目的 探討艾司洛爾對(duì)缺氧復(fù)氧誘導(dǎo)的心肌細(xì)胞凋亡的影響，并初步研究其作用機(jī)制。方法 體外培養(yǎng)大鼠心肌細(xì)胞（H9c2），并建立缺氧/復(fù)氧（I/R）損傷模型，隨機(jī)分為對(duì)照組、H/R組和艾司洛爾低、中、高劑量（0.2、5.0、25.0 μmol/L）組；噻唑藍(lán)（MTT）法檢測(cè)各組H9c2細(xì)胞存活率變化情況；膜聯(lián)蛋白V-異硫氰酸熒光素/碘化丙啶（Annexin V-FITC/PI）法檢測(cè)各組H9c2細(xì)胞凋亡情況；蛋白印跡分析法檢測(cè)各組H9c2細(xì)胞Bcl-2相關(guān)X蛋白（Bax）、半胱天冬氨酸酶-3（Caspase-3）、Bcl-2蛋白及腺苷單磷酸激活的蛋白激酶（AMPK）、糖原合酶激酶3β（GSK-3β）磷酸化水平；實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測(cè)各組H9c2細(xì)胞AMPK、GSK-3β mRNA表達(dá)情況。結(jié)果 與對(duì)照組相比，H/R組H9c2細(xì)胞存活率、Bcl-2蛋白表達(dá)水平、AMPK、GSK-3β蛋白磷酸化水平顯著降低（P<0.05），細(xì)胞凋亡率、Bax、Caspase-3蛋白表達(dá)水平顯著升高（P<0.05）；與H/R組相比，艾司洛爾低、中、高劑量組H9c2細(xì)胞存活率、Bcl-2蛋白表達(dá)水平、AMPK、GSK-3β蛋白磷酸化水平依次升高（P<0.05），細(xì)胞凋亡率、H9c2細(xì)胞Bax、Caspase-3蛋白表達(dá)水平依次降低（P<0.05），均呈劑量依賴性。結(jié)論 艾司洛爾能夠抑制I/R誘導(dǎo)下心肌細(xì)胞的凋亡，可能與激活A(yù)MPK/GSK-3β磷酸化水平相關(guān)。

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 mRNAs 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.

R966

河南省中醫(yī)藥科學(xué)研究專項(xiàng)課題（2019ZY2139）