Cheng Jie, Cheng Li, Tang Yubin, Li Huihua, Peng Wenfang, Huang Shan
Department of Endocrinology, Tongren Hospital, School of Medicine, Shanghai Jiao Tong University Shanghai, China.
Department of Gynecology, The Second Affiliated Hospital of Guiyang College of Traditional Chinese Medicine Guiyang, China.
Int J Clin Exp Pathol. 2018 Jul 1;11(7):3302-3309. eCollection 2018.
This study aimed to investigate the effect of the long non-coding RNA (lncRNA) Dlx6os1 inhibitor on cell proliferation, apoptosis and fibrosis in mouse mesangial cells (MMCs) under high glucose (HG) conditions. SV40 MES13 cells were cultured under 30 mmol/L glucose (HG group), 5.6 mmol/L glucose (normal glucose group, NG group) and 5.6 mmol/L glucose plus 24.4 mmol/L 3-O-methyl-D-glucose (osmotic control group, OC group), and expressions of lncRNA Dlx6os1, Gm13730, Rdh9, Chrm2 and Bex1 were determined by qPCR. NC-inhibitor plasmids and lncRNA Dlx6os1 inhibitor plasmids were transfected into SV40 MES13 cells cultured under HG conditions, and cell proliferation (at 0 h, 24 h, 48 h and 72 h), and the apoptosis rate (at 72 h), proteins and mRNAs expressions for proliferation and fibrosis markers (at 72 h) were detected by CCK-8, AV-PI, Western blot and qPCR assays, respectively. LncRNA Dlx6os1 was increased in the HG group compared with the OC and NG groups, while no difference of lncRNA Gm13730, Rdh9, Chrm2 or Bex1 was discovered among the three groups. The lncRNA Dlx6os1 inhibitor decreased cell proliferation at 24 h, 48 h and 72 h post plasmids transfection by the CCK-8 assay and reduced the expressions of Cyclin D1 and proliferating cell nuclear antigen (PCNA) mRNA and protein expressions compared with the NC-inhibitor. The cell apoptosis rate at 72 h increased compared with the NC-inhibitor. In addition, protein and mRNA expressions of markers for cell fibrosis (fibronectin and collagen I) were all decreased in the lncRNA Dlx6os1-inhibitor group compared to the NC-inhibitor group. In conclusion, inhibition of lncRNA Dlx6os1 decreases cell proliferation and fibrosis and increases cell apoptosis in diabetic nephropathy.
本研究旨在探讨长链非编码RNA(lncRNA)Dlx6os1抑制剂对高糖(HG)条件下小鼠系膜细胞(MMC)增殖、凋亡和纤维化的影响。将SV40 MES13细胞分别培养于30 mmol/L葡萄糖(HG组)、5.6 mmol/L葡萄糖(正常葡萄糖组,NG组)和5.6 mmol/L葡萄糖加24.4 mmol/L 3 - O - 甲基 - D - 葡萄糖(渗透对照组,OC组),通过qPCR检测lncRNA Dlx6os1、Gm13730、Rdh9、Chrm2和Bex1的表达。将NC - 抑制剂质粒和lncRNA Dlx6os1抑制剂质粒转染至HG条件下培养的SV40 MES13细胞,分别通过CCK - 8、AV - PI、Western blot和qPCR检测细胞增殖(0 h、24 h、48 h和72 h)、凋亡率(72 h)以及增殖和纤维化标志物的蛋白和mRNA表达(72 h)。与OC组和NG组相比,HG组lncRNA Dlx6os1表达增加,而lncRNA Gm13730、Rdh9、Chrm2或Bex1在三组间无差异。通过CCK - 8检测,lncRNA Dlx6os1抑制剂在质粒转染后24 h、48 h和72 h降低细胞增殖,与NC - 抑制剂相比,降低细胞周期蛋白D1和增殖细胞核抗原(PCNA)的mRNA和蛋白表达。与NC - 抑制剂相比,72 h时细胞凋亡率增加。此外,与NC - 抑制剂组相比,lncRNA Dlx6os1抑制剂组细胞纤维化标志物(纤连蛋白和I型胶原)的蛋白和mRNA表达均降低。总之,抑制lncRNA Dlx6os1可降低糖尿病肾病中的细胞增殖和纤维化,并增加细胞凋亡。