miR-30a抑制Ang2诱导的大鼠心肌细胞系H9c2自噬

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (4): 490-494.

miR-30a抑制Ang2诱导的大鼠心肌细胞系H9c2自噬

于甜乐¹, 马晗², 吴新华^3,4,5, 陈章荣^3,4,5, 孙彪¹, 彭媛¹, 刘宏^3,4,5*

1.大理大学临床医学院, 云南大理 671000;
2.云南省滇东北区域医疗中心呼吸内科, 云南昭通 657000;
3.大理大学第一附属医院心血管内科, 云南大理 671000;
4.云南省跨高原心血管疾病防治工程研究中心,云南大理 671000;
5.大理大学跨高原心血管疾病防治研究所,云南大理 671000

收稿日期:2020-09-18 修回日期:2020-12-31 出版日期:2021-04-05 发布日期:2021-04-05
通讯作者: ^*daliliuhong@163.com
基金资助:
国家自然科学基金(81760085)

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

YU Tian-le¹, MA Han², WU Xin-hua^3,4,5, CHEN Zhang-rong^3,4,5, SUN Biao¹, PENG Yuan¹, LIU Hong^3,4,5*

1. Clinical Medical College, Dali University, Dali 671000;
2. Department of Respiratory Medicine, Northeast Yunnan Regional Medical Center, Zhaotong 657000;
3. Department of Cardiology, the First Affiliated Hospital,Dali University, Dali 671000;
4. Yunnan Research Center for Prevention and Treatment of Trans-plateau Cardiovascular Disease, Dali 671000;
5. Institute of Trans-plateau Cardiovascular Disease Prevention and Treatment, Dali University, Dali 671000, China

Received:2020-09-18 Revised:2020-12-31 Online:2021-04-05 Published:2021-04-05
Contact: ^*daliliuhong@163.com

摘要/Abstract

摘要： 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(Ang2 2×10^－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。

关键词: 自噬, 血管紧张素2(Ang2), miR-30a

Abstract: 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 Ang2 2×10^－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). Conclusions Ang2 reduces the expression of miR-30a and induces H9c2 cardiomyocyte autophagy, miR-30a mimics can inhibit Ang2 induced H9c2 autophagy.

Key words: autophagy, angiotensin 2 (Ang2), miR-30a

中图分类号:

于甜乐, 马晗, 吴新华, 陈章荣, 孙彪, 彭媛, 刘宏. miR-30a抑制Ang2诱导的大鼠心肌细胞系H9c2自噬[J]. 基础医学与临床, 2021, 41(4): 490-494.

YU Tian-le, MA Han, WU Xin-hua, CHEN Zhang-rong, SUN Biao, PENG Yuan, LIU Hong. miR-30a inhibits autophagy of rat cardiomyocyte line H9c2 induced by angiotensin 2[J]. Basic & Clinical Medicine, 2021, 41(4): 490-494.

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