比索洛尔预处理通过调控ERK1/2途径抑制缺氧/复氧诱导的心肌细胞凋亡和纤维化  

Bisoprolol pretreatment inhibits hypoxia/reoxygenation induced cardiomyocyte apoptosis and fibrosis by regulating ERK1/2 pathway

作　　者：吴媛媛[1] 高元标 冼笃标 吴岳畅 赵映 WU Yuan-yuan;GAO Yuan-biao;XIAN Du-biao;WU Yue-chang;ZHAO Ying(Department of Cardiothoracic Surgery,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院心胸外科,海南海口570102

出　　处：《现代药物与临床》2022年第11期2430-2436,共7页Drugs & Clinic

基　　金：海南省卫生健康行业科研项目(20A200060)。

摘　　要：目的探讨比索洛尔预处理对缺氧/复氧诱导的心肌细胞凋亡和纤维化的影响,并分析分子机制。方法构建缺氧/复氧细胞模型,将H9c2细胞分为对照组、模型组、比索洛尔组、细胞外信号调节激酶(ERK1/2)通路激活剂(LM22B-10)组。采用酶联免疫吸附测定法(ELISA)检测细胞中乳酸脱氢酶(LDH)和肌酸激酶同工酶(CK-MB)水平,CCK-8法检测细胞活力,流式细胞术检测细胞凋亡水平,免疫印迹法(Western blotting)法检测凋亡蛋白、纤维化蛋白和ERK1/2信号通路蛋白表达。结果与模型组比较,比索洛尔组H9c2细胞中LDH和CK-MB水平、细胞凋亡率,切割型半胱氨酸天冬氨酸蛋白水解酶-3(cleaved Caspase-3)、B淋巴细胞瘤-2相关蛋白(Bax)、I型胶原(ColⅠ)、Ⅲ型胶原(ColⅢ)、α-平滑肌肌动蛋白(α-SMA)、基质金属蛋白酶9(MMP-9)蛋白表达以及磷酸化细胞外信号调节激酶1/2(p-ERK1/2)/(ERK1/2)比值显著降低;细胞活力,B淋巴细胞瘤2(Bcl2)、金属蛋白酶组织抑制因子1(TIMP-1)蛋白表达显著升高(P<0.05)。与比索洛尔组比较,LM22B-10组H9c2细胞中LDH和CK-MB水平、细胞凋亡率,Cleaved Caspase-3、Bax、ColⅠ、ColⅢ、α-SMA、MMP-9蛋白表达以及(p-ERK1/2)/(ERK1/2)比值均显著升高,细胞活力,Bcl-2、TIMP-1蛋白表达均显著降低(P<0.05)。结论比索洛尔预处理通过抑制ERK1/2信号通路活化减轻缺氧/复氧诱导的心肌细胞凋亡和纤维化。Objective To investigate the effects of bisoprolol pretreatment on cardiomyocyte apoptosis and fibrosis induced by hypoxia reoxygenation, and to analyze the its mechanisms. Methods Hypoxia/reoxygenation cell model were constructed, H9c2 cells were divided into control group, model group, bisoprolol group, and LM22B-10 group. LDH and CK-MB levels in cells were detected by enzyme-linked immunosorbent assay(ELISA), cell viability was detected by CCK-8 assay, and cell apoptosis was detected by flow cytometry. The expressions of apoptotic proteins, fibrotic proteins and ERK1/2 signaling pathway proteins were detected by Western blotting. Results Compared with model group, LDH and CK-MB levels and apoptosis rate of H9c2 cells, protein expression of cleaved Caspase-3, Bax, Col Ⅰ, Col Ⅲ, α-SMA, MMP-9, and(p-ERK1/2)/(ERK1/2) ratio were significantly decreased. Cell viability,Bcl-2, and TIMP-1 protein expressions were significantly increased in bisoprolol group(P < 0.05). Compared with bisoprolol group,LDH and CK-MB levels, apoptosis rate of H9c2 cells, protein expression of cleaved Caspase-3, Bax, Col Ⅰ, Col Ⅲ, α-SMA, MMP-9,and(p-ERK1/2)/(ERK1/2) ratio were significantly increased, and cell viability, Bcl-2 and TIMP-1 protein expression were significantly decreased in LM22B-10 group(P < 0.05). Conclusion Bisoprolol pretreatment attenuated hypoxia/reoxygenation induced cardiomyocyte apoptosis and fibrosis by inhibiting the activation of ERK1/2 signaling pathway.

关 键 词：比索洛尔 预处理 缺氧/复氧 心肌损伤 细胞凋亡 纤维化 细胞外信号调节激酶1/2 

分 类 号：R965[医药卫生—药理学]