摘要
目的:观察高糖条件下Smad3对肾小管上皮细胞转分化的影响,探讨Smad3在高糖诱导肾脏纤维化中的作用。方法:将体外培养的肾小管上皮细胞分为正常糖组(NG组)、高糖组(HG组),Western blot鉴定细胞表型(E-cadherin及Desmin蛋白)及对高糖刺激后表型蛋白的变化;Western blot和Real-time PCR验证敲低Smad3质粒(Smad3-sh)、过表达Smad3质粒(OE-Smad3)转染肾小管上皮细胞细胞后Smad3蛋白及mRNA表达水平;分别在NG和HG条件下敲低或过表达肾小管上皮细胞Smad3蛋白,采用Western blot检测Smad3、p-Smad3、E-cadherin及Desmin蛋白表达水平。结果:NG组E-cadherin蛋白高表达,而Desmin表达较少;与NG组比较,HG刺激后E-cadherin蛋白表达显著减少(P<0.05),Desmin蛋白表达显著增加(P<0.05);敲低肾小管上皮细胞中Smad3后,E-cadherin蛋白表达显著增加(P<0.05),Desmin蛋白表达显著减少(P<0.05);过表达肾小管上皮细胞中Smad3后,E-cadherin蛋白表达显著减少(P<0.05),Desmin蛋白表达显著增加(P<0.05)。结论:Smad3能促进HG诱导肾小管上皮细胞转分化过程,从而促进高糖介导的纤维化发生。
Objective:To observe the effect of high glucose(HG)-mediated Smad3 expression on renal tubular epithelial cell transdifferentiation and explore the role of Smad3 expression in high glucose-induced renal fibrosis.Methods:Renal tubular epithelial cells were grew in media supplemented with normal glucose(NG group)and HG(HG group),respectively.Western blot was used to assess the protein expression levels.The mRNA expression levels of genes was assessed by real-time PCR.Smad3-shRNA(Smad3-sh)plasmid and Smad3-expressing plasmid were transfected to inhibit and overexpress Smad3 in renal tubular epithelial cells,respectively.Results:Relative to NG group,HG treatment remarkably downregulated E-cadherin expression(P<0.05),but upregulated Desmin(P<0.05).Knockdown of Smad3 increased E-cadherin expression level(P<0.05)and decreased Desmin expression level(P<0.05),while overexpressing Smad3 reversed Smad knockdown-mediated effect on E-cadeherin and Desmin.Conclusion:Smad3 can promote the HG-mediated transdifferentiation of renal tubular epithelial cells,which leads to fibrosis.
作者
毛彦稳
张小欢
刘玲伶
刘慧铭
梁露群
张会芳
向珈谊
王圆圆
MAO Yanwen;ZHANG Xiaohuan;LIU Lingling;LIU Huiming;LIANG Luqun;ZHANG Huifang;XIANG Jiayi;WANG Yuanyuan(Department of Pathophysiology,Guizhou Medical University,School of Basic Medical,Laboratory of Pathogenesis Research,Drug Prevention and Treatment of Major Diseases,Guizhou Medical University,Guiyang 550025,Guizhou,China)
出处
《贵州医科大学学报》
CAS
2019年第12期1395-1400,共6页
Journal of Guizhou Medical University
基金
国家自然科学基金(81760131)
关键词
糖尿病肾病
高糖
SMAD3
肾小管上皮细胞
上皮细胞转分化
肾脏纤维化
diabetic nephropathy
high glucose
Smad3
renal tubular epithelial cells
epithelial cell transdifferentiation
renal fibrosis
