上调微小RNA-142-3p对心肌肥厚中线粒体功能的影响
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摘要
目的观察微小RNA(microRNA,miR)-142-3p对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导的心肌肥厚中线粒体功能的影响。方法我们使用Sprague-Dawley(SD)大鼠的乳鼠心肌细胞,细胞培养后分成4组:空白组;AngⅡ组;miR nc+AngⅡ组;miR-142-3p mimic+AngⅡ组。分别往细胞中转染相同浓度的miR-142-3p和miR nc质粒6 h,实时定量聚合酶链反应(real-time polymerase chain reaction,rt-PCR)检测实验组miR-142-3p mRNA表达增多,提示细胞质粒转染成功,再用10-6mol/L浓度的AngⅡ诱导细胞48 h,使用线粒体MitoRed Tracker处理细胞30 min,共聚焦显微镜观察细胞中线粒体密度的变化;使用流式细胞仪检测线粒体膜电位变化。结果与空白组相比,AngⅡ组的线粒体膜电位减低(n=3,P<0.01);与AngⅡ+miR nc组相比,AngⅡ+miR-142-3p组的线粒体膜电位增加(n=3,P<0.01)。与空白组相比,AngⅡ组的线粒体荧光数量减低(n=3,P<0.01);与AngⅡ+miR nc组相比,AngⅡ+miR-142-3p组的线粒体荧光数量增加(n=3,P<0.01)。结论在AngⅡ诱导心肌肥大过程中miR-142-3p对心肌线粒体具有保护作用。
ObjectivesTo observe the effect of microRNA(miR)-142-3 p on mitochondrial function in angiotensin Ⅱ(Ang Ⅱ)-induced cardiac hypertrophy.MethodsWe used Sprague-Dawley(SD)neonatal rat cardiomyocytes. After cell culture,we divided them into 4 groups:control group;AngⅡ group;miR nc+AngⅡ group;miR-142-3 p mimic+AngⅡ group. The same concentration of miR-142-3 p and miR nc plasmids were transfected into the cells for 6 hours,and the expression of miR-142-3 p mRNA in experimental group was increased by real-time quantitative polymerase chain reaction(rt-PCR). It can be thought that the cell plasmid was successfully transfected. Then the cells were induced with AngⅡ at a concentration of 10-6 mol/L for 48 hours. The cells were treated with mitochondrial Mito-Red Tracker for 30 minutes. The mitochondrial density was observed by confocal microscopy. The mitochondrial membrane potential was detected by flow cytometry.ResultsCompared with control group,the mitochondrial membrane potential of the AngⅡgroup was decreased(n=3,P<0.01);compared with AngⅡ + miR nc induced group,the mitochondrial membrane potential was increased in the Ang Ⅱ + miR-142-3 p induced group(n=3,P<0.01). The number of mitochondrial fluorescence staining in Ang Ⅱ group was reduced compared with control group(n=3,P<0.01);compared with AngⅡ + miR nc induced group,the number of mitochondrial fluorescence staining was increased in AngⅡ + miR-142-3 p induced group(n=3,P<0.01).ConclusionsmiR-142-3 p has a protective effect on myocardial mitochondrial function during Ang Ⅱ-induce cardiac hypertrophy.
ObjectivesTo observe the effect of microRNA(miR)-142-3 p on mitochondrial function in angiotensin Ⅱ(Ang Ⅱ)-induced cardiac hypertrophy.MethodsWe used Sprague-Dawley(SD)neonatal rat cardiomyocytes. After cell culture,we divided them into 4 groups:control group;AngⅡ group;miR nc+AngⅡ group;miR-142-3 p mimic+AngⅡ group. The same concentration of miR-142-3 p and miR nc plasmids were transfected into the cells for 6 hours,and the expression of miR-142-3 p mRNA in experimental group was increased by real-time quantitative polymerase chain reaction(rt-PCR). It can be thought that the cell plasmid was successfully transfected. Then the cells were induced with AngⅡ at a concentration of 10-6 mol/L for 48 hours. The cells were treated with mitochondrial Mito-Red Tracker for 30 minutes. The mitochondrial density was observed by confocal microscopy. The mitochondrial membrane potential was detected by flow cytometry.ResultsCompared with control group,the mitochondrial membrane potential of the AngⅡgroup was decreased(n=3,P<0.01);compared with AngⅡ + miR nc induced group,the mitochondrial membrane potential was increased in the Ang Ⅱ + miR-142-3 p induced group(n=3,P<0.01). The number of mitochondrial fluorescence staining in Ang Ⅱ group was reduced compared with control group(n=3,P<0.01);compared with AngⅡ + miR nc induced group,the number of mitochondrial fluorescence staining was increased in AngⅡ + miR-142-3 p induced group(n=3,P<0.01).ConclusionsmiR-142-3 p has a protective effect on myocardial mitochondrial function during Ang Ⅱ-induce cardiac hypertrophy.
引文
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[11]WANG Y,OUYANG M,WANG Q,et al.MicroRNA-142-3p inhibits hypoxia/reoxygenationinduced apoptosis and fibrosis of cardiomyocytes by targeting high mobility group box 1[J].Int JMol Med,2016,38(5):1377-1386.2018-06-28
[2]DAI D F,JOHNSON S C,VILLARIN J J,et al.Mitochondrial oxidative stress mediates angiotensin II-induced cardiac hypertrophy and Galphaq overexpression-induced heart failure[J].Circ Res,2011,108(7):837-846.
[3]HUGEL S,HORN M,REMKES H,et al.Preservation of cardiac function and energy reserve by the angiotensinconverting enzyme inhibitor quinapril during postmyocardial infarction remodeling in the rat[J].J Cardiovasc Magn Reson.2001,3(3):215-225.
[4]ROSCA M G,TANDLER B,HOPPEL C L.Mitochondria in cardiac hypertrophy and heart failure[J].J Molec Cell Cardiol,2013,55:31-41.
[5]SUN Y L,LI S H,YANG L,et al.miR-376b-3p attenuates mitochondrial fission and cardiac hypertrophy by targeting MFF[J].Clin Exp Pharmacol Physiol,2018,45(8):779-787.
[6]ZHOU L Y,LIU J P,WANG K,et al.Mitochondrial function in cardiac hypertrophy[J].Int J Cardiol,2013,167(4):1118-1125.
[7]LU Y,WU F.A new miRNA regulator,miR-672,reduces cardiac hypertrophy by inhibiting JUN expression[J].Gene.2018,648:21-30.
[8]LOCK M C,TELLAM R L,BOTTING K J,et al.The role of miRNA regulation in fetal cardiomyocytes,cardiac maturation and the risk of heart disease in adults[J].J Physiol,2018,596(23):5625-5640.
[9]CHEN Z Y,CHEN F,CAO N,et al.miR-142-3p contributes to early cardiac fate decision of embryonic stem cells[J].Stem Cells Int,2017:1769298.
[10]SHARMA S,LIU J,WEI J,et al.Abstract 16798:MicroRNA142-5p targets myocardial hypertrophy and cytokine response[J].Circulation,2011,124:A16798.
[11]WANG Y,OUYANG M,WANG Q,et al.MicroRNA-142-3p inhibits hypoxia/reoxygenationinduced apoptosis and fibrosis of cardiomyocytes by targeting high mobility group box 1[J].Int JMol Med,2016,38(5):1377-1386.2018-06-28