槲皮素通过ACE2-Ang-(1-7)-Mas轴对AngⅡ诱导下大鼠心肌收缩蛋白表达的影响
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篇名: 槲皮素通过ACE2-Ang-(1-7)-Mas轴对AngⅡ诱导下大鼠心肌收缩蛋白表达的影响
TITLE: Effects of Quercetin on the Expression of Ang Ⅱ-induced Myocardial Contractile Protein of Rats through ACE 2- Ang-(1-7)-Mas Axis
摘要: 目的:探讨槲皮素(Que)通过血管紧张素转换酶2-血管紧张素-(1-7)-Mas轴[ACE2-Ang-(1-7)-Mas轴]对血管紧张素Ⅱ(AngⅡ)诱导下原代大鼠心肌收缩蛋白表达的影响。方法:取1~2d日龄大鼠的心肌组织,分离、培养原代心肌细胞,并构建心肌细胞ACE2基因沉默模型。实验分为12组,其中AngⅡ组、AngⅡ+小干扰RNA(siRNA)组、AngⅡ+A779组为模型组,AngⅡ+氯沙坦组为阳性对照组,AngⅡ+Que40组、AngⅡ+Que80组、AngⅡ+siRNA+Que40组、AngⅡ+siRNA+Que80组、AngⅡ+A779+Que40组、AngⅡ+A779+Que80组为实验组,另设空白组、siRNA组。其中AngⅡ终浓度为1×10-6mol/L,siRNA终浓度为50nmol/L,Que终浓度分别为40、80μmol/L,A779(Mas受体拮抗剂)终浓度为1μmol/L,氯沙坦终浓度为1×10-4mol/L。分别检测各组心肌细胞中ACE2、Ang-(1-7)、Mas的mRNA和蛋白表达水平,以及Na+/Ca2+交换通道(NCX)、钙泵(SERCA2a)、磷蛋白(PLB)等3种心肌收缩蛋白的表达水平。结果:与AngⅡ组比较,AngⅡ+Que80组细胞中的MasmRNA表达水平升高(P<0.05);AngⅡ+氯沙坦组细胞中的ACE2、MasmRNA表达水平升高(P<0.05)。与AngⅡ组比较,AngⅡ+Que40组细胞中的ACE2、Ang-(1-7)蛋白表达水平升高(P<0.05);与AngⅡ+siRNA组比较,仅AngⅡ+siRNA+Que40组Ang-(1-7)蛋白表达水平升高(P<0.05);与AngⅡ+A779组比较,仅AngⅡ+A779+Que40组Ang-(1-7)蛋白表达水平升高(P<0.05)。与AngⅡ组比较,AngⅡ+Que40组NCX蛋白表达水平降低(P<0.05),AngⅡ+氯沙坦组NCX蛋白表达水平降低(P<0.05);与AngⅡ+A779组比较,AngⅡ+A779+Que80组NCX蛋白表达水平降低(P<0.05)。结论:Que一定程度上提高了AngⅡ诱导大鼠心肌细胞模型中ACE2-Ang-(1-7)-Mas轴的表达,从而调节心肌收缩蛋白。
ABSTRACT: OBJECTIVE:To in vestigate the effects of quercetin (Que)on the expressio n of angiotensin Ⅱ(AngⅡ)-induced myocardial contractile proteins of primary rats through angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas (ACE2-Ang- (1-7)-Mas)axis. METHODS :Cardiac tissue of rats aged 1-2 d were collected ,and primary cardiomyocytes were isolated and cultured. The gene silencing model of cardiomyocytes ACE2 was constructed. Experiments were divided into 12 groups. Among them,AngⅡ group,AngⅡ+ small interference RNA (siRNA)group,and Ang Ⅱ+ A 779 group were the model groups ;AngⅡ+ losartan group was positive control group ;AngⅡ+Que40 group,AngⅡ+Que80 group,AngⅡ+siRNA+Que40 group,AngⅡ+ siRNA+Que80 group,AngⅡ+A779+Que40 group and Ang Ⅱ+A779+Que80 group were the experimental groups ;blank group and siRNA group were set up. Ang Ⅱ concentration was 1×10-6 mol/L;siRNA final concentration was 50 nmol/L;Que concentration was 40 and 80 μmol/L;A779(Mas receptor inhibitor )concentration was 1 μmol/L;losartan concentration was 1×10-4 mol/L. mRNA and protein expression of ACE 2,Ang-(1-7) and Mas in primary cardiomyocytes were detected ;the expressions of myocardial contractile proteins were also determined ,such as Na +/Ca2+ exchange channel (NCX),calcium pump (SERCA2a), phosphoprotein (PLB). RESULTS :Compared with Ang Ⅱ group,mRNA expression of Mas was increased significantly in Ang Ⅱ + Que 80 group (P<0.05);mRNA expression of ACE2 and Mas were increased significantly in Ang Ⅱ + CZ0210-01) losartan group (P<0.05). Compared with Ang Ⅱ group,the 851136165@qq.com protein expression of ACE 2 and Ang- (1-7) were increased significantly in Ang Ⅱ+ Que 40 group(P<0.05);compared with Ang Ⅱ + siRNA group ,the protein expression of Ang-(1-7)were increased significantly in Ang Ⅱ+ siRNA+Que 40 group(P<0.05);compared with Ang Ⅱ+A779 group,the protein expression of Ang- (1-7)were increased significantly in Ang Ⅱ+A779+ Que 40 group(P<0.05). Compared with Ang Ⅱ group,the protein expression of NCX was decreased in Ang Ⅱ+Que40 group(P<0.05),protein expression of NCX was reduced in Ang Ⅱ+ losartan group (P<0.05);compared with Ang Ⅱ+A779 group,the protein expression of NCX was decreased in Ang Ⅱ+A779+ Que80 group (P<0.05). CONCLUSIONS :Que improves the expression of Ang Ⅱ -induced ACE 2-Ang-(1-7)-Mas axis in cardiomyocyte model to some extent ,so as to regulate myocardial contractile protein.
期刊: 2021年第32卷第23期
作者: 贾梦楠,朱明军,王永霞,李彬,郝轩轩,王新陆,于瑞,常鑫迪,李洁薇
AUTHORS: JIA Mengnan, ZHU Mingjun,WANG Yongxia,LI Bin,HAO Xuanxuan,WANG Xinlu,YU Rui,CHANG Xindi,LI Jiewei
关键字: 槲皮素;血管紧张素转换酶2-血管紧张素(1-7)-Mas轴;心肌细胞;血管紧张素Ⅱ;心肌收缩蛋白
KEYWORDS: Quercetin;ACE2-Ang-(1-7)-Mas axis ;Cardiomyocytes;Angiotensin Ⅱ;Myocardial contractile protein
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