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目的:研究支架蛋白IQGAP1在足细胞骨架重组中的作用。方法:用嘌呤霉素(PAN)刺激小鼠足细胞,实时定量PCR检测IQGAP1mRNA的表达,免疫印迹法检测足细胞IQGAP1蛋白的表达,激光共聚焦显微镜观察足细胞骨架F-actin分布,F-actin肌动蛋白评分系统(CFS)分析骨架重组;IQGAP1siRNA和IQGAP1质粒转染足细胞,分别观察下调和上调足细胞IQGAP1表达对嘌呤霉素诱导足细胞细胞骨架重组的改变。结果:嘌呤霉素刺激足细胞后IQGAP1mRNA和蛋白表达下调(P<0.05),足细胞F-actin分布发生明显重组;转染IQGAP1siRNA,嘌呤霉素刺激足细胞F-actin重组进一步加重;转染IQGAP1质粒,嘌呤霉素刺激足细胞F-actin重组减轻。结论:IQGAP1可能参与嘌呤霉素诱导的足细胞细胞骨架重组。
AIM: To investigate the role of scaffold protein IQGAP1 in podocyte cytoskeletal reorganization. Methods: The mouse podocytes were stimulated with puromycin (PAN). The expression of IQGAP1 mRNA was detected by real-time quantitative PCR. The expression of IQGAP1 protein in podocyte was detected by immunoblotting. The distribution of F-actin in podocyte cytoskeleton was observed by laser scanning confocal microscopy. Actin scoring system (CFS) analysis of skeletal reorganization; IQGAP1siRNA and IQGAP1 plasmid transfected podocytes, respectively, downregulation and upregulation of podocytes IQGAP1 expression induced by puromycin podocyte cytoskeletal reorganization. Results: The expression of IQGAP1mRNA and protein were down-regulated in puromycin-treated podocytes (P <0.05), and the distribution of F-actin in podocytes was significantly rearranged. IQGAP1siRNAs were transfected with puromycin to stimulate podocyte F- Plasmid, puromycin stimulated podocyte F-actin recombinant reduction. Conclusion: IQGAP1 may be involved in puromycin-induced podocyte cytoskeletal reorganization.