脾气虚大鼠股四头肌细胞线粒体动力学和自噬相关蛋白表达的研究  

Study of mitochondrial dynamics-mitophagy mechanism of rat quadriceps femoris in spleen Qi deficiency

作　　者：宋飞飞 范英丽[1] 刘慧慧[2] 刘旭东[2] 于化新[2] 单德红[2] SONG Fei-fei;FAN Ying-li;LIU Hui-hui;LIU Xu-dong;YU Hua-xin;SHAN De-hong(Chinese Medicine College of Hainan Medical University,Haikou 571199,China;Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]海南医学院中医学院,海南海口571199 [2]辽宁中医药大学,辽宁沈阳110847

出　　处：《时珍国医国药》2021年第8期2046-2048,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(81673851)。

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。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.

关 键 词：脾气虚 骨骼肌 线粒体 线粒体动力学 线粒体自噬 

分 类 号：R285.5[医药卫生—中药学]