TGFβ1对转染Smad4/Smad7基因的大鼠系膜细胞Ⅰ、Ⅳ型胶原表达的影响  

Effect of TGFβ1 on the Expression of Type Ⅰ and Type Ⅳ Collagens in Cultured Rat Mesangial Cells Transfected with Smad4/Smad7 Vector

作　　者：杨琛[1] 戴璐[1] 黄瑾[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,上海200032

出　　处：《复旦学报（医学版）》2004年第2期115-118,F002,共5页Fudan University Journal of Medical Sciences

基　　金：上海市科委科学技术发展基金 ( 0 1JC14 0 18)资助项目

摘　　要：目的　探讨Smad4 /Smad7信号蛋白在转化生长因子β1(TGFβ1)作用下对大鼠系膜细胞 (MsC)Ⅰ、Ⅳ型胶原表达的影响。方法　经脂质体介导分别将Smad4 /Smad7基因瞬时转染体外培养的大鼠MsC ,并用免疫荧光、RT PCR和Westernblot法检测其转染成功与否 ;再用Westernblot法分别观察转基因MsC及其在TGFβ1作用下 ,其Ⅰ、Ⅳ型胶原蛋白表达的改变。结果　与转染空载体的MsC相比较 ,转染Smad4基因的MsC ,其Ⅰ、Ⅳ型胶原蛋白表达分别增强 1.2和 1.8倍 ,经TGFβ1作用后升高 1.8和 3.3倍 (P <0 .0 1) ;而转染Smad7基因的MsC ,其Ⅰ、Ⅳ型胶原蛋白表达分别下降 1.4和 1.9倍 ,经TGFβ1作用后则降低 1.7和 2 .4倍 (P <0 .0 1)。 结论　TGFβ1/Smad信号通路可从正反两方面调节MsCⅠ。Purpose To explore the effect of TGFβ1 on the expression of type Ⅰ collagen (Col Ⅰ) and type Ⅳ collagen (Col Ⅳ) in cultured rat mesangial cells (MsC) transfected with Smad4/Smad7 vector. Methods Lipofection method was used to transiently transfect Smad4/Smad7 vector into MsC,and immunofluorescence,RT-PCR and Western blot assay were used to detect their transfection efficiency,respectively.The expression of Col Ⅰ and Col Ⅳ was determined by Western blot analysis. Results MsC transfected with Smad4 vector showed 1.2 and 1.8 fold increase of Col Ⅰ and Col Ⅳ expression,and the increase of Col Ⅰ and Col Ⅳ expressions reach 1.8 and 3.3 fold respectively (P<0.01) while stimulated by TGFβ1.However,MsC transfected with Smad7 vector showed 1.4 and 1.9 fold decreases of both collagen expression,which decreased 1.7 and 2.4 fold after TGFβ1 stimulation ( P<0.01),compared with MsC transfected with pcDNA3 vector. Conclusions It is possible that TGFβ1/Smad signaling pathway affects the progression of gomerulosclerosis by up-regulation and/or down-regulating the syntheses of Col Ⅰ and Col Ⅳ of MsC.

关 键 词：TGFΒ1 转染 Smad4/Smad7基因 大鼠 系膜细胞 Ⅰ、Ⅳ型胶原 表达 

分 类 号：R364.3[医药卫生—病理学]