microRNA-31对高糖条件下人足细胞上皮-间充质转分化的影响  

Effects of microRNA-31 on high-glucose induced epithelial-mesenchymal transition of podocytes

作　　者：乐颖 蒋科威 薛萌 张秀珍 袁凤易 Le Ying;Jiang Kewei;Xue Meng;Zhang Xiuzhen;Yuan Fengyi(Department of Endocrinology and Metabolism,Shenzhen People′s Hospital,Second Affiliated Hospital of Jinan University,the First Affiliated Hospital of Southern University of Science and Technology,Shenzhen 518020,China;Department of Geriatrics,Shenzhen People′s Hospital,Second Affiliated Hospital of Jinan University,the First Affiliated Hospital of Southern University of Science and Technology,Shenzhen 518020,China)

机构地区：[1]深圳市人民医院、暨南大学第二临床医学院、南方科技大学第一附属医院内分泌科,518020 [2]深圳市人民医院、暨南大学第二临床医学院、南方科技大学第一附属医院老年病科,518020

出　　处：《国际内分泌代谢杂志》2022年第5期354-359,共6页International Journal of Endocrinology and Metabolism

基　　金：广东省医学科研基金项目(A2021302);国家自然科学青年项目(81900378)。

摘　　要：目的探讨microRNA-31(miR-31)在高糖诱导足细胞上皮-间充质转分化(EMT)中的机制。方法将体外培养的人肾小球足细胞按不同糖浓度分为低糖组(LG)、高渗组(HM)和高糖组(HG);按是否转染过表达miR-31(miR-31 mimics)分为miR-31过表达组(miR-31m组)、阴性对照组(miR-NC组)和脂质体组(Mock组);按照是否转染沉默低氧诱导因子-1抑制剂(FIH-1)及沉默miR-31(miR-31 inhibitor)分为FIH-1沉默组(si-FIH-1组)、miR-31沉默组(miR-31i组)、FIH-1沉默+miR-31沉默组(si-FIH-1+miR-31i组)及阴性对照组(NC组)。采用实时聚合酶链式反应(qRT-PCR)和Western blot检测各组细胞低氧诱导因子-1抑制剂(FIH-1)、转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)的mRNA和蛋白表达水平;采用双荧光素酶靶标实验验证miR-31与FIH-1基因的靶向关系。结果与LG和HM组比较,HG组miR-31表达水平显著升高(F=146.8,P<0.01),FIH-1蛋白表达水平明显下降(F=54.23,P<0.01),而TGF-β1及α-SMA蛋白表达水平均显著升高(F=360.6,P<0.01;F=193.7,P<0.01)。双荧光素酶报告基因实验分析结果显示,FIH-1是miR-31的靶基因。si-FIH-1与miR-31i共同转染时,可以恢复si-FIH-1或miR-31i单独转染导致的FIH-1、TGF-β1、α-SMA的mRNA及蛋白表达变化。结论miR-31靶向调控FIH-1促进足细胞EMT,抑制miR-31的表达可减轻高糖诱导的足细胞EMT。Objective To investigate the mechanism of microRNA-31(miR-31)in high-glucose induced epithelial-mesenchymal transition(EMT)of podocytes.Methods According to different sugar concentrations,human glomerular podocytes cultured in vitro were divided into low glucose group(LG),hyperosmolar group(HM)and high glucose group(HG);according to whether transfected miR-31 mimics,podocytes were divided into miR-31m group,negative miR-NC group and Mock group;according to whether transfected si-FIH-1 and miR-31i,podocytes were divided into si-FIH-1 group,miR-31i group,si-FIH-1+miR-31i group and NC group.Western blot was used to detect the protein expression levels of FIH-1,TGF-β1,andα-SMA in podocytes of each group;qRT-PCR was used to detect the mRNA expression levels of miR-31,FIH-1,TGF-β1,andα-SMA in podocytes of each group;double luciferase target experiment was used to verify the targeting relationship between miR-31 and FIH-1 gene.Results Compared with the LG and HM group,the expression of miR-31 in the HG group was significantly increased(F=146.8,P<0.01),and the expression of FIH-1 was significantly decreased(F=54.23,P<0.01),and the expression of TGF-β1 andα-SMA were increased significantly(F=360.6,P<0.01;F=193.7,P<0.01).The results of the dual luciferase reporter gene experiment showed that FIH-1 was the target gene of miR-31.When si-FIH-1 and miR-31i were co-transfected,the changes in the mRNA and protein expression of FIH-1,TGF-β1,andα-SMA caused by si-FIH-1 or miR-31i alone were restored.Conclusion miR-31 can promote the EMT of podocytes by targeting FIH-1.Inhibiting miR-31 expression can reduce the EMT induced by high glucose in podocytes.

关 键 词：miR-31 低氧诱导因子-1抑制剂 糖尿病肾脏病 足细胞 上皮-间充质转分化 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]