摘要
为探索高尿酸环境诱导下的大鼠肾细胞(NRK-52E)中锌α2糖蛋白(ZAG)与细胞外信号调节激酶1/2(ERK1/2)和p38形成反馈通路调控肾细胞上皮间质转化(EMT)的机制,将NRK-52E分为正常培养组(N组)和高尿酸培养组(H组)(20 mg/dL的尿酸刺激48 h),每组中的细胞都分别进行ZAG的过表达转染和敲低,观察细胞中ZAG的表达水平与ERK1/2和p38信号通路的交互作用。结果发现:相较于N组,高尿酸环境下的NRK-52E中EMT相关分子的mRNA及蛋白质的表达均上调(P<0.05);在高尿酸环境培养下,上调ZAG会减少NRK-52E在该通路中丝裂原活化蛋白激酶激酶(MAPKK)、ERK1/2、p38、活化转录因子2(ATF2)及蛋白激酶B(PKB或Akt)mRNA的表达量(P<0.01),而ZAG的下调则会增加(P<0.05)。在蛋白层面上,相较于未转染组,ZAG上调组的MAPKK、p38和Akt的表达量减少(P<0.05、P<0.01、P<0.001);ZAG下调组的ERK1/2、p38和Akt的表达量增加(P<0.001、P<0.05、P<0.01)。结果表明:ZAG的转染上调会减少NRK-52E中与EMT相关的标志物波形蛋白(vimentin)和层黏连蛋白(laminin)mRNA的表达量;调控ZAG在NRK-52E中的表达量可以在ERK1/2和p38信号通路中起到积极作用,进而抑制肾细胞EMT。该研究为临床治疗高尿酸血症肾病(HN)提供了试验依据。
The aim of this paper is to explore the mechanism of zinc-α2-glycoprotein(ZAG)forming a feedback pathway with extracellular signal-regulated kinase 1/2(ERK1/2)and p38 to regulate epithelial mesenchymal transition(EMT)in rat renal epithelial cells(NRK-52E)induced by high uric acid environment,we divided NRK-52E into normal control group and high uric acid-induced group which was stimulated by 20 mg/dL uric acid for 48 h,and the cells in each group were transfected with overexpression of ZAG and knocked down respectively,to observe the interactions between ZAG expression level and ERK1/2 and p38 signaling pathways in the cells.The results revealed that the mRNA and protein expression of EMT-related molecules were elevated in rat kidney cells under high uric acid environment compared with the normal culture group(P<0.05);up-regulation of ZAG decreased the expression of mitogen-activated protein kinase kinase(MAPKK),ERK1/2,p38,activated transcription factor-2(ATF2)and protein kinase B(PKB or Akt)mRNA in this pathway in rat kidney epithelial cells under high uric acid environment culture(P<0.01),while the downregulation of ZAG increased(P<0.05).At the protein level,the expression of MAPKK,p38 and Akt decreased in the ZAG up-regulated group compared to the untransfected group(P<0.05,P<0.01,P<0.001);and the expression of ERK1/2,p38 and Akt was elevated in the ZAG down-regulated group(P<0.001,P<0.05,P<0.01).The results suggest that transfection up-regulation of ZAG decreases the expression of laminin and vimentin mRNA,the marker genes related to EMT in NRK-52E;regulation of ZAG expression in NRK-52E can play a positive role in ERK1/2 and p38 signaling pathways,and thus inhibit renal cell EMT.This study provides an experimental basis for clinical treatment of hyperuricemic nephropathy(HN).
作者
陈柄源
杨洁
樊凯璇
陈秉朴
CHEN Bingyuan;YANG Jie;FAN Kaixuan;CHEN Bingpu(School of Basic Medical Sciences,Youjiang Medical University for Nationalities,Baise 533000,China)
出处
《激光生物学报》
CAS
2024年第2期176-184,共9页
Acta Laser Biology Sinica
基金
广西自然科学基金项目(2020GXNSFAA297151)
右江民族医学院高层次人才科研项目(RZ2100000455)
广西高校中青年教师科研基础能力提升项目(2021KY0554)。
关键词
肾纤维化
上皮细胞间充质转化
尿酸性肾病
高尿酸
锌α2糖蛋白
nephrofibrotic:epithelial mesenchymal transformation of urine
uric acid nephropathy
hyperuricemia
zinc-α2-glycoprotein
