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目的:探讨参附注射液在抑制心肌细胞肥大过程中对线粒体内、外膜损伤的影响。方法:分离培养原代心肌细胞与血管紧张素Ⅱ(AngⅡ)共培养,BCA法检测细胞总蛋白含量、倒置显微镜拍摄并测量细胞直径,酶标仪检测线粒体外膜单胺氧化酶(MAO)活性、分光光度计检测线粒体内膜细胞色素C氧化酶(COX)活性和线粒体外膜损伤率、荧光显微镜测量线粒体内膜膜电位(ΔΨm)、SDS-聚丙烯酰胺凝胶电泳检测线粒体外膜Bcl-2、内膜ANT蛋白表达。在此基础上给予参附注射液,观察其对心肌细胞线粒体损伤的药理作用。结果:较同期模型组,100μl/ml浓度的参附注射液组能抑制心肌细胞肥大,降低MAO活性,升高COX活性,降低线粒体外膜损伤率,升高线粒体ΔΨm。但对于提高线粒体外膜Bcl-2蛋白和内膜ANT蛋白表达差异均无统计学意义。结论:参附注射液在抑制心肌细胞肥大的基础上,对线粒体内、外膜损伤有一定的改善作用。
Objective: To investigate the effect of Shenfu injection on the mitochondrial inner membrane and outer membrane in the process of inhibiting cardiomyocyte hypertrophy. Methods: Primary cultured cardiomyocytes were isolated and cultured with angiotensin Ⅱ (Ang Ⅱ). Total cellular protein (BCA) was assayed by BCA method. The diameter of the cells was measured by inverted microscope. The activity of monoamine oxidase (MAO) The activity of mitochondrial COX and the rate of mitochondrial outer membrane damage were measured. The mitochondrial inner membrane potential (ΔΨm) was measured by fluorescence microscopy. The mitochondrial outer membrane Bcl-2 was detected by SDS-polyacrylamide gel electrophoresis. Membrane ANT protein expression. On this basis, Shenfu injection was given to observe its pharmacological effects on mitochondrial damage of cardiomyocytes. Results: Compared with the model group, Shenfu injection group (100μl / ml) could inhibit cardiomyocyte hypertrophy, decrease MAO activity, increase COX activity, decrease mitochondrial outer membrane injury rate and increase mitochondrial ΔΨm. However, no significant difference was found between the expression of Bcl-2 protein and ANT protein in the endometrium. Conclusion: Shenfu injection inhibits cardiomyocyte hypertrophy based on the mitochondrial inner and outer membrane damage have some improvement.