首页|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

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

扫码查看
原文链接
  • NETL
  • NSTL
  • 万方数据
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.
Zinc finger protein 281 inhibits high glucose-induced epithelial-mesenchymal transition and extracellular matrix synthesis in renal tubular epithelial cells
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 collagen Ⅰ(Col Ⅰ)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 Col Ⅰ were significantly higher in the AICAR+ZNF281 group,and E-cadherin 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.

diabetic nephropathiestristetraprolinepithelial-mesenchymal transitionextracellular matrixAMP-activated protein kinasestubulointerstitial fibrosiszinc finger protein 281

侯维玲、乔云阳、吴小芸、施会敏、曲高婷、张爱青

展开 >

南京医科大学附属江宁医院儿科(邮编 211100)

南京医科大学第二附属医院儿科

南京医科大学第四附属医院儿科

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

江苏省医学会儿科医学科研专项基金项目南京市卫生科技发展专项资金项目

SYH-32034-0073YKK23209

2024

10.11958/20240031

天津医药
天津市医学科学技术信息研究所

天津医药

CSTPCD
影响因子：1.107
ISSN：0253-9896
年,卷(期)：2024.52(7)
  • 1