IQGAP1在调节足细胞细胞骨架重组中的作用  

The role of IQGAP1 in the regulation of cytoskeletal rearrangement in podocytes

作　　者：张晓波[1] 唐凤英[1] 周敏 朱颖[1] 陈敏[1] 周惠[1] 陈连华[1] 

机构地区：[1]南京医科大学附属淮安第一医院肾内科,江苏淮安223300

出　　处：《南京医科大学学报（自然科学版）》2017年第8期985-989,共5页Journal of Nanjing Medical University(Natural Sciences)

摘　　要：目的 :探讨IQ结构域GTP酶活化蛋白1(IQ domain GTPase-activating protein 1,IQGAP1)对足细胞细胞骨架重组的调节作用。方法:嘌呤霉素(PAN)刺激小鼠足细胞后,运用实时定量PCR和Western blot印迹法分别检测细胞内IQGAP1m RNA和蛋白表达水平,激光共聚焦显微镜观察足细胞骨架F-actin的分布,F-actin肌动蛋白评分系统(CFS)对骨架重组进行分析,并进一步观察上调和下调足细胞IQGAP1表达对PAN诱导的足细胞骨架重组的影响。结果:(1)与对照组相比,PAN刺激组足细胞内IQGAP1 mRNA和蛋白表达明显下降(P<0.05),足细胞F-actin排列紊乱,CFS明显升高;(2)IQGAP1 si RNA进一步抑制IQGAP1表达后,PAN引起的上述足细胞F-actin重组更加明显(P<0.05);(3)转染p Cantag-myc-IQGAP1质粒促进IQGAP1表达后,PAN诱导的足细胞F-actin排列紊乱显著减轻,CFS明显下降(P<0.05)。结论:IQGAP1能抑制嘌呤霉素诱导的足细胞骨架重组,对维持足细胞正常骨架结构起着重要作用。Objective： To investigate the regulatory role of IQGAP1 in cytoskeletal rearrangement of podocytes. Methods： Conditionally immortalized mouse podocytes were used to perform cell culture and stimulated by puromycin aminonucleoside（PAN）.RT-PCR and Western blot analysis were used to measure the level of IQGAP1 m RNA and protein,respectively. 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. We further explored the changes of cytoskeletal rearrangement after up-and down-regulation of IQGAP1. Results：（1）when compared with the control group, the expression of IQGAP1 m RNA and protein was significantly decreased after PAN stimulation in mouse podocytes（P〈0.05）. PAN stimulation induced cytoskeletal rearrangement including cortical F-actin ring formation, stress fiber attenuation and CFS up-regulation;（2） the above rearrangement was exacerbated by IQGAP1 si RNA transfection（P〈0.05）;（3） PAN induced cortical F-actin ring formation and CFS up-regulation was abated after IQGAP1 plasmid transfection（P〈0.05）. Conclusion： IQGAP1 can inhibit PAN induced cytoskeleton rearrangement in podocytes and plays an important role in maintaining normal podocyte cytoskeleton.

关 键 词：IQGAP1 足细胞 细胞骨架重组 嘌呤霉素 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]