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1,25-（OH）2D3通过lncRNA-uc.412抑制TGF-β1诱导的系膜细胞增殖的机制研究

1,25-（OH）2D3 attenuating mesangial cell proliferation induced by TGF-β1 through lncRNA-uc.412

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【作者】 安梦如； 黄婵； 王锦； 王银妃； 盛晓静； 张爱青； 甘卫华； 殷勤；

【Author】 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;

【通讯作者】 殷勤;

【机构】 南京医科大学第二附属医院儿科；

【摘要】 目的:研究1,25二羟基维生素D3[1,25-（OH）2D3]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）2D3、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）2D3+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）;（2）TGF-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）2D3组的ki67、PCNA的表达下调,p27的表达上调（P<0.01）。结论:1,25-（OH）2D3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）2D3阻滞细胞周期及下调lncRNA-uc.412的表达水平有关。

【Abstract】 Objective:To investigate the effect and mechanism of 1,25-（OH）2D3 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）2D3 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:（1）Compared with the TGF-β1 group, the expression of ki67,PCNA and lncRNA-uc.412 was down-regulated after intervention of 1,25-（OH）2D3+TGF-β1,whereas p27 expression was up-regulated（P<0.01）and the cell proliferation rate was decreased（P<0.01）;（2）The 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）;（3）Compared 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）2D3,yet p27 gene expression was up-regulated（P<0.01）.Conclusion:1,25-（OH）2D3 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）2D3 and down-regulation of lncRNA-uc.412 expression level.

【关键词】 1,25二羟基维生素D3； 长链非编码RNA uc.412； 转化生长因子β1； 系膜细胞； 增殖；
【Key words】 1,25-（OH）2D3； lncRNA-uc.412； TGF-β1； mesangial cells； proliferation；
【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)
  • 【文献出处】 皖南医学院学报 ,Journal of Wannan Medical College , 编辑部邮箱 ,2021年04期
  • 【分类号】R692
  • 【下载频次】92
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