摘要
目的 通过对大鼠肾小球系膜细胞 (MsC)转染反义转化生长因子 β1(TGF β1)基因 ,观察该细胞克隆基质金属蛋白酶家族 (MMPs)和纤溶酶原激活物抑制剂 1(PAI 1)表达的改变。方法 采用脂质体进行基因转染 ,并用Westernblot法加以鉴定 ,后分别应用Northernblot、Westernblot和酶谱分析法检测该细胞克隆MMPs和PAI 1表达的变化。结果 成功地建立低表达TGF β1的大鼠MsC克隆 (MAT1) ;该细胞MMP 2mRNA及蛋白表达增强 ,MMP 2 ,3 ,7酶活性也均获得提高 ,而PAI 1mRNA的表达则受到抑制。结论 从分子水平阻断MsCTGF β1的表达 ,大鼠肾小球系膜细胞MMPs表达增强和PAI 1表达降低。
Objective To study the plasminogen activator inhibitors 1(PAI-1) and matrix metalloproteinases (MMPs) expression on the cultured rat mesangial cells (MsC) transfected with antisense-TGF-β1 vector. Methods Lipofectin method was used to transfect antisense-TGF-β1 vector into MsC, Western blot and Northern blot analysis for detecting TGF-β1 peptides and antisense TGF-β1 RNA level. The expression of PAI-1 and MMPs were determined by Northern blot, Western bolt and zymography assay. Results The MsC clone (MAT1) which blocked the expression of TGF-β1 peptides was successfully established. The MsC clone showed increased expression of MMP-2 mRNA protein and enzyme activity of MMP-2, 3,7. However, expression of PAI-1 mRNA of MsC clone was suppressed. Conclusion Blocked expression of TGF-β1 can upregulate the expression of MMP-2,3,7 and downregulate the expression of PAI-1 mRNA in MsC. It is possible that antisense-TGF-β1 may be used for gene therapy in glomerular diseases.
出处
《中华病理学杂志》
CSCD
北大核心
2000年第4期284-287,共4页
Chinese Journal of Pathology
基金
上海市科技发展基金!资助项目 (98QB14 0 0 7)
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
关键词
肾小球膜
TGF-β1
MMPS
PAI-1
大鼠
Glomerular mesangium
Tranforming growth factor beta
RNA
antisense
Metalloproteases
Plasminogen activator inhibitors 1
