转化生长因子β1/Smad信号通路对大鼠肾系膜细胞基质金属蛋白酶2及其组织抑制因子2表达的影响  

Effect of transforming growth factor β1/Smad signaling pathway on the expression and enzymatic activity of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in cultured rat mesangial cells

作　　者：杨琛[1] 戴璐[1] 刘学光[1] 陈琦[1] 张秀荣[1] 郭慕依[1] 

机构地区：[1]复旦大学上海医学院病理学系,200032

出　　处：《中华病理学杂志》2003年第6期553-557,共5页Chinese Journal of Pathology

基　　金：上海市科委科学技术发展基金资助项目 (0 1JC14 0 18)

摘　　要：目的　探讨转化生长因子 β1(TGF β1) /Smad信号通路对大鼠肾系膜细胞 (MsC)基质金属蛋白酶 2 (MMP 2 )及其组织抑制因子 2 (TIMP 2 )蛋白表达和酶活性的影响。方法　采用脂质体介导法 ,分别将Smad 2 3 7基因瞬时转染体外培养的大鼠MsC ,并用免疫荧光 逆转录 聚合酶链反应 (RT PCR)和Western印迹法检测其转染成功与否 ;再分别用Western印迹 酶谱法或反式酶谱法 ,观察转基因MsC及其在TGF β1作用下MMP 2和TIMP 2蛋白表达和酶活性的改变。 结果　转染Smad 2基因的MsC ,MMP 2和TIMP 2蛋白表达和酶活性均略有升高 ,蛋白表达分别增强 1 4倍和 1 1倍 ;酶活性分别提高 1 4倍和 1 3倍 ,经TGF β1作用后有明显升高 ,蛋白表达分别增加 2 8倍和 3 0倍 (P <0 0 1) ;酶活性分别提高 3 0倍和 1 7倍 (P <0 0 1) ;转染Smad 3基因的MsC ,TIMP 2蛋白表达和酶活性略有升高 ,蛋白表达增强 1 2倍 ,酶活性提高 1 2倍 ,TGF β1作用后升高更为明显 ,蛋白表达增强 2 9倍 ,酶活性提高 1 8倍 ,而MMP 2蛋白表达和酶活性均无明显变化 ;转染Smad 7基因的MsC ,MMP 2和TIMP 2蛋白表达和酶活性均略有下降 ,蛋白表达均降低 1 2倍 ,酶活性均下降 1 2倍 ,在TGF β1作用下更为明显 ,蛋白表达分别?Objective To explore the effect of transforming growth factor (TGF) β1/ Smad signaling pathway on the expression and enzymatic activity of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in cultured rat mesangial cells (MsC). Methods Lipofectin method was used to transfect Smad 2, Smad 3 and Smad 7 vectors into MsC; and immunofluorescence, RT-PCR and Western blot analysis were used to detect their transfection efficiency. The expression and enzymatic activity of MMP-2 and TIMP-2 were determined by Western blot, zymography or reverse zymography assay. Results MsC transfected with Smad 2 gene showed slightly increased expression and enzymatic activity of both MMP-2 and TIMP-2, which was more obvious upon stimulation by TGF-β1. MsC transfected with Smad 3 gene showed a slight upregulation of TIMP-2 expression and its enzymatic activity, which was enhanced after TGF-β1 stimulation. There was however no change in MMP-2 expression and its enzymatic activity. On the other hand, MsC transfected with Smad 7 gene showed a decrease in MMP-2 and TIMP-2 expression and enzymatic activity, which was especially obvious after stimulation by TGF-β1. Conclusions TGF-β1/Smad signaling pathway may play an important role in the pathogenesis of glomerulosclerosis, probably via MMP-2 and TIMP-2 expression and the associated enzymatic activity.

关 键 词：转化生长因子Β1 信号通路 大鼠 肾系膜细胞 基质金属蛋白酶2 组织抑制因子2 脂质体介导法 肾小球硬化 

分 类 号：R692.6[医药卫生—泌尿科学]