大鼠肾脏系膜细胞转染人TGF-β1基因可增强MMP-2表达  

Rat Mesangial Cell Transfected by Human Transforming Growth Factor β1 Gene Upregulates the Expression of Matrix Metalloproteinase 2

作　　者：林文生[1] 张农[1] 张颂文[2] 刘琛 顾建新[2] 郭慕依[1] 

机构地区：[1]上海医科大学基础医学院病理学教研室,上海200032 [2]上海医科大学基础医学院基因研究中心,上海200032

出　　处：《上海医科大学学报》2000年第1期13-15,27,共4页Journal of Fudan University(Medical Science)

基　　金：上海市科技发展基金!98QB14007;卫生部优秀青年科技人才专项基金;上海市教委重点学科基金!B980802

摘　　要：目的　研究转化生长因子β１（ＴＧＦβ１）对培养的大鼠肾小球系膜细胞（ＭｓＣ）基质金属蛋白酶２（ＭＭＰ２）表达的影响。方法　采用Ｌｉｐｏｆｅｃｔｉｎ将人ＴＧＦβ１基因转染培养的大鼠ＭｓＣ，经Ｎｏｒｔｈｅｒｎｂｌｏｔ鉴定其转染成功否；又分别应用Ｎｏｒｔｈｅｒｎｂｌｏｔ．Ｗｅｓｔｅｒｎｂｌｏｔ．酶谱分析法检测阳性表达人ＴＧＦβ１的ＭｓＣＭＭＰ２ｍＲＮＡ、蛋白及酶活性变化。结果　成功获得稳定过表达人ＴＧＦβ１的大鼠ＭｓＣ克隆（ＭＴＧ１），该细胞克隆的ＭＭＰ２ｍＲＮＡ．蛋白表达及其酶活性均获增强。结论　ＴＧＦβ１可增强ＭｓＣＭＭＰ２ｍＲＮＡ、蛋白表达，并提高其酶活性，这为进一步阐明ＴＧＦβ１在肾小球硬化发生机制中的作用提供了重要的实验手段和依据。Purpose To study the matrix metalloproteinase 2(MMP?2) expression on cultrured rat mesangial cells(MsC) transfected by human transforming growth factor β1(TGF?β1) gene. Methods Lipofectin method for transfecting human TGF?β1 gene into MsC;Northern blot analysis for detecting TGF?β1 mRNA level in MsC.MMP?2 mRNA,protein and enzyme activity were determined by Northern blot,Western blot,Zymography assay,respectively. Results The MsC clone (MTG1) overexpressed human TGF?β1 mRNA was successfully prepared and its expression of MMP?2 mRNA,protein and enzyme activity was increased. Results The MsC clone (MTG1) overexpressed human TGF?β1 mRNA was successfully prepared and its expression of MMP?2 mRNA,protein and enzyme activity was increased. Conclusions TGF?β1 might upregulate the expression of MMP?2 and enhance the enzymatic activity of MMP?2 on cultured MsC.It is valuable for explaining the role of TGF?β1 in the development of glomerulsclerosis.

关 键 词：大鼠 基因转染 TGFΒ1 MMP-2 肾小球疾病 

分 类 号：R692.602[医药卫生—泌尿科学]