Effects of salvianolic acid- B on TGF-beta1 stimulated hepatic stellate cell activation and its intracellular signaling

	Effects of salvianolic acid- B on TGF-beta1 stimulated hepatic stellate cell activation and its intracellular signaling
	Liu Chenghai; Liu Ping; Hu Yiyang; Zhu Dayuan
刊名	National Medical Journal of China
	2002
卷号	82 期号:18 页码:1267-1272
关键词	liver cirrhosis salvia miltiorhiza receptors transforming growth factor-beta signal transduction
ISSN号	0376-2491
其他题名	丹酚酸B盐对转化生长因子-beta1刺激肝星状细胞活化与胞内信号转导的作用
文献子类	Article
英文摘要	Objective To investigate hepatic stellate cells (HSC) responses at different differentiation stages on transforming growth factor-beta1, and to elucidate the mechanisms of salvianolic acid - B (SA-B) , a water soluble compound from Salvia miltiorrhiza , against hepatic fibrosis, relating to interference with TGF-(31 stimulated HSC activation and inlracellular signal transduction via Smads. Methods HSC was isolated from rat by in situ perfusion of liver and 8.2% nycondenz gradient centrifugation, and primarily cultured on uncoated plastic for 1 d, 4 d and 7 d respectively, which represented quiescent, intermediate and activated phenotypes. The cells were stimulated with 100 pmol/L TGF-pi, cell phenotypes were observed under inverted microscope, a-actin expression was checked by Western blot, and collagen secretion was measured with [ H ] proline incorporation and collangenase digestion, then HSC at one definite differentiation stage that responded most sensitively to TGF-J31 was selected as the cell model for the following study. 0.1 \|iimol/L~ 1 mmol/L SA-B was incubated with HSC and the cell proliferation was measured by intracellular [3H] thymidine pulse. SA-B was also incubated with TGF-pl stimulated HSC, the collagen secretion was measured as above, a-actin and plasmin activator inhibitor-1 (PAI-1) were checked with Western blot, and al(I) procollagen mRNA levels were analyzed with Northern blot. The cytoplasmie and nuclear proteins were extracted, and cytoplasmic and nuclear Smad2, 3 expression and phosphorylation levels were measured with Western blot.Results As culture duration prolonged, HSC phenotypes underwent activation gradually, accompanied by the increase of a-actin expression and collagen secretion. TGF-pi increased the basal collagen levels at dl, d4 and d7 by 128.6%, 207.7% and 188.2% of the control respectively, while d4 HSC had the most sensitive response, and this intermediate HSC was used as cell model for the following study. Except 0.1 mmol/L-1 mmol/L SA-B caused parts of HSC death, 0.1 /anol/L-10 (Mmol/L SA-B had no influence on cell shape, but decreased HSC proliferation in a dose depend manner, by 76% , 60.1% and 47.8% of the control respectively. 1 /imol/L-10 fxmol/L SA-B remarkably inhibited the collagen secretion of TGF-J31 stimulated HSC by 68.6% and 56.1% of the control, PAI-1 and a-actin expression, and down-regulated al(I) pro-collagen gene expression. 0.1 /umol/L - 10 jmmol/L SA-B decreased the cytoplasmic and nuclear Smad2,3 protein expression, especially inhibited Smad2 phosphorylation and nuclear translocation.Conclusions SA-B obviously inhibits intermediate HSC proliferation, decreases TGF-f!l stimulated HSC activation and matrix protein and gene expression, and inhibited stimulated HSC Smad2, 3 protein expression, phosphorylation and nuclear translocation. The inhibition of TGF-betal signaling in HSC and its biological responses is the important mechanism of SA-B against hepatic fibrosis.
资助项目	国家自然科学杰出青年基金资助项目（39825168）[00000000] ; 国家自然科学基金资助项目（39970899）[00000000]
WOS研究方向	General & Internal Medicine (provided by Clarivate Analytics)
语种	中文
CSCD记录号	CSCD:1089087
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/268474]
专题	中国科学院上海药物研究所
作者单位	Institute of Liver Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
推荐引用方式 GB/T 7714	Liu Chenghai,Liu Ping,Hu Yiyang,et al. Effects of salvianolic acid- B on TGF-beta1 stimulated hepatic stellate cell activation and its intracellular signaling[J]. National Medical Journal of China,2002,82(18):1267-1272.
APA	Liu Chenghai,Liu Ping,Hu Yiyang,&Zhu Dayuan.(2002).Effects of salvianolic acid- B on TGF-beta1 stimulated hepatic stellate cell activation and its intracellular signaling.National Medical Journal of China,82(18),1267-1272.
MLA	Liu Chenghai,et al."Effects of salvianolic acid- B on TGF-beta1 stimulated hepatic stellate cell activation and its intracellular signaling".National Medical Journal of China 82.18(2002):1267-1272.