miR-34a-5p通过调控MDM4对糖尿病性白内障晶状体上皮细胞间质转化的影响  

Effect of miR-34a-5p on epithelial-mesenchymal transition of lens epithelial cells in diabetic cataract by regulating mouse double microbody 4

作　　者：王心淼 王玉珏 郑丽莹 张敬轩 陈晔 WANG Xinmiao;WANG Yujue;ZHENG Liying;ZHANG Jingxuan;CHEN Ye(Ophthalmology Department of Inner Mongolia Autonomous Region Cancer Hospital,Hohhot 010010,Inner Mongolia,China;Inner Mongolia Medical University Ophthalmology 2020 Graduate Student,Hohhot 010110,Inner Mongolia,China;Endocrinology Department of Inner Mongolia People’s Hospital,Hohhot 010017,Inner Mongolia,China)

机构地区：[1]内蒙古自治区肿瘤医院眼科,内蒙古自治区呼和浩特市010010 [2]内蒙古医科大学,内蒙古自治区呼和浩特市010010 [3]内蒙古自治区人民医院内分泌科,内蒙古自治区呼和浩特市010017

出　　处：《眼科新进展》2023年第11期858-862,共5页Recent Advances in Ophthalmology

摘　　要：目的 探讨miR-34a-5p通过调控鼠双微体4(MDM4)对糖尿病性白内障晶状体上皮细胞(LEC)间质转化(EMT)的影响。方法 体外培养LEC,对LEC进行转染及双荧光素酶检测。实验分为对照组、高糖组、miR-NC组和miR-34a-5p抑制物组。采用MTT法和细胞划痕实验检测LEC细胞增殖和迁移水平,采用酶联免疫吸附试验检测LEC中丙二醛(MDA)、超氧化物歧化酶(SOD)水平,采用RT-qPCR检测LEC中miR-34a-5p、MDM4、转化生长因子-β2(TGF-β2)mRNA水平,采用Western blot检测LEC中MDM4、TGF-β2、β-catenin和E-cadherin蛋白表达水平。结果 miR-34a-5p对MDM4有潜在的结合位点。与对照组和miR-NC组相比,miR-34a-5p抑制物组LEC中MDM4-MUT荧光素酶活性均明显降低(均为P<0.05),对照组和miR-NC组LEC中MDM4-MUT荧光素酶活性差异无统计学意义(P>0.05);对照组、miR-NC组及miR-34a-5p抑制物组LEC中MDM4-WT荧光素酶活性差异无统计学意义(P>0.05)。与对照组相比,高糖组、miR-NC组和miR-34a-5p抑制物组LEC细胞增殖率、划痕愈合率、MDA、miR-34a-5p、TGF-β2 mRNA、TGF-β2蛋白和β-catenin蛋白均增加,SOD、MDM4 mRNA和蛋白、E-cadherin蛋白均降低(均为P<0.05);与高糖组和miR-NC组相比,miR-34a-5p抑制物组LEC细胞增殖率、划痕愈合率、MDA、miR-34a-5p、TGF-β2 mRNA和蛋白、β-catenin蛋白均降低,SOD、MDM4 mRNA和蛋白、E-cadherin蛋白均增加(均为P<0.05);高糖组与miR-NC组LEC各指标差异均无统计学意义(均为P>0.05)。结论 miR-34a-5p在糖尿病性白内障LEC细胞模型中表达上调,下调miR-34a-5p可以靶向促进MDM4的表达,抑制高糖诱导的LEC的EMT进程。Objective To explore the effect of miR-34a-5p on epithelial-mesenchymal transition(EMT)of lens epithelial cells(LECs)in diabetic cataract by regulating mouse double microbody 4(MDM4).Methods LECs were cultured in vitro and transfected for detection by the double luciferase assay.LECs were divided into the control group,high glucose(HG)group,miR-NC group,and miR-34a-5p inhibitor group,and treated for 24 hours.The proliferation and migration of LECs were detected by MTT and cell scratch assays.The levels of malondialdehyde(MDA)and superoxide dismutase(SOD)in LECs were measured by enzyme-linked immunosorbent assay,the mRNA levels of miR-34a-5p,MDM4 and transforming growth factor-beta 2(TGF-β2)in LECs were measured by real-time quantitative polymerase chain reaction,and the protein levels of MDM4,TGF-β2,β-catenin and E-cadherin in LECs were measured by Western blot.Results miR-34a-5p had a potential binding site for MDM4.Compared with the control and miR-NC groups,the luciferase activity of MDM4-MUT in the miR-34a-5p inhibitor group significantly decreased(both P<0.05).There was no significant difference in MDM4-MUT luciferase activity between the control and miR-NC groups(P>0.05).There were no significant differences in MDM4-WT luciferase activity among the control,miR-NC and miR-34a-5p inhibitor groups(all P>0.05).Compared with the control group,LEC proliferation rate,scratch healing rate,and levels of MDA,miR-34a-5p,TGF-β2 mRNA,TGF-β2 protein andβ-catenin protein increased in the HG,miR-NC and miR-34a-5p inhibitor groups,while the levels of SOD,MDM4 mRNA,MDM4 protein and E-cadherin protein decreased(all P<0.05).Compared with the HG and miR-NC groups,LEC proliferation rate,scratch healing rate,and the levels of MDA,miR-34a-5p,TGF-β2 mRNA,TGF-β2 protein andβ-catenin protein in the miR-34a-5p inhibitor group decreased,while the levels of SOD,MDM4 mRNA,MDM4 protein and E-cadherin protein increased(all P<0.05).There were no significant differences in said indexes between the HG and miR-NC groups(all P>0.05).Co

关 键 词：miR-34a-5p 鼠双微体4 糖尿病性白内障 间质转化 

分 类 号：R776.1[医药卫生—眼科]