TGF-β_(1)对大鼠肾小球系膜细胞纤维化蛋白及长链非编码RNA uc.412表达的影响  

Effects of TGF-β_(1) on rat glomerular mesangial cell fibrosis-related protein and lncRNA uc.412 expression

作　　者：杨思慧 王锦 黄婵 安梦如 张爱青[1] 甘卫华[1] YANG Sihui;WANG Jin;HUANG Chan;AN Mengru;ZHANG Aiqing;GAN Weihua(The Second Affiliated Hospital of Nanjing Medical University,Nanjing 210003,China)

机构地区：[1]南京医科大学第二附属医院,南京210003

出　　处：《山东医药》2021年第14期6-9,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81670650)。

摘　　要：目的观察转化生长因子β_(1)(TGF-β_(1))对大鼠肾小球系膜细胞纤维化蛋白的影响,并探讨其对长链非编码RNA uc.412(lncRNA uc.412)的调控作用。方法体外培养肾小球系膜细胞,将细胞随机分为3组,对照组仅加入培养基,TGF-β_(1)组加入TGF-β_(1),TGF-β_(1)+SIS3组同时加入TGF-β_(1)和TGF-β下游蛋白Smad3磷酸化特异性抑制剂SIS3,干预24 h。采用Real-time PCR法检测各组lncRNA uc.412表达;Western blotting法检测各组细胞纤维化相关蛋白1型胶原蛋白(Collagen-1)、α平滑肌肌动蛋白(α-SMA)、p-Smad3蛋白表达。利用慢病毒过表达lncRNA uc.412后转染肾小球系膜细胞细胞,建立lncRNA uc.412过表达组,并设病毒空载组和正常对照组。采用Real-Time PCR法检测Collagen-1、α-SMA mRNA表达,Western blotting法检测Collagen-1、α-SMA蛋白表达。结果与对照组相比,TGF-β_(1)组lncRNA uc.412表达升高(P<0.01),Collagen-1、α-SMA及p-Smad3蛋白表达升高(P均<0.05);与TGF-β_(1)组相比,TGF-β_(1)+SIS3组lncRNA uc.412表达下调(P<0.01),Collagen-1、α-SMA及p-Smad3蛋白表达降低(P均<0.05)。构建lncRNA uc.412过表达系膜细胞模型后,与对照组比较,过表达组α-SMA、Collagen-1蛋白及mRNA表达均升高(P均<0.05)。结论TGF-β_(1)通过下游Smad3信号通路上调lncRNA uc.412表达,诱导肾小球系膜细胞纤维化蛋白表达增加。Objective To observe the effect of transforming growth factor-β_(1)(TGF-β_(1))on rat glomerular mesangial cell fibrosis-related protein and to investigate its regulation on long non-coding RNA uc.412(lncRNA uc.412).Meth⁃ods The rat glomerular mesangial cells were cultured in vitro and then were divided into three groups and intervented for 24 h:cells in the control group were treated with culture medium,the TGF-β_(1) group with TGF-β_(1),and the TGF-β_(1)+SIS3 group with TGF-β_(1) and SIS3.Real-time PCR was used to detect the expression of lncRNA uc.412 in each group;Western blotting was used to detect the expression of fibrosis-related protein Collagen-1(Collagen-1),α-smooth muscle actin(α-SMA)and p-Smad3 protein in each group.We used lentivirus to overexpress lncRNA uc.412 and then transfected mesan⁃gial cells to establish lncRNA uc.412 overexpression group,and set up virus empty group and normal control group.Real-Time PCR was used to detect Collagen-1 andα-SMA mRNA expression,and Western blotting to detect Collagen-1 andα-SMA protein expression.Results Compared with the control group,the expression level of lncRNA uc.412 was signifi⁃cantly up-regulated,and p-Smad3,Collagen-1,andα-SMA protein levels increased in the TGF-β1 group(all P<0.05).Compared with the TGF-β_(1) group,the expression level of lncRNA uc.412 was significantly down-regulated,and the ex⁃pression levels of p-Smad3,Collagen-1,andα-SMA protein in the TGF-β_(1)+SIS3 group decreased(all P<0.05).Ln⁃cRNA uc.412 overexpressed mesangial cell model was successfully constructed.Compared with the control group,the ex⁃pression levels ofα-SMA and Collagen-1 protein and mRNA increased significantly in the overexpression group(all P<0.05).Conclusion TGF-β_(1) up-regulates the lncRNA uc.412 expression via Smad3/lncRNA uc.412 signaling pathway and thus induces the increase of protein expression related to mesangial cell fibrosis.

关 键 词：肾纤维化 肾小球系膜细胞 长链非编码RNA 转化生长因子β_(1) 

分 类 号：R692[医药卫生—泌尿科学]