Effect of salvianolic acid B on collagen production and mitogen-activated protein kinase activity in rat hepatic stellate cells  

丹酚酸B对大鼠肝星状细胞胶原生成与丝裂原激活蛋白激酶活性的影响(英文)

作　　者：刘平 刘成海[1] 王海南[1] 胡义扬[1] 刘成[1] 

机构地区：[1]上海中医药大学肝病研究所,上海中国200032

出　　处：《Acta Pharmacologica Sinica》2002年第8期733-738,共6页中国药理学报（英文版）

基　　金：Project supported by the National Science Foundation for Outstanding Youth,№ 39585128.

摘　　要：AIM: To investigate the mechanism of salvianolic acid -B (SA-B) action against liver fibrosis relating to mediating hepatic stellate cell (HSC) activation and transforming growth factor-β1 (TGF-β1) intracellular signal transduction. METHODS: HSC was isolated from normal rat through in situ perfusion of liver with pronase E and density-gradient centrifugation with 11 % nycondenz, then cells were subcultured. Cell proliferation was observed by [3H]TdR uptake. Cellular collagen deposition was measured with Ponceau S stain and semi-quantified with image analytic system. Type I collagen secretion in the supernatant was detected with ELISA. The gene expression of type I pro-collagen was analyzed by RT-PCR. The supernatant was acidified and active TGF-β1 contents were assayed with ELISA. Mitogen-activated protein kinase (MAPK) activity was analyzed with immunoprecipitation and Western blot. RESULTS: SA-B 0.1, 1, 10, and 100 μmol/L suppressed HSC proliferation concentration-dependently as determined by [3H]TdR uptake by 94.1 %, 82.4 %, 62.7 %, and 4 % of the control respectively (P<0.05 or P0.01). SA-B 1, 10, and 100 μmol/L inhibited soluble type I collagen secretion by 75.3 %, 69.8 %, and 63.5 % of the control and decreased the matrix collagen deposition to 86.2 %, 75.4 %, and 73.4 % (P<0.05 or P< 0.01). SA-B 1 and 10 μmol/L decreased the cell active TGF-β1 secretion by 63.3 % and 15.6 % of the control, down-regulated pro-collgen α1(I) mRNA expression to 77.0 % and 51.8 % respectively (P<0.05). SA-B 1 and 10 μmol/L also inhibited MAPK activity by 1 to 2 fold respectively. CONCLUSION: SA-B inhibited HSC proliferation and collagen production as well as decreased the cells' TGF-β1 autocrine and MAPK activity, which might contri-bute to the mechanism of SA-B action against hepatic fibrosis.目的：研究丹酚酸B影响肝星状细胞活化与转化生长因子β1胞内 信号转导的抗肝纤维化作用机制。方法：正常大鼠肝脏链酶蛋白酶原位灌流消化与11％nycondenz密度梯度离心分离肝星状细胞，传一代培养。[^3H]TdR掺入法测定细胞增殖，丽春红染色、图像分析半定量细胞胶原沉积量，ELISA法测定细胞培养上清液I型胶原分泌量，培养上清酸化处理后，ELISA法测定活性TGF－β1含量。RT－PCR法分析细胞前胶原α1（Ⅰ）基因的表达，免疫沉淀一蛋白印迹法分析丝裂原激活蛋白激酶（MAPK）活性。结果：丹酚酸B0．1－100μmol/L浓度依赖性抑制性抑制细胞的I型胶原分泌量与总胶原的沉积。丹酚酸B1－10μmol/L抑制TGF－β1自分泌量，下调α1（Ⅰ）前胶原的基因表达，而且丹酚酸B1－10μmol/L明显抑制TGF－β1刺激的MAPK活性。结论：丹酚酸B可抑制肝星状细胞的增殖与胶原生成，抑制TGF－β1的自分泌与MAKP活性，这些作用是丹酚酸B抗肝纤维化的主要作用机制。

关 键 词：Salviae miltiorrhizae salvianolic acid B liver cirrhosis cell division COLLAGEN transforming growt factor beta mitogen-activated protein kinases signal transduction 

分 类 号：R96[医药卫生—药理学]