丹参酚酸B对大鼠肝星状细胞中丝裂原激活的蛋白激酶通路的抑制作用  

Salianic-acid B inhibits MAPK signaling in activated rat hepatic stellate cells

作　　者：薛冬英[1] 洪嘉禾[1] 徐列明[1] 

机构地区：[1]上海中医药大学肝病研究所,200032

出　　处：《中华肝脏病杂志》2004年第8期471-474,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金(30271657);上海市教委重点学科建设项目

摘　　要：目的 探讨丹参酚酸B(SA-B)对活化的大鼠肝星状细胞(HSC)中丝裂原激活的蛋白激酶(MAPK)信号传导通路的抑制作用。 方法 分离大鼠HSC,于无包被塑料培养皿中原代培养7 d,继以10-6mol/L浓度的SA-B孵育,再加10 ng/ml的转化生长因子-β1(TGF-β1)刺激。以蛋白印迹法观察SA-B对TGF-β1刺激的HSC内细胞外调节的蛋白激酶(ERK)表达及其磷酸化和对HSC表面TGF-β1Ⅰ型受体(TβRⅠ)、Ⅱ型受体(TβpRⅡ)表达的影响,观察由此改变致HSC合成Ⅰ型胶原蛋白的变化。ELISA法测定HSC培养液中Ⅰ型胶原的含量;酶谱法观察基质金属蛋白酶2、9、13(MMP-2、MMP-9、MMP-13)的活性。 结果10-6mol/L浓度的SA-B可抑制原代正常培养9 d的HSC及TGF-β1刺激的HSC中ERK1/2的磷酸化,对TβRⅠ和TβRⅡ的表达无影响。SA-B可抑制原代正常培养的HSC合成Ⅰ型胶原,抑制TGF-β1刺激的HSC Ⅰ型胶原的合成及分泌。SA-B对HSC培养液中MMP-2和MMP-13的活性无明显作用,对MMP-9活性则有明显的促进作用。 结论 SA-B抑制了正常原代培养已活化的HSC和经TGF-β1刺激的HSC内ERK信号传导通路,这一抑制作用与HSC的TGF-β1受体表达无关。SA-B通过抑制TGF-β1的信号传导,减少了HSC对Ⅰ型胶原的合成与分泌,此种细胞功能的改变可能与基质金属蛋白酶的降解作用无关。Objective To investigate the inhibiting effect of salianic-acid B (SA-B) on mitogen-activated protein kinase (MAPK) Signaling in activated rat hepatic stellate cells (HSCs). Methods HSCs were isolated from normal rat by in situ perfusion and Nycodenz density-gradient centrifugation method. HSCs were primarily cultured on uncoated plastic for 7 days. Then cells were stimulated with 10 ng/ml transforming growth factor-β1 (TGF-β1) after incubated with 10-6M/L SA-B. The effects of SA-B on Extracellular-regulated kinase (ERK) expression and its phosphorylation. Transforming growth factor p, receptorⅠ(TβRⅠ) and transforming growth factorβ1 receptor Ⅱ(TβRⅡ) on HSCs, typeⅠcollagen expression in HSC Induced by TGF-β1 were detected with western blot assay. Quantity of TypeⅠcollagen in the medium of HSCs was detected by ELISA. Matrix metalloproteinase 2, 9, 13 (MMP-2, MMP-9 and MMP-13) in the medium of HSCs was tested by Zymography. Results The phosphorylation of ERK1/2 in HSCs with or without TGF-β1 was inhibited by SA-B. The expression of Tβ RⅠand TβRⅡon HSCs can not be affected by SA-B. The synthesization of Type Ⅰcollagen in HSCs was decreased by SA-B; The synthesization and secretion of typeⅠcollagen in HSCs with TGF-β1 were reduced by SA-B too. SA-B had no effect on the activity of MMP-2 and MMP-13, but induced the activity of MMP-13. Conclusions SA-B inhibits ERK signaling induced by TGF-β1 in HSC. This inhibition has no association with the expression of TβRⅠand TβRⅡon HSCs. SA-B reduces the synthesization and secretion of TypeⅠcollagen in HSC by means of inhibiting TGF-β1 signaling, which might be not related to the degrading activities of MMPs.

关 键 词：丹参酚酸B 大鼠 肝星状细胞 丝裂原激活的蛋白激酶 抑制作用 信号传导通路 肝纤维化 

分 类 号：R285[医药卫生—中药学]