核因子κB受体活化因子通过TRPC6?NFATc1信号通路介导足细胞损伤  

Rank mediated podocyte injure by regulating TRPC6-NFATc1 axis

作　　者：张鸿 梁顺 杜玥 窦曹帅 张丽[2] 谈锦萍 刘双信[2] 梁馨苓[2] 章斌 ZHANG Hong;LIANG Shun;DU Yue;DOU Caoshuai;ZHANG Li;TAN Jinping;LIU Shuangxin;LIANG Xinling;ZHANG Bin(The Second School of Clinical Medicine,Southern Medical University,Guangzhou 510282,China)

机构地区：[1]南方医科大学第二临床医学院,广州510282 [2]广东省人民医院肾内科广东省医学科学院,广州510080

出　　处：《实用医学杂志》2019年第1期25-29,34,共6页The Journal of Practical Medicine

基　　金：国家自然科学基金面上项目(编号:81370808;81670656;81570642)

摘　　要：目的探究Rank通过TRPC6-NFATc1信号通路介导足细胞损伤。方法 (1)通过荧光定量PCR、Western印迹及免疫荧光染色检测Ionomycin刺激下足细胞Rank表达;(2)足细胞转染Rank-siRNA后,荧光定量PCR及Western印迹检测沉默效率,Western印迹检测足细胞标记蛋白Podocin表达;(3)通过Western印迹及免疫荧光染色检测沉默Podocin后足细胞核NFATc1表达;(4)Western印迹检测沉默Rank后TRPC6的表达,通过免疫共沉淀(Co-IP)检测Rank与TRPC6间关联。结果 (1)Ionomycin刺激下足细胞Rank表达增加;(2)沉默Rank表达后足细胞标记蛋白Podocin表达上调;(3)沉默Rank后足细胞核内NFATc1表达减少;(3)沉默Rank后足细胞TRPC6表达减少,Co-IP显示Rank结合TRPC6,且Ionomycin刺激下结合量表达增多。结论 Rank对足细胞具有损伤作用,其机制为Rank结合TRPC6调控NFATc1,从而介导足细胞损伤。Objective To explore the role of Rank in TRPC6-NFATc1 axis induced podocyte injury.Methods(1)After podocyte was exposed to ionomycin,real-time quantity PCR,western blotting and immunofluorescence staining were used to analyze the expression of Rank;(2)Real-time quantity PCR and western blotting were used to analyze the effect of Rank-siRNA,and western blotting was used to detect the expression of Podocin in Rank knockdown podocytes;(3)Western blotting and immunofluorescence staining were used to evaluate the change of nuclear localization of NFATc1 in Rank knockdown podocytes;(5)Co-immunoprecipitation assay was used to observe the relationship between Rank and TRPC6.Results(1)The mRNA and protein expression of Rank were markedly increased in injured podocyte after treatment with ionomycin;(2)Podocin protein expression was up-regulated in Rank knockdown podocytes;(3)After knockdown Rank,nuclear localization of NFATc1 was decreased;(4)Co-IP assay demonstrated that Rank binds to TRPC6 and ionomycin enhance the binding of Rank to TRPC6.Conclusion Rank mediates podocyte injure by binding to TRPC6,and enhances NFATc1signalling modulating podocyte injury.

关 键 词：足细胞 核因子ΚB受体活化因子 瞬时受体电位通道 活化T细胞核因子类 

分 类 号：R692[医药卫生—泌尿科学]