【摘要】 目的通过观测大鼠股四头肌细胞线粒体形态学、氧化磷酸化水平和线粒体动力学-自噬相关蛋白表达,探讨脾气虚骨骼肌细胞线粒体损伤的可能机制。方法 16只雄性SD大鼠随机分为正常组和脾气虚证模型组(模型组),每组8只;取股四头肌组织,透射电镜观察线粒体形态学变化,ELISA法检测ATP和反应性氧簇(reactive oxygen species, ROS)水平,JC-1法检测线粒体膜电位(△Ψm),Western blot法检测介导线粒体动力学-自噬机制相关蛋白表达。结果与正常组比较,模型组线粒体变小且排列松散,ATP和△Ψm水平明显下降,ROS水平显著上升,动力学相关蛋白1、线粒体分裂蛋白1和线融素1表达均明显增加,但PINK1、Parkin和微管相关蛋白轻链3(microtubule-associated protein 1 light chain 3, LC3)-II表达及LC3-II/I比值均明显下降。结论脾气虚股四头肌线粒体损伤与线粒体动力学-自噬机制异常有关。更多还原

【Abstract】 Objective To explore the pathogenesis of muscular mitochondrial damage in spleen qi deficiency by detecting mitochondrial morphology, oxidative phosphorylation.China and key protein expressions related to mitochondrial dynamics-mitophagy. 16 male SD rats were randomly divided into the normal group and spleen Qi deficiency model group(model group), 8 in each group. Methods Based on quadriceps femoris, transmission electron microscopy was used to observe mitochondrial morphology in quadriceps femoris, ELISA was utilized to detect mitochondrial ATP and ROS levels, JC-1 method was employed to detect mitochondrial membrane potentials(△Ψm), and Western blot was chosen to check expressions of proteins mediating mitochondrial dynamics-mitophagy. Results Compared with those in the control group, mitochondrial mass was small and loosely connected, ATP level and △Ψm were significantly decreased, ROS level and expressions of dynamin-related protein 1, mitochondrial fission protein 1 and mitofusin 1 were increased markedly, however, PINK1, Parkin and microtubule-associated protein 1 light chain 3(LC3)-II expressions and the ratio of LC3-II/I were reduced, significantly, in the model group. Conclusion Abnormality of mitochondrial dynamics-mitophagy mechanism might contribute to the mitochondiral damage of skeletal muscles in spleen Qi deficiency.更多还原