PINCH-1-ILK-а-parvin复合物在足细胞中的表达及意义

Expression and significance of PINCH-1-ILK-а-parvin in podocyte

目的通过对条件永生性小鼠足细胞的研究,探讨PINCH-1-ILK-а-parvin(简称PIP复合物)在足细胞中的表达及意义。方法培养条件永生性小鼠足细胞,分为正常组、PINCH-1小干扰RNA(PINCH-1 siRNA)组、小干扰RNA阴性对照(siRNA)组。免疫沉淀法检测PIP复合物在足细胞中的表达,荧光显微镜下观察足细胞骨架F-actin的改变,光镜下观察足细胞黏附能力及足突伸展状态并测定伸展率和黏附率。结果转染PINCH-1 siRNA后,PINCH-1、ILK表达均下调:ILK/а-Parvin和PINCH-1/а-parvin在3组中的表达分别为normal组14.833±0.624、5.012±0.356;scrambled siRNA组11.333±1.886、4.511±0.816(P>0.05);PINCH-1siRNA组8.867±0.660、3.502±0.374(P<0.05)。转染PINCH-1 siRNA后,足细胞的伸展及黏附力均明显下降;足细胞伸展百分比分别为:normal组(68±4.243)%,PINCH-1 siRNA组(52±2.160)%,scrambled siRNA组(67.5±4.708)%(P>0.05);足细胞黏附率分别是:正常组(67±5.099)%,PINCH-1 siRNA组(50±5.888)%(P<0.05),scrambled siRNA组(66.5±5.212)%(P>0.05)。结论 PIP复合物通过介导足细胞骨架改变及黏附异常参与足细胞损伤的发生。

Objective To investigate the expression and possible roles of PINCH - 1 - ILK - a - parvin （PIP） complex in podocyte. Methods Cultured murine podocytes in vitro were pretreated with or without PINCH - 1 siRNA for variable concentrations and periods. The expression of PIP complex was detected by co - immunoprecipitation. And F - actin was analyzed by immunofluorescence and cell adhesion and spreading assays were measured as well. Results Knockdown of PIP complex protein （ using siRNA） resulted in reduced expression of PINCH - 1 and ILK,reorganization of cytoskeleton and decreasing of podocyte adhesion and spreading. Conclusion The PIP complex plays an important role in the development of podocyte injury through regulating the cytoskeleton and cell adhesion of podocytes.