浆细胞瘤异位基因1表达下调对高糖环境下肾小球系膜细胞增殖的影响及机制

发布时间：2018-03-04 01:16

  本文选题：长链非编码RNA　切入点：浆细胞瘤异位基因　出处：《山东医药》2017年14期 　论文类型：期刊论文

【摘要】：目的观察长链非编码RNA(lncRNA)中的浆细胞瘤异位基因1(PVT1)表达下调对高糖环境下肾小球系膜细胞增殖及细胞外基质(ECM)相关蛋白表达的影响,探讨PVT1在糖尿病肾病发病中的作用机制。方法培养人肾小球系膜细胞,分为正常对照组、高糖组、高糖+control siRNA组、高糖+PVT1 siRNA组。正常对照组在5.55 mmol/L的低糖DMEM完全培养基中培养,高糖组、高糖+control siRNA组、高糖+PVT1 siRNA组均在25mmol/L的高糖DMEM完全培养基中培养。高糖+control siRNA组、高糖+PVT1 siRNA组分别转染control siRNA、PVT1 siRNA。分别培养0、24、48、72 h后,采用CCK-8法测算各组细胞增殖率。采用流式细胞术检测各组细胞G1、S期比例。采用qRT-PCR法检测各组细胞PVT1 mRNA表达。采用Western blot法检测各组细胞ECM相关蛋白纤维连接蛋白(FN)、转化生长因子β_1(TGF-β_1)及Ⅰ型纤溶酶原激活物抑制因子(PAT-1)蛋白的表达。结果高糖组细胞各时间点细胞增殖率均高于正常对照组,高糖+PVT1 siRNA组各时间点细胞增殖率均低于高糖组(P均0.05)。与正常对照组比较,高糖组细胞G1期比例减少、S期比例增多;与高糖组比较,高糖+PVT1 siRNA组G1比例增加、S期比例减少(P均0.05)。高糖组、高糖+control siRNA组PVT1 mRNA相对表达量均高于正常对照组,高糖+PVT1 siRNA组PVT1 mRNA相对表达量低于高糖组(P均0.05)。高糖组、高糖+control siRNA组、高糖+PVT1 siRNA组FN、TGF-β_1、PAT-1蛋白表达均高于正常对照组,高糖+PVT1 siRNA组FN、TGF-β_1、PAT-1蛋白表达均低于高糖组(P均0.05)。结论在高糖环境下,肾小球系膜细胞中PVT1表达明显增加,下调PVT1表达可明显抑制肾小球系膜细胞的增殖、抑制ECM相关蛋白的表达。
[Abstract]:Objective to investigate the effect of down-regulation of plasmacytoma ectopic gene 1pPVT1 on the proliferation of glomerular Mesangial cells and the expression of ECM related proteins in high glucose environment. Methods Human glomerular Mesangial cells were cultured and divided into normal control group, high glucose group and high glucose control siRNA group. High glucose PVT1 siRNA group was cultured in 5. 55 mmol/L low glucose DMEM complete medium, high glucose group, high glucose control siRNA group, high glucose PVT1 siRNA group were all cultured in 25 mmol / L high glucose DMEM complete medium, high glucose control siRNA group, high glucose control siRNA group, high glucose control siRNA group, high glucose control siRNA group, high glucose control siRNA group, high glucose PVT1 siRNA group were all cultured in 25 mmol / L high glucose DMEM complete medium. High glucose PVT1 siRNA group was transfected with control siRNA-PVT1 siRNA. The proliferation rate of each group was measured by CCK-8 method, the proportion of G1S phase was detected by flow cytometry, the expression of PVT1 mRNA was detected by qRT-PCR assay, and the ECM associated protein fibronectin fibronectin was detected by Western blot assay. Expression of TGF- 尾 1) and plasminogen activator inhibitor type I (PAT-1) protein in TGF- 尾 1. Results the cell proliferation rate in high glucose group was higher than that in normal control group at all time points. The cell proliferation rate of high glucose PVT1 siRNA group was lower than that of high glucose group at each time point (P < 0.05). Compared with the normal control group, the G 1 phase ratio of high glucose group was lower than that of high glucose group, and the proportion of S phase was increased in high glucose group. The relative expression of PVT1 mRNA in the high glucose group and high glucose control siRNA group was higher than that in the normal control group, and the relative expression of PVT1 mRNA in the high glucose PVT1 siRNA group was lower than that in the high glucose PVT1 siRNA group, and the relative expression level of PVT1 mRNA in the high glucose PVT1 siRNA group was lower than that in the high glucose group. In high glucose control siRNA group and high glucose PVT1 siRNA group, the expression of FN- TGF- 尾 1 PAT-1 protein was higher than that in normal control group, and the expression of FN- TGF- 尾 1 PAT-1 protein in high glucose PVT1 siRNA group was lower than that in high glucose group (P 0.05). Conclusion the expression of PVT1 in glomerular Mesangial cells increased significantly in high glucose environment. Down-regulation of PVT1 expression significantly inhibited the proliferation of glomerular Mesangial cells and the expression of ECM related proteins.
【作者单位】： 广州医科大学附属第二医院;
【基金】：广东省省级科技计划资助项目(2014A020212324)
【分类号】：R692

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