miR-27a-3p靶向SnoN抑制高糖诱导的人近端肾小管上皮细胞EMT的作用  

miR-27a-3p inhibits EMT in high glucose-induced human proximal tubular epithelial cells by targeting SnoN

作　　者：彭君[1] 彭家清[1] 秦鹏[1] 罗先荣 张敏[1] PENG Jun;PENG Jiaqing;QIN Peng;LUO Xianrong;ZHANG Min(Department of Nephrology,Jingzhou Central Hospital,Jingzhou 433200,China)

机构地区：[1]荆州市中心医院肾内科

出　　处：《免疫学杂志》2020年第1期45-51,共7页Immunological Journal

基　　金：荆州市科技发展计项目（2015049）

摘　　要：目的研究miR-27a-3p对高糖诱导的人近端肾小管上皮细胞EMT的影响并探讨其机制。方法运用qRT-PCR检测HK2细胞中miR-27a-3p的表达;将HG+anti-miR-NC组(转染anti-miR-NC)、HG+anti-miR-27a-3p组(转染anti-miR-27a-3p)、HG+pcDNA组(转染pcDNA)、HG+pcDNA-SnoN组(转染pcDNA-SnoN),均用脂质体转染至HK2细胞,并用D-葡萄糖(30 mmol/L)处理48 h;Western blot检测各组细胞中E-cadherin、N-cadherin、SnoN、TGF-β1、p-Smad2的蛋白表达;双荧光素酶报告基因检测实验检测细胞的荧光活性。结果与低糖对照组相比,HG组细胞中miR-27a-3p显著升高,SnoN显著降低,E-cadherin、p-Smad2显著降低,N-cadherin、TGF-β1表达显著升高,(P<0.05);抑制miR-27a-3p、过表达SnoN均可上调E-cadherin,下调N-cadherin,抑制HK2细胞EMT;SnoN是miR-27a-3p的靶点。过表达SnoN可下调TGF-β1,上调p-Smad2,失活TGF-1信号通路。结论miR-27a-3p可抑制高糖诱导的人近端肾小管上皮细胞EMT,其机制可能与靶向SnoN失活TGF-β1信号通路有关,将可为糖尿病肾病的治疗提供依据。This study was designed to investigate the effect of miR-27a-3p on high glucose(HG)-induced EMT in human proximal tubular epithelial cells and to explore its mechanism.HK2 cells were treated with D-glucose(30 mmol/L)for 48 hours,and then transfected with anti-miR-NC(HG+anti-miR-NC group),anti-miR-27a-3p(HG+anti-miR-27a-3p group),pcDNA(HG+pcDNA group),or pcDNA-SnoN(HG+pcDNA-SnoN group)by using liposome.qRT-PCR was used to detect the expression of miR-27a-3p in HK2 cells;Western blot was used to detect the expression of E-cadherin,N-cadherin,SnoN,TGF-β1 and p-Smad2.The fluorescence activity of the cells was detected by double luciferase reporter gene assay.Compared with the low glucose control group,HG group of HK2 cells demonstrated higher levels of miR-27a-3p,N-cadherin,and TGF-β1,while lower levels of SnoN,E-cadherin,and p-Smad2(P<0.05).Both miR-27a-3p inhibition and SnoN overexpression could up-regulate E-cadherin,down-regulate N-cadherin,and inhibit HK2 cell EMT.SnoN is a target of miR-27a-3p.The overexpression of SnoN down-regulated TGF-β1,up-regulated p-Smad2,and inactivated TGF-β1 signaling pathway.In conclusion,miR-27a-3p can inhibit high glucose-induced EMT in human proximal tubular epithelial cells.The mechanism may be related to TGF-β1 signaling pathway inactivation by targeting SnoN,which may provide a basis for the treatment of diabetic nephropathy.

关 键 词：miR-27a-3p SNON EMT TGF-β1信号通路 

分 类 号：R334+.1[医药卫生—人体生理学]