文章摘要
张晓波,唐凤英,周 敏,朱 颖,陈 敏,周 惠,陈连华.IQGAP1在调节足细胞细胞骨架重组中的作用[J].南京医科大学学报,2017,(8):985~989
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
投稿时间:2016-07-13  
DOI:10.7655/NYDXBNS20170813
中文关键词: IQGAP1  足细胞  细胞骨架重组  嘌呤霉素
英文关键词: IQGAP1  podocytes  cytoskeleton rearrangement  puromycin aminonucleoside
基金项目:
作者单位
张晓波 南京医科大学附属淮安第一医院肾内科江苏 淮安 223300 
唐凤英 南京医科大学附属淮安第一医院肾内科江苏 淮安 223300 
周 敏 南京医科大学附属淮安第一医院肾内科江苏 淮安 223300 
朱 颖 南京医科大学附属淮安第一医院肾内科江苏 淮安 223300 
陈 敏 南京医科大学附属淮安第一医院肾内科江苏 淮安 223300 
周 惠 南京医科大学附属淮安第一医院肾内科江苏 淮安 223300 
陈连华 南京医科大学附属淮安第一医院肾内科江苏 淮安 223300 
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中文摘要:
      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IQGAP1 mRNA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;②IQGAP1 siRNA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pCantag-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。
英文摘要:
      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 mRNA 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:①when compared with the control group, the expression of IQGAP1 mRNA 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; ②the above rearrangement was exacerbated by IQGAP1 siRNA 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.
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