Sos2抑制NFATc1介导的糖尿病肾病足细胞损伤的机制研究  

Mechanism of Sos2 inhibiting NFATc1-mediated podocyte injury in diabetic nephropathy

作　　者：杜玥 张鸿[2] 窦曹帅 谈锦萍 付蕾[2] 史伟[2] 梁馨苓[2] 刘双信[2] 章斌 DU Yue;ZHANG Hong;DOU Cao-shuai;TAN Jin-ping;FU Lei;SHI Wei;LIANG Xin-ling;LIU Shuang-xin;ZHANG Bin(School of Medicine,South China University of Technology,Guangzhou 510006,China;Department of Nephrology,Guangdong General Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China)

机构地区：[1]华南理工大学医学院,广东广州510006 [2]广东省人民医院肾内科,广东省医学科学院,广东广州510080

出　　处：《中国病理生理杂志》2018年第10期1834-1842,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81370808;No.81570642)

摘　　要：目的:观察高糖刺激对体外培养的小鼠肾足细胞鸟苷酸交换因子Sos2(Son of Sevenless homolog2)表达的影响,并初步探讨Sos2在高糖诱导足细胞损伤中的作用及其可能的分子机制。方法:通过免疫荧光染色及激光共聚焦显微镜观察Sos2在糖尿病肾病患者足细胞中的表达;体外培养小鼠永生化足细胞,以高糖(30 mmol/L葡萄糖)刺激足细胞48 h,采用RT-PCR、Western blot和免疫荧光检测高糖刺激下足细胞Sos2的mRNA及蛋白表达;采用Western bolt实验、免疫荧光及划痕实验检测Sos2过表达及沉默后podocin的表达、足细胞的活动性及NFATc1的入核情况;并采用RT-PCR检测NFATc1下游目的基因转录情况。结果:Sos2在糖尿病肾病患者的足细胞及高糖刺激体外培养的足细胞中表达显著降低(P <0. 05);沉默Sos2后,足细胞标志蛋白podocin表达显著降低,足细胞活动性增加,NFATc1入核增加,NFATc1下游目的基因转录增加(P <0. 05);与之相反,过表达Sos2组podocin表达显著增高,足细胞的活动性降低,NFATc1的入核减少,NFATc1下游目的基因转录降低(P <0. 05)。结论:Sos2可能通过抑制NFATc1入核而减轻糖尿病肾病足细胞损伤。AIM:To observe the effect of high glucose(HG)stimulation on the expression of guanine nucleotide exchange factor Sos2(Son of Sevenless homolog 2)in mouse podocytes,and to explore the role of Sos2 in HG-induced podocyte damage and its possible molecular mechanisms.METHODS:The expression of Sos2 in the podocytes of diabetic nephropathy patients was observed by immunofluorescence staining and laser confocal microscopy.In vitro,the Sos2 expression at mRNA and protein levels in immortalized podocytes with HG(30 mmol/L glucose)stimulation for 48 h was determined by the methods of RT-PCR,Western blot and immunofluorescence.Using Western blot,immunofluorescence and wound-healing assay,the expression of podocin,the translocation of NFATc1 into the nucleus and the podocyte migration with or without Sos2 silencing or overexpression were analyzed.The expression of downstream target genes for NFATc1 was detected by RT-PCR.RESULTS:The expression of Sos2 was significantly decreased in the podocytes of diabetic nephropathy patients and in vitro cultured podocytes with HG stimulation(P<0.05).When Sos2 was silenced,the expression of podocin was significantly decreased,the migration ability of podocytes was increased,and the translocation of NFATc1 into the nucleus was increased(P<0.05).In contrast,after overexpression of Sos2 in the podocytes with HG stimulation,the podocin expression level was obviously higher,and the podocyte migration ability and the translocation of NFATc1 into the nucleus were decreased(P<0.05).CONCLUSION:Sos2 may attenuate the diabetic nephropathy-induced podocyte injury by inhibiting NFATc1.

关 键 词：糖尿病肾病 足细胞 Sos2蛋白 NFATc1蛋白 

分 类 号：R587.2[医药卫生—内分泌] R363[医药卫生—内科学]