参麦注射液对缺氧心肌细胞凋亡的影响

目的： 观察参麦注射液对体外培养的心肌细胞缺氧后细胞凋亡的影响，并探讨其可能机制。方法： 分离乳鼠心肌细胞，进行离体培养。用95%N2与5%CO2混合气体造成缺氧环境，建立体外细胞缺氧模型，使细胞分别缺氧6 h和12 h。参麦注射液治疗组于缺氧处理前24 h至缺氧结束加入5 mL/L参麦注射液进行干预。首先通过Annexin V-FITC/PI双染法，流式细胞仪检测细胞凋亡情况。继而使用MTT法观察心肌细胞线粒体活性的改变，并应用激光共聚焦显微镜观察线粒体膜电位，荧光酶标仪检测胞浆内caspase3、7的活性。结果： 随着缺氧时间的延长凋亡细胞增多，参麦注射液能增强缺氧细胞的线粒体活性，维持线粒体膜电位，抑制caspase3、7的激活，从而减少凋亡细胞（P<0.01）。结论： 缺氧可以通过线粒体途径诱发心肌细胞的凋亡，参麦注射液能减少缺氧后细胞凋亡的发生，其机制与维持线粒体膜电位稳定，抑制caspase酶激活，即抑制凋亡的线粒体途径有关。

The effect of Shenmai injection on cardiac myocyte apoptosis after hypoxia

AIM: To observe the effect of Shenmai injection,a chinese medicine,on apoptosis of cardiac myocytes after hypoxia.METHODS: Cardiac myocytes were separated from neonate rat heart and cultivated in vitro.Hypoxia condition was induced by mixture of 95%N2 and 5%CO2.Cells were exposed to hypoxia for 6 h or 12 h and treated with Shenmai injection(5 mL/L) from 24 h before hypoxia until the end of hypoxia.First,apoptosis was detected with Annexin V-FITC and PI staining by flowcytometry.Then,the activity of cardiac myocyte mitochondria was observed by MTT method.Mitochondria membrane potential and the activity of caspase 3,7 were also measured by laser scan microscopy and multi-detection microplate reader,respectively.RESULTS: The apoptotic cells became more and more with prolonged hypoxia.Shenmai injection enhanced mitochondria activity,kept membrane potential,inhibited the activation of caspase3,7 and then decreased apoptotic cells(P<0.01).CONCLUSION: Hypoxia induces apoptosis in cardiac myocytes by mitochondria pathway.Shenmai injection can decrease cardiomyocyte apoptosis after hypoxia,the mechanism is related to mitochondria membrane potential stabilization and caspase inhibition.