title: Sophocarpine attenuates liver fibrosis by inhibiting the TLR4 signaling pathway in rats.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  Sophocarpine can alleviate liver fibrosis mainly by inhibiting the TLR4 pathway. Sophocarpine may be a potential chemotherapeutic agent for chronic liver diseases.
authors.affiliation: 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.
authors.forename: Hui
authors.initials: H
authors.lastname: Qian
authors.affiliation: 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.
authors.forename: Jian
authors.initials: J
authors.lastname: Shi
authors.affiliation: 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.
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Fan
authors.affiliation: 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.
authors.forename: Jiao
authors.initials: J
authors.lastname: Lv
authors.affiliation: 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.
authors.forename: Si-Wen
authors.initials: SW
authors.lastname: Chen
authors.affiliation: 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.
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Song
authors.affiliation: 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.
authors.forename: Zhi-Wu
authors.initials: ZW
authors.lastname: Zheng
authors.affiliation: 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.
authors.forename: Wei-Fen
authors.initials: WF
authors.lastname: Xie
authors.affiliation: 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.
authors.forename: Yue-Xiang
authors.initials: YX
authors.lastname: Chen
mesh_terms: D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D001652:Bile Ducts / Q000601:surgery; D049109:Cell Proliferation; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D004396:Coloring Agents / Q000737:chemistry; D004128:Dimethylnitrosamine / Q000737:chemistry; D006909:Hydroxyproline / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008103:Liver Cirrhosis / Q000473:pathology*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction; D051197:Toll-Like Receptor 4 / Q000378:metabolism*; D000093842:Matrines
chemical_list: D000470:Alkaloids; D003094:Collagen; D004396:Coloring Agents; D004128:Dimethylnitrosamine; D006909:Hydroxyproline; D012333:RNA, Messenger; D051197:Toll-Like Receptor 4; C495347:Tlr4 protein, rat; C035933:sophocarpine; D000093842:Matrines
keywords: Cytokines; Hepatic stellate cells; Liver fibrosis; Sophocarpine; Toll-like receptor 4