miR-142-5p靶向调控TGF-β2促进人巨噬细胞凋亡

发布时间：2019-06-20 06:30

【摘要】：目的探讨miR-142-5p在动脉粥样硬化组织中的表达及对人巨噬细胞凋亡的作用。方法构建动脉粥样硬化大鼠模型,qRT-PCR检测动脉粥样硬化组织中miR-142-5p的表达水平。50μg/L氧化型低密度脂蛋白(ox-LDL)刺激人巨噬细胞、血管平滑肌细胞(VSMC)、内皮细胞24 h后,提取细胞RNA,qRT-PCR检测细胞中miR-142-5p的表达水平。靶基因预测软件预测miR-142-5p的靶基因,双荧光素酶报告基因鉴定靶基因的正确性。细胞转染miR-142-5p mimic、mimic control、miR-142-5p inhibitor、inhibitor control,Western blot和qRT-PCR检测miR-142-5p对靶基因的调控作用,流式细胞仪检测miR-142-5p和靶基因对巨噬细胞凋亡的作用。结果 miR-142-5p在动脉粥样硬化组织中表达上调,与正常组织相比差异显著(P0.01)。血管平滑肌细胞和内皮细胞经ox-LDL刺激后miR-142-5p的表达水平与刺激前没有明显变化,而巨噬细胞经ox-LDL刺激后miR-142-5p的表达水平较刺激前明显升高(P0.01)。预测miR-142-5p的靶基因为转化生长因子β2(TGF-β2)。miR-142-5p mimic与TGF-β2共转染后荧光素酶活性最低;miR-142-5p mimic组TGF-β2蛋白和mRNA的表达水平与mimic control组相比明显下降,miR-142-5p inhibitor组TGF-β2蛋白和mRNA的表达水平与inhibitor control组相比明显升高(P0.01);miR-142-5p mimic组细胞凋亡率明显高于mimic control组,TGF-β2 siRNA组细胞凋亡率明显高于siRNA control组(P0.01)。结论 miR-142-5p在动脉粥样硬化中过度表达,miR-142-5p通过下调靶基因TGF-β2促进人巨噬细胞凋亡。
[Abstract]:Objective to investigate the expression of miR-142-5p in atherosclerosis and its effect on apoptosis of human macrophages. Methods Atherosclerosis rat model was established. The expression of miR-142-5p in atherosclerotic tissue was detected by qRT-PCR. 50 渭 g / L oxidized low density lipoprotein (ox-LDL) stimulated human macrophages and vascular smooth muscle cell (VSMC), endothelial cells for 24 h. The expression of miR-142-5p in vascular smooth muscle cells was detected by RNA,qRT-PCR. The target gene prediction software predicted the correctness of target gene identification of miR-142-5p target gene and double luciferase reporter gene. The regulatory effect of miR-142-5p on target gene was detected by miR-142-5p mimic,mimic control,miR-142-5p inhibitor,inhibitor control,Western blot and qRT-PCR, and the effect of miR-142-5p and target gene on macrophage apoptosis was detected by flow cytometry. Results the expression of miR-142-5p in atherosclerotic tissues was up-regulated, which was significantly different from that in normal tissues (P 0.01). The expression level of miR-142-5p in vascular smooth muscle cells and endothelial cells stimulated by ox-LDL had no significant change compared with that before stimulation, but the expression level of miR-142-5p in macrophages stimulated by ox-LDL was significantly higher than that before stimulation (P 0.01). The target of miR-142-5p was predicted to be transforming growth factor 尾 2 (TGF- 尾 2). The luciferase activity of miR-142-5p mimic and TGF- 尾 2 co-transfected group was the lowest, the expression level of TGF- 尾 2 protein and mRNA in miR-142-5p mimic group was significantly lower than that in mimic control group, and the expression level of TGF- 尾 2 protein and mRNA in miR-142-5p inhibitor group was significantly higher than that in inhibitor control group (P 0.01). The apoptosis rate of miR-142-5p mimic group was significantly higher than that of mimic control group, and the apoptosis rate of TGF- 尾 2 siRNA group was significantly higher than that of siRNA control group (P 0.01). Conclusion miR-142-5p is overexpressed in atherosclerosis. MiR-142-5p promotes apoptosis of human macrophages by down-regulating the target gene TGF- 尾 2.
【作者单位】：郑州大学附属南阳中心医院特需病房一病区;
【分类号】：R543.5

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