艾司洛尔对缺氧复氧诱导的心肌细胞AMPK/GSK-3β通路及凋亡的影响  

Effects of esmolol on AMPK/GSK-3β pathway and apoptosis of cardiomyocytes induced by anoxia and reoxygenation

作　　者：赵鑫[1] 蔡丹霞[2] 王贺[3] ZHAO Xin;CAI Dan-xia;WANG He(Emergency Department of the First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471003,China;Department of Cardiology,the First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471003,China;Department of Cardiology,the First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南科技大学第一附属医院急诊科,河南洛阳471003 [2]河南科技大学第一附属医院心内科,河南洛阳471003 [3]河南中医药大学第一附属院河南中医药大学第一附属院心内科,河南郑州450000

出　　处：《现代药物与临床》2021年第2期213-218,共6页Drugs & Clinic

基　　金：河南省中医药科学研究专项课题(2019ZY2139)。

摘　　要：目的探讨艾司洛尔对缺氧复氧诱导的心肌细胞凋亡的影响,并初步研究其作用机制。方法体外培养大鼠心肌细胞(H9c2),并建立缺氧/复氧(I/R)损伤模型,随机分为对照组、H/R组和艾司洛尔低、中、高剂量(0.2、5.0、25.0μmol/L)组;噻唑蓝(MTT)法检测各组H9c2细胞存活率变化情况;膜联蛋白V-异硫氰酸荧光素/碘化丙啶(AnnexinV-FITC/PI)法检测各组H9c2细胞凋亡情况;蛋白印迹分析法检测各组H9c2细胞Bcl-2相关X蛋白(Bax)、半胱天冬氨酸酶-3(Caspase-3)、Bcl-2蛋白及腺苷单磷酸激活的蛋白激酶(AMPK)、糖原合酶激酶3β(GSK-3β)磷酸化水平;实时荧光定量PCR(q RT-PCR)法检测各组H9c2细胞AMPK、GSK-3βmRNA表达情况。结果与对照组相比,H/R组H9c2细胞存活率、Bcl-2蛋白表达水平、AMPK、GSK-3β蛋白磷酸化水平显著降低(P<0.05),细胞凋亡率、Bax、Caspase-3蛋白表达水平显著升高(P<0.05);与H/R组相比,艾司洛尔低、中、高剂量组H9c2细胞存活率、Bcl-2蛋白表达水平、AMPK、GSK-3β蛋白磷酸化水平依次升高(P<0.05),细胞凋亡率、H9c2细胞Bax、Caspase-3蛋白表达水平依次降低(P<0.05),均呈剂量依赖性。结论艾司洛尔能够抑制I/R诱导下心肌细胞的凋亡,可能与激活AMPK/GSK-3β磷酸化水平相关。Objective To study the effect of esmolol on cardiomyocyte apoptosis induced by hypoxia/reoxygenation and its mechanism. Methods The rat cardiomyocytes(H9c2) were cultured in vitro, and the model of hypoxia/reoxygenation(I/R) injury was established. H9c2 cells was divided into control group, H/R group, esmolol low dose group, esmolol medium dose group and esmolol high dose group(0.2、5.0、25.0 μmol/L). The survival rate of H9c2 cells was measured by methyl thiazolyl tetrazolium(MTT) method. The apoptosis of H9c2 cells was detected by Annexin V-FITC/PI method. The phosphorylation levels of Bcl-2 related X protein(Bax), Caspase-3, Bcl-2 protein, adenosine monophosphate-activated protein kinase(AMPK) and glycosylase kinase-3β(GSK-3β) in H9c2 cells were detected by Western blotting. The expression of AMPK and GSK-3β m RNAs in H9c2 cells were detected by real-time fluorescent quantitative PCR(qRT-PCR). Results Compared with the control group, the survival rate, Bcl-2 protein expression level, AMPK and GSK-3β protein phosphorylation levels of H9c2 cells in H/R group were significantly lower(P < 0.05), and the apoptosis rate, Bax and caspase-3 protein expression levels were significantly higher(P < 0.05). Compared with H/R group, the survival rate, Bcl-2 protein expression level, AMPK and GSK-3β protein phosphorylation levels of H9c2 cells in esmolol low, middle and high dose groups were increased in turn(P < 0.05), and the apoptosis rate, Bax and caspase-3 protein expression levels of H9c2 cells were decreased in turn(P < 0.05). Meanwhile, all of them were dose-dependent. Conclusion Esmolol can inhibit the apoptosis of cardiomyocytes induced by I/R, which may be related to the activation of AMPK/GSK-3β phosphorylation.

关 键 词：艾司洛尔 缺氧复氧 心肌细胞 凋亡 腺苷单磷酸激活的蛋白激酶 糖原合酶激酶3Β 

分 类 号：R966[医药卫生—药理学]