丹参酚酸B盐对转化生长因子β1活化的大鼠肝星状细胞内细胞外信号调节激酶信号转导通路的抑制作用(英文)  

Savianolic acid B inhibits ERK signal transduction pathway activated by transforming growth factor-β1 in rat hepatic stellate cells

作　　者：宋雅芳[1,2,3] 吕志刚[1,2] 徐列明[1,2,4,5] 

机构地区：[1]上海中医药大学附属曙光医院肝硬化科,上海201203 [2]上海中医药大学肝病研究所,上海201203 [3]广州中医药大学脾胃研究所,广东广州510405 [4]肝肾疾病病证教育部重点实验室,上海201203 [5]上海市中医临床重点实验室,上海201203

出　　处：《中西医结合学报》2012年第4期454-461,共8页Journal of Chinese Integrative Medicine

基　　金：国家自然科学基金资助项目(No.30271657);上海市教育委员会重点学科建设项目(No.04CA02);上海市教育委员会重点学科(第五期)建设项目(No.J50307);上海市重点学科建设项目(No.Y0302);上海市高校E-研究院资助项目(No.E03008);上海市高校创新团队建设资助项目(第一期)

摘　　要：目的:探讨丹参酚酸B盐(salvianolic acid B,SA-B)对转化生长因子β1(transforming growthfactor-β1,TGF-β1)活化的大鼠肝星状细胞(hepatic stellate cell,HSC)内细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)信号转导通路的影响。方法:采用原位灌流酶消化及Nycodenz密度梯度离心的方法分离正常大鼠HSC,将TGF-β1和SA-B直接添加于原代HSC的无血清培养液中。蛋白印记法检测HSC内磷酸化和总ERK,以及有丝分裂原激活蛋白激酶激酶MEK、有丝分裂原活化蛋白激酶激酶激酶Raf、α平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和Ⅰ型胶原蛋白的表达。结果:SA-B可抑制正常原代HSC及TGF-β1刺激的HSC内ERK上游激酶MEK的磷酸化;而对正常原代HSC及TGF-β1刺激的HSC内MEK上游激酶Raf-1的磷酸化无明显抑制。SA-B对TGF-β1刺激的HSC内α-SMA表达有明显的抑制作用,SA-B与ERK阻断剂联用,几乎接近完全阻断α-SMA的表达。SA-B对正常培养的HSC和经TGF-β1刺激的HSCⅠ型胶原蛋白的合成都有明显的抑制作用。结论:SA-B抑制TGF-β1刺激的HSC内ERK信号转导通路的作用环节在于抑制了MEK的磷酸化。SA-B通过抑制TGF-β1的ERK信号转导通路,减少了因HSC活化导致的α-SMA表达和Ⅰ型胶原蛋白合成增加。OBJECTIVE： To investigate the effects of salvianolic acid B （SA-B） on extracellular signal-regulated kinase （ERK） signal transduction pathway activated by transforming growth factor-β1 （TGF-β1） in rat hepatic stellate cells （HSCs）. METHODS. HSCs were isolated from male Sprague-Dawley rats by in situ perfusion and Nycodenz density- gradient centrifugation method. TGF-β1 and SA-B were directly added to the serum-free medium of HSCs. Total and phosphorylated ERK, MEK, Raf and α-smooth muscle actin （α-SMA） and type 1 collagen were assayed by Western blotting. RESULTS. Phosphorylation of MEK in HSCs with or without TGF-β1 was inhibited by SA-B; however, phosphorylation of Raf in HSCs with or without TGF-~] was not inhibited by SA-B. Expression of α-SMA in HSCs with TGF-β1 was inhibited by SA-B. Combination of SA-B and the inhJbJtors of ERK （PD98059） can effectively inhibit the expression of α-SMA. SA-B also inhibited synthesization of type I collagen in HSCs with or without TGF-β1. CONCLUSION： The action point of SA-B inhibiting ERK signaling induced by TGF-β1 in HSCs is the inhibition of the phosphorylation of MEK. SA-B reduces the increase of expression of α-SMA and protein synthesization of type I collagen induced by TGF-β1 by means of inhibiting ERK signaling in activated HSCs of rats.

关 键 词：丹参酚酸B 肝星状细胞 转化生长因子Β1 细胞外信号调节MAP激酶类 大鼠 

分 类 号：R285.5[医药卫生—中药学]