Exocyst敲减对肾足细胞缺失及功能紊乱的影响  

Effects of Exocyst knockdown on renal podocyte loss and dysfunction

作　　者：许晓丽 何志婷 

机构地区：[1]西安市第九医院肾脏内科

出　　处：《陕西医学杂志》2020年第1期3-6,15,共5页Shaanxi Medical Journal

基　　金：陕西省重点研发计划项目(2018SF-211)

摘　　要：目的:探讨Exocyst敲减对肾足细胞缺失及功能紊乱的影响。方法:采用对数生长期的人肾足细胞,分别转染对照组和shRNA Exoc5。流式检测细胞周期和凋亡,RT-PCR检测凋亡相关蛋白Caspase-3、Exoc1-8、裂孔膜蛋白Nephrin和肾小球蛋白Neph1的表达情况,免疫荧光检测细胞中裂孔膜蛋白Nephrin和Neph1定位情况,活细胞显微镜观察囊泡运动。结果:与对照组相比,Exocyst敲减肾足细胞细胞凋亡率明显升高、Caspase-3表达增加(P<0. 05);此外,Exocyst敲减导致Nephrin和Neph1错误定位和囊泡运动减少,过表达Exoc5可挽救Nephrin和Neph1定位。结论:本研究显示,Exocyst家族重要组分Exoc5敲减可导致肾足细胞凋亡增加,促使Nephrin和Neph1错误定位和囊泡运动减少,可能是肾足细胞损伤和功能紊乱的重要原因。Objective:To explore the effects of Exocyst knockdown on renal podocyte loss and dysfunction. Methods:The control group and shRNA Exoc5 were transfected with human renal podocytes in logarithmic growth phase. Cell cycle and apoptosis were detected by flow cytometry. The expression of apoptosis-related protein caspase-3,Exoc1-8,hiatal membrane protein Nephrin and Neph1 were detected by RT-PCR. The localization of Nephrin and Neph1 was detected by immunofluorescence. The vesicular movement was observed by living cell microscopy. Results:Compared with the control group,the apoptosis rate of Exocyst knockdown renal podocytes and the expression of caspase-3 were increased(P<0.05). In addition,the knockdown of Exocyst resulted in false localization of Nephrin and Neph1 and the decrease of vesicle movement. Over-expression of Exoc5 could save the localization of Nephrin and Neph1. Conclusion:The knockdown of Exoc5,an important component of the Exocyst family,can lead to the increase of renal podocyte apoptosis,the false localization of Nephrin and Neph1,and the decrease of vesicle movement,which may be an important cause of the injury and dysfunction of renal podocyte.

关 键 词：EXOCYST Exoc5 NEPHRIN Neph1 肾足细胞 功能紊乱 肾病综合征 

分 类 号：R363[医药卫生—病理学]