RanBPM对肾小管钠-氯共同转运子的调控作用及对ERK1/2信号通路的影响  

The regulatory role of RanBPM in renal tubular sodium chloride cotransporter and its effect on ERK1/2 signaling pathway

作　　者：张益前[1] 庄芝芝 沈猛 庄捷秋[2] 蔡晖[1] ZHANG Yiqian;ZHUANG Zhizhi;SHEN Meng;ZHUANG Jieqiu;CAI Hui(Department of Nephrology,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China;Department of Pediatric Nephrology,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院肾内科,浙江温州325027 [2]温州医科大学附属第二医院育英儿童医院儿童肾内科,浙江温州325027

出　　处：《温州医科大学学报》2020年第7期553-556,562,共5页Journal of Wenzhou Medical University

基　　金：国家自然科学基金资助项目(81170709)。

摘　　要：目的:观察Ran结合蛋白M(RanBPM)对肾小管钠-氯共同转运子(NCC)蛋白表达及相关ERK1/2信号通路蛋白表达的影响。方法:免疫共沉淀检测RanBPM蛋白与WNK4蛋白是否存在直接相互作用;观察转染RanBPM质粒对EGF诱导细胞ERK1/2磷酸化的影响,通过转染质粒过表达RanBPM蛋白或siRNA抑制RanBPM基因,免疫蛋白印迹法检测细胞内源性NCC蛋白含量和ERK1/2磷酸化水平的变化。结果:免疫共沉淀实验证实WNK4和RanBPM之间存在直接的相互作用。RanBPM可以抑制EGF诱导的ERK1/2磷酸化。RanBPM过表达可使细胞ERK1/2磷酸化减少(1.000±0.074 vs. 0.275±0.041,P<0.01),而NCC蛋白表达增加(1.000±0.115 vs.1.470±0.105,P<0.01)。siRNA沉默RanBPM基因后,ERK1/2磷酸化增加(1.000±0.194 vs. 2.301±0.220,P<0.01),NCC蛋白表达下降(1.000±0.223 vs. 0.556±0.132,P<0.01)。转染RanBPM可阻止WNK4对ERK1/2信号通路的影响和对NCC蛋白表达的抑制作用。结论:RanBPM通过与WNK4相互作用,影响ERK1/2信号通路对NCC蛋白表达的调控作用。Objective:To observe the effect of Ran binding protein M(RanBPM)on the expression of NCC protein in renal tubules and the change of ERK1/2 signaling pathway protein.Methods:Immunocoprecipitation was used to detect whether there was direct interaction between RanBPM and WNK4 protein.The effect of transfection of RanBPM plasmid on ERK1/2 phosphorylation induced by EGF was observed.RanBPM gene was inhibited by overexpression of RanBPM protein or siRNA.The content of endogenous NCC protein and the level of ERK1/2 phosphorylation were detected by Western blot.The effect of RanBPM on WNK4 regulation of NCC by ERK1/2 phosphorylation was explored before and after transfection.Results:Immunocoprecipitation confirmed that there was a direct interaction between WNK4 and RanBPM.RanBPM could inhibit EGF induced ERK1/2 phosphorylation.Overexpression of RanBPM reduced ERK1/2 phosphorylation(1.000±0.074 vs.0.275±0.041,P<0.01)and increased NCC protein expression(1.000±0.115 vs.1.470±0.105,P<0.01).After siRNA silenced RanBPM gene,ERK1/2 phosphorylation increased(1.000±0.194 vs.2.301±0.220,P<0.01),resulting in decreased NCC protein expression(1.000±0.223 vs.0.556±0.132,P<0.01).When RanBPM was transfected,WNK4 inhibited NCC protein expression and no longer affected ERK1/2 phosphorylation.Conclusion:Ran BPM interacted with WNK4 affects the regulation of ERK1/2 signaling pathway in NCC protein expression.

关 键 词：Ran结合蛋白M WNK激酶 钠-氯共同转运子 ERK1/2信号传导通路 

分 类 号：R572.2[医药卫生—消化系统]