibrotic cytokines, as well a decrease in the expression of Cyclin D1 and proliferating cell nuclear antigen.