LncRNA H2k2促进高糖培养的肾小球系膜细胞增殖的研究

该研究主要探讨lncRNA H2k2对高糖培养的肾小球系膜细胞增殖的影响,采用qRTPCR检测lncH2k2在正常及糖尿病肾病小鼠肾脏组织中的表达,以及高低糖培养的系膜细胞中的表达;FISH与qRT-PCR检测lncH2k2的亚细胞定位;qRT-PCR检测lncH2k2过表达质粒及siRNA的转染效率;EdU检测转染lncH2k2过表达质粒或siRNA后系膜细胞增殖的变化。结果表明,lncH2k2在糖尿病肾病小鼠肾脏组织及高糖培养的系膜细胞中的表达升高,且lncH2k2主要分布于系膜细胞的细胞质中。在低糖培养的系膜细胞中转染lncH2k2过表达质粒后,与低糖培养的系膜细胞相比,过表达lncH2k2的低糖培养的系膜细胞增殖能力显著提高,并且将qRT-PCR检测筛选出的一条lncH2k2 siRNA转染到高糖培养的系膜细胞内,与高糖培养的系膜细胞相比,敲低lncH2k2后系膜细胞增殖能力显著降低。研究结果揭示,lncRNA H2k2在糖尿病肾病小鼠肾脏组织及系膜细胞中表达显著,lncRNA H2k2促进了系膜细胞增殖,这些结果表明,lncRNA H2k2可能参与了糖尿病肾病的发生发展。

LncRNA H2k2 Promotes Proliferation of Mesangial Cells Cultured in High Glucose

The aim of this study was to investigate the effect of lncRNA H2k2 on proliferation of glomerular mesangial cells in high glucose culture.qRT-PCR was used to detect the expressions of lncH2k2 in kidney tissues of diabetic nephropathy mice and normal tissues,and mesangial cells in high and low glucose.The subcellular localization of lncH2k2 was detected by FISH and nuclear isolation qRT-PCR.qRT-PCR detected the efficiency of lncH2k2 overexpression plasmid and siRNA.The proliferation of mesangial cells transfected with lncH2k2 overexpressed plasmids or siRNA was detected by EdU.The results showed that the expression of lncH2k2 was increased in renal tissues of diabetic nephropathy mice and in mesangial cells cultured with high glucose,and lncH2k2 was mainly distributed in the cytoplasm of mesangial cells.The mesangial cells proliferation ability was increased significantly after transfected lncH2K2 over expression plasmid,compared with that in low glucose mock group.Additionally,the best efficiency of siRNA lncH2k2 was detected by qRT-PCR,and the mesangial cells proliferation ability was decreased in siH2K2 group compared with that in high glucose mock group.The results revealed that lncRNA H2k2 was significantly expressed in mice renal tissue of diabetic nephropathy and mesangial cells,and also promoted the proliferation of mesangial cells.These results suggested that lncRNA H2k2 might be involved in the occurrence and development of diabetic nephropathy.