锌指蛋白281抑制高糖诱导的肾小管上皮细胞上皮间质转化和细胞外基质合成  

Zinc finger protein 281 inhibits high glucose-induced epithelial-mesenchymal transition and extracellular matrix synthesis in renal tubular epithelial cells

作　　者：侯维玲 乔云阳 吴小芸[2] 施会敏 曲高婷 张爱青[3] HOU Weiling;QIAO Yunyang;WU Xiaoyun;SHI Huimin;QU Gaoting;ZHANG Aiqing(Department of Pediatrics,the Affiliated Jiangning Hospital of Nanjing Medical University,Nanjing 211100,China;Department of Pediatrics,the Second Affiliated Hospital of Nanjing Medical University;Department of Pediatrics,the Fourth Affiliated Hospital of Nanjing Medical University)

机构地区：[1]南京医科大学附属江宁医院儿科,211100 [2]南京医科大学第二附属医院儿科 [3]南京医科大学第四附属医院儿科

出　　处：《天津医药》2024年第7期720-726,共7页Tianjin Medical Journal

基　　金：江苏省医学会儿科医学科研专项基金项目(SYH-32034-0073);南京市卫生科技发展专项资金项目(YKK23209)。

摘　　要：目的探讨锌指蛋白281(ZNF281)在高糖(HG)诱导的肾小管上皮细胞(RTECs)上皮间质转化(EMT)和细胞外基质(ECM)合成中的作用及机制。方法使用HG诱导RTECs以构建糖尿病肾病模型,分为Control组、HG组和Mannitol组,使用CCK-8检测细胞增殖活力。使用小干扰RNA(siRNA)在HG诱导的RTECs中敲低ZNF281的表达水平,分为Control组、HG组、HG+ZNF281 siRNA组和HG+ZNF281 vector组。使用腺苷单磷酸活化蛋白激酶(AMPK)激动剂阿卡地新(AICAR)活化AMPK,分为Control组、HG组、HG+AICAR组和HG+二甲基亚砜组。实时荧光定量PCR和蛋白免疫印迹法检测ZNF281、EMT和ECM合成相关指标表达水平。结果与Control组相比,HG组ZNF281、波形蛋白(vimentin)、α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)和Ⅰ型胶原蛋白(ColⅠ)的蛋白和mRNA表达水平升高,E-钙黏蛋白(E-cadherin)表达水平降低;与HG组相比,HG+ZNF281 siRNA组和HG+AICAR组的EMT和ECM合成相关指标的蛋白和mRNA表达水平发生明显变化,同时HG+AICAR组ZNF281的蛋白和mRNA表达水平较HG组降低;在使用AICAR和转染ZNF281过表达质粒共处理的细胞中,AICAR+ZNF281组vimentin、α-SMA、FN、ColⅠ的表达水平较空载组升高,E-cadherin表达水平降低。结论AMPK通过负向调控ZNF281的表达水平进而抑制HG诱导的RTECs中EMT和ECM合成。Objective To investigate the role and mechanism of zinc finger protein 281(ZNF281)in high glucose(HG)-induced epithelial-mesenchymal transition(EMT)and extracellular matrix(ECM)synthesis in renal tubular epithelial cells(RTECs).Methods HG induced RTECs were used to construct a diabetic kidney disease cell model,and cells were divided into the control group,the HG group and the mannitol group.Cell proliferation viability was detected by CCK-8.The expression of ZNF281 was knocked down in HG-treated RTECs using small interfering RNA(siRNA).HG-induced RTECs after knockdown of ZNF281 were divided into the control group,the HG group,the HG+ZNF281 siRNA group and the HG+ZNF281 vector group.Adenosine monophosphate-activated protein kinase(AMPK)was activated using AMPK agonist,acadexin(AICAR),and then cells were divided into the control group,the HG group,the HG+AICAR group and the HG+dimethyl sulfoxide group.The expression levels of ZNF281,EMT and ECM synthesis-related indexes were detected by qPCR and Western blot assay.Results Compared with the control group,the protein and mRNA expression levels of vimentin,α-smooth muscle actin(α-SMA),fibronectin(FN)and collagenⅠ(ColⅠ)were significantly higher,and the expression of E-cadherin was significantly lower in the HG group.Compared with the HG group,the protein and mRNA expression levels of EMT and ECM synthesis-related indexes were significantly changed in the HG+ZNF281 siRNA group and the HG+AICAR group.The protein and mRNA expression levels of ZNF281 were significantly reduced in the HG+AICAR group compared with the HG group.In cells co-treated with AICAR and transfected with ZNF281 plasmid,the expression levels of vimentin,α-SMA,FN and ColⅠwere significantly higher in the AICAR+ZNF281 group,and Ecadherin was significantly lower compared with that of the vector group.Conclusion AMPK inhibits EMT and ECM synthesis in HG-treated RTECs by negatively regulating the expression level of ZNF281.

关 键 词：糖尿病肾病 Tristetraprolin蛋白 上皮-间质转化 细胞外基质 AMP活化蛋白激酶类 肾小管间质纤维化 锌指蛋白281 

分 类 号：R587.24[医药卫生—内分泌] R34[医药卫生—内科学]