miR-30a抑制Ang2诱导的大鼠心肌细胞系H9c2自噬

miR-30a inhibits autophagy of rat cardiomyocyte line H9c2 induced by angiotensin 2

目的探索miR-30a对血管紧张素2(Ang2)诱导的大鼠心肌细胞自噬的影响。方法1)将H9c2细胞分为对照组(control)、Ang2处理组(Ang22×10-6 mol/L)、去甲肾上腺素阳性对照组(NE 5×10-6 mol/L)。处理细胞48 h,RT-qPCR检测miR-30a表达;Western blot检测LC3、Beclin-1自噬蛋白表达。2)将miR-30a mimics、miR-30a inhibitor、miR-30a NC通过Lipofectamine 2000转染至H9c2细胞中培养24 h,荧光显微镜检测转染率;转染成功后在Ang2刺激基础上设立Ang2+miR-30a mimics组、Ang2+miR-30a inhibitor组、Ang2+miR-NC组。检测miR-30a干预对Ang2诱导H9c2心肌细胞自噬的影响。结果1)与对照组相比,Ang2处理组、NE阳性对照组miR-30a mRNA表达显著下调(P<0.05);而LC3、Beclin-1蛋白表达显著升高(P<0.001);2)与Ang2处理组相比,Ang2+miR-30a mimics组LC3、Beclin1蛋白表达显著降低(P<0.001)。结论Ang2能下调miR-30a表达,诱导H9c2心肌细胞自噬;miR-30a mimics能够抑制Ang2诱导的H9c2心肌细胞自噬。

Objective To explore the effect of miR-30a on autophagy of rat cardiomyocytes induced by angiotensin 2(Ang2).Methods 1)H9c2 cells were treated with Ang22×10-6 mol/L and NE 5×10-6 mol/L respectively for 48 h,miR-30a expression was measured by RT-qPCR,LC3 and Beclin-1 protein expression were detected by Western blot.2)miR-30a mimics,miR-30a inhibitor and miR-30a NC were transfected into H9c2 cells by Lipofectamine2000 and incubated for 24 h,the transfection efficiency was detected by fluorescence microscope.RT-qPCR and Western blot were applied to detect the effect of miR-30a on autophagy of H9c2 induced by angiotensin 2.Results 1)Comparing with the control group,the expression of miR-30a mRNA in Ang2 treatment group and NE group was significantly decreased(P<0.05).Protein expression of LC3 and Beclin-1 in Ang2 group and NE group was significantly increased(P<0.001).2)Comparing with Ang2 treatment group,the expression of LC3 and Beclin-1 protein in Ang2+miR-30a mimics group significantly decreased(P<0.001).ConclusionsAng2 reduces the expression of miR-30a and induces H9c2 cardiomyocyte autophagy,miR-30a mimics can inhibit Ang2 induced H9c2 autophagy.