摘要
目的:研究1,25二羟基维生素D 3[1,25-(OH)_(2)D_(3)]通过长链非编码RNA uc.412(lncRNA-uc.412)抑制转化生长因子β1(TGF-β1)诱导的大鼠系膜细胞增殖的可能机制。方法:用TGF-β1、1,25-(OH)_(2)D_(3)、SIS3分别干预系膜细胞,用lncRNA-uc.412过表达慢病毒转染系膜细胞,MTT法、流式细胞术检测系膜细胞增殖率,RT-PCR法检测ki67、p27、PCNA及lncRNA-uc.412的表达,Western blot检测smad3及p-smad3的表达。结果:①与TGF-β1组相比,1,25-(OH)_(2)D_(3)+TGF-β1干预后的ki67、PCNA、lncRNA-uc.412的表达下调,p27的表达上调(P<0.01),细胞增殖率下降(P<0.01);②TGF-β1+SIS3组p-smad3蛋白表达、lncRNA-uc.412的表达量较TGF-β1组下降(P<0.01);③与过表达lncRNA-uc.412组比较,过表达lncRNA-uc.412+1,25-(OH)_(2)D_(3)组的ki67、PCNA的表达下调,p27的表达上调(P<0.01)。结论:1,25-(OH)_(2)D_(3)可抑制TGF-β1诱导的大鼠系膜细胞增殖,其机制可能与1,25-(OH)_(2)D_(3)阻滞细胞周期及下调lncRNA-uc.412的表达水平有关。
Objective:To investigate the effect and mechanism of 1,25-(OH)_(2)D_(3) in the proliferation of rat mesangial cells induced by transforming growth factor β1(TGF-β1)through long non-coding RNA uc.412(lncRNA-uc.412).Methods:Mesangial cells were initially treated by TGF-β1,1,25-(OH)_(2)D_(3) and SIS3,respectively,and then transfected via lncRNA-uc.412 overexpression lentivirus.MTT and flow cytometry were used to measure the mesangial cell proliferation.RT-PCR and Western blot were performed to detect the expression of ki67,p27,PCNA and lncRNA-uc.412 as well as the expression of smad3 and p-smad3,respectively.Results:①Compared with the TGF-β1 group,the expression of ki67,PCNA and lncRNA-uc.412 was down-regulated after intervention of 1,25-(OH)_(2)D_(3)+TGF-β1,whereas p27 expression was up-regulated(P<0.01)and the cell proliferation rate was decreased(P<0.01);②The expression of p-smad3 protein and lncRNA-uc.412 was significantly lower in the TGF-β1+SIS3 group than in the TGF-β1 group(P<0.01);③Compared with the overexpressed lncRNA-uc.412 group,the gene expression of ki67 and PCNA was down-regulated after intervention with over-expressed lncRNA-uc.412+1,25-(OH)_(2)D_(3),yet p27 gene expression was up-regulated(P<0.01).Conclusion:1,25-(OH)_(2)D_(3) can inhibit the proliferation of mesangial cells induced by TGF-β1 in rats.The potential mechanism may be related to the cell cycle arrested by 1,25-(OH)_(2)D_(3) and down-regulation of lncRNA-uc.412 expression level.
作者
安梦如
黄婵
王锦
王银妃
盛晓静
张爱青
甘卫华
殷勤
AN Mengru;HUANG Chan;WANG Jin;WANG Yinfei;SHENG Xiaojing;ZHANG Aiqing;GAN Weihua;YIN Qin(Department of Pediatrics,The Second Affiliated Hospital of Nanjing Medical University,Nanjing 210003,China)
出处
《皖南医学院学报》
CAS
2021年第4期315-318,323,共5页
Journal of Wannan Medical College
基金
国家自然科学基金面上项目(81970664,81670650)
江苏省“六个一工程”拔尖人才科研项目(LGY2018071)
江苏省妇幼健康重点人才(FRC201737)
江苏省妇幼健康科研项目(F201508)。
作者简介
安梦如(1995-),女,2018级硕士研究生,(电话)15896260703,(电子信箱)835815089@qq.com;通信作者:殷勤,女,副教授,硕士生导师,(电子信箱)njyinqin@163.com。
