丹蒌片对大鼠H9c2心肌细胞缺血再灌注损伤的保护作用及凋亡信号通路的影响  

Protective effect of Danlou Tablet against ischemia-reperfusion injury of H9C2 myocardial cells and apoptotic signaling pathway

作　　者：谭亚芳 祁建勇 梁玮婷 张敏州 TAN Ya-fang;QI Jian-yong;LIANG Wei-ting;ZHANG Min-zhou(2nd Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120,Guangdong,China)

机构地区：[1]广州中医药大学第二附属医院重症医学中医药应用研究团队,广东广州510120 [2]广州中医药大学中药学院,广东广州510006

出　　处：《广东医学》2019年第14期1983-1988,共6页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81473471);广东省中医院中医药科学技术研究专项(编号:YN2016QJ19)

摘　　要：目的通过构建H9c2心肌细胞缺氧模型,并基于线粒体通路和MAPK信号通路,探讨丹蒌片对心肌缺血损伤的保护作用。方法取对数生长期的大鼠H9c2心肌细胞,采用加入连二亚硫酸钠(Na2S2O4)的方法建立心肌细胞缺氧损伤模型,按空白对照组、模型组、抑制剂组和给药组,采用蛋白免疫印迹法分别检测心肌细胞凋亡蛋白Bcl-2、Bax、Caspase-3,心肌细胞相关蛋白ERK、p-ERK、JNK、p-JNK以及p38、p-p38的表达。结果与模型组相比,给药组能升高Bcl-2的表达,降低Bax、cleaved Caspase 3的表达,同时显著上调Bcl-2/Bax的比率(P<0.001);与抑制剂组相比,给药组能升高Bcl-2的表达,降低Bax、cleaved Caspase 3的表达,同时显著上调Bcl-2/Bax的比率(P<0.05)。与模型组相比,给药组能显著降低p-ERK、p-JNK和p-p38的蛋白水平(P<0.01);与抑制剂组相比,给药组能显著降低p-ERK和p-JNK的蛋白水平(P<0.01),而p-p38无明显差异(P>0.05)。结论丹蒌片对心肌缺血损伤的保护作用可通过调节线粒体通路抑制细胞凋亡,并与激活MAPK信号通路有关。Objective To establish the hypoxia model of H9c2 cardiomyocytes simulating myocardial ischemic injury at the cellular level;thus to investigate the effect of Danlou Tablet on sodium dithionite-induced H9c2 cardiomyocyte injury based on the mitochondrial pathway and MAPK signaling pathway. Methods In this study,a stable hypoxic injury model of cardiomyocytes was established by adding sodium hyposulfite ( Na2 S2O4 ) to simulate myocardial ischemic injury in vivo. Four groups of cell were studied,including Control group,I /R model group,inhibitor group and DLT group. Western Blot was applied for the assessment of myocardial apoptosis proteins Bcl-2,Bax,Caspase 3 and MAPK signaling pathway proteins,including ERK,p-ERK,JNK,p-JNK and p38,and p-p38. Results By adjusting the mitochondrial pathway,compared with the model group,the administration group could increase the expression of Bcl-2,reduce the expression of Bax and cleaved Caspase 3,and significantly increase the ratio of Bcl-2 /Bax ( P < 0. 001). Compared with the treatment group,the administration group increased the expression of Bcl-2,reduced the expression of Bax and cleaved Caspase 3,and significantly increased the ratio of Bcl-2 /Bax ( P < 0. 05). By adjusting the MAPK signaling pathway,compared with the model group,the administration group could significantly reduce the protein levels of p-ERK,p-JNK and p-p38 ( P < 0. 01). Compared with the inhibitor group,the administration group could significantly reduce the protein levels of p-ERK and p-JNK ( P < 0. 01),but there was no significant difference in p-p38 ( P > 0. 05). Conclusion The protective effect of Danlou Tablet on myocardial ischemic injury is via inhibiting apoptosis through regulating the mitochondrial pathway,and is related to activation of MAPK signaling pathway.

关 键 词：丹蒌片 心肌缺血 丝裂原蛋白活化激酶 

分 类 号：R542.2[医药卫生—心血管疾病] R364.12[医药卫生—内科学]