title: Salvianolic acid B inhibits hepatic stellate cell activation through transforming growth factor beta-1 signal transduction pathway in vivo and in vitro.
abstract: Salvianolic acid B (Sal B) is a major water soluble component extracted from Radix Salviae miltiorrhizae, a traditional Chinese herb widely used for treating cardiovascular and hepatic diseases. Sal B has been reported to inhibit transforming growth factor (TGF)-β1-stimulated hepatic stellate cells (HSCs) activation and collagen type I expression. In this study, we further investigated the mechanisms of Sal B on liver fibrosis relating to TGF-β/Smads signalling pathway, especially to TGF-β1 receptors. Liver fibrosis model was induced by intraperitoneal injection of dimethylnitrosamine (DMN) for four weeks. Rats were randomly divided into three groups: normal, model, and Sal B groups. Rats in Sal B group were treated by oral administration of Sal B for four weeks from the first day of DMN exposure. Hydroxyproline (Hyp) content in liver tissue was assayed using Jamall's method and collagen deposition was visualized using Sirius red staining. HSCs were isolated from normal rats, and were cultured primarily in uncoated plastics. At day 4 after isolation, cells were stimulated with 2.5 ng/mL TGF-β1, and treated with 1 and 10 µmol/L Sal B and 10 µmol/L SB-431542 (TβR-I inhibitor) for 24 h, respectively. Cell proliferation was examined with 5-ethynyl-2'-deoxyuridine assay. The expressions of alpha smooth muscle actin (α-SMA) and Smad3 were assayed by immunofluorescent stain and Western blotting. The expression of TβR-I was analysed by Western blotting and real-time polymerase chain reaction. The activity of TβR-I kinase was measured by ADP-Glo kinase assay. The results showed that Sal B could inhibit collagen deposition and reduce Hyp content significantly, and decrease expressions of TGF-β1 and TβR-I in fibrotic liver in vivo. Also, Sal B decreased the expressions of α-SMA and TβR-I, inhibited Smad3 nuclear translocation and down-regulated TβR-I kinase activity in vitro. These findings suggested that Sal B could prevent HSCs activation through TGF-β signalling pathway, i.e. inhibiting TGF-β1 expression, activity of TβR-I kinase and Smads phosphorylation.
authors.affiliation: Institute of Liver Diseases, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
authors.forename: Yan-Yan
authors.initials: YY
authors.lastname: Tao
authors.forename: Qing-Lan
authors.initials: QL
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Shen
authors.forename: Wen-Wei
authors.initials: WW
authors.lastname: Fu
authors.forename: Cheng-Hai
authors.initials: CH
authors.lastname: Liu
mesh_terms: D000818:Animals; D001572:Benzofurans / Q000737:chemistry / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism; D004195:Disease Models, Animal; D055166:Hepatic Stellate Cells / Q000187:drug effects*; D006909:Hydroxyproline / Q000378:metabolism; D008103:Liver Cirrhosis / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology; D008297:Male; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*; D053630:Depsides
chemical_list: D001572:Benzofurans; D024042:Collagen Type I; D006909:Hydroxyproline; D051785:Smad Proteins; D053773:Transforming Growth Factor beta1; C076944:salvianolic acid B; D053630:Depsides
keywords: Salvianolic acid B; hepatic stellate cell; liver fibrosis; receptor; transforming growth factor beta-1