title: Tetramethylpyrazine inhibits CTGF and Smad2/3 expression and proliferation of hepatic stellate cells.
abstract: To study the effects of tetramethylpyrazine (TMP) on the proliferation of hepatic stellate cells-T6 (HSC-T6), and the expression of connective tissue growth factor (CTGF) and Smad2/3 in these cells, HSC-T6 cells were cultured with TMP at different concentrations after transforming growth factor-β1 (TGF-β1) stimulation. MTT assay was used to assess the cell proliferation. Cells were divided into the control group, TGF-β1-treated group and TMP-treated groups, which were treated with different concentrations of TMP. Immunocytochemistry and western blot were performed to detect the expression levels of CTGF and Smad2/3 in HSC-T6 cells. MTT analysis indicated that TMP significantly inhibited the proliferation of HSC-T6 cells, in dose-dependent and time-dependent manners. Immunocytochemistry detection and western blot showed that TMP could diminish TGF-β1-induced CTGF over-expression in HSC-T6 cells. Similarly, the enhancing effects of TGF-β1 on Smad2/3 expressions in HSC-T6 cells could also be counteracted by TMP treatment. Nuclear translocation of Smad2/3 was blocked by TMP treatment. Correlation analysis suggested a positive correlation between CTGF and Smad2/3 expression levels in HSC-T6 cells. TMP exerts anti-hepatic fibrosis effect through decreasing the expression of CTGF and Smad2/3, as well as inhibiting the proliferation of HSC-T6 cells. Our study provides cellular and molecular bases for further application of TMP in the clinical treatment for hepatic fibrosis.
authors.affiliation: Department of Endodontics, The Affiliated Hospital of Stomatology, Chongqing Medical University , Chongqing 400010 , China ; Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences , Chongqing 401147 , China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Endodontics, The Affiliated Hospital of Stomatology, Chongqing Medical University , Chongqing 400010 , China ; Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences , Chongqing 401147 , China.
authors.forename: Ni
authors.initials: N
authors.lastname: Dong
authors.affiliation: Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010 , China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Cheng
authors.affiliation: Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010 , China.
authors.forename: Xiaosheng
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010 , China.
authors.forename: Wenli
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University , Chongqing 400010 , China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xiang
keywords: Smad; TGF-β1; connective tissue growth factor; hepatic stellate cells; tetramethylpyrazine