摘要
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。
Objective：To investigate the role of IQ-domain GTP-ase activated protein 1（IQGAP1）in cytoskeleton reorganization of podocytes.Methods：The mRNA and protein expression of IQGAP1 was detected by realtime-quantitative PCR and Western blotting in the puromycin aminonucleoside（PAN）-exposed podocytes.The cytoskeleton of F-actin was evaluated by FITC-conjugated phalloidin staining under laser confocal scanning microscope and semi-quantitative system with cortical F-actin score（CFS）was used to analyze the degree of actin cytoskeleton arrangement.IQGAP1 siRNAs and plasmid were introduced to decrease and overexpress the expression of IQGAP1 and the cytoskeleton reorganization was re-assessed in IQGAP1-regulated podocytes.Results：The expression of IQGAP1 mRNA and IQGAP1 protein was significantly decreased by PAN stimulation in mouse podocytes（P〈0.05）.PAN stimulation induced cytoskeletal rearrangement including cortical F-actin ring formation and stress fiber attenuation,which was exacerbated by IQGAP1 siRNA transfection and alleviated by IQGAP1 plasmid transfection in podocytes.Conclusion：IQGAP1may contribute to PAN-induced cytoskeleton reorganization in podocytes.
出处
《武汉大学学报（医学版）》
CAS
2015年第1期6-10,共5页
Medical Journal of Wuhan University
基金
国家自然科学基金资助项目(编号:81100478
81270762)
中央高校基本科研业务费专项资金(编号:111031)
