目的:研究中药胃肠舒对糖尿病(diabetes mellitus,DM)大鼠胃窦平滑肌细胞线粒体形态、结构、功能和蛋白的影响,探讨胃肠舒促胃肠运动的线粒体动能调控机制。方法:选择体重150~170 g的SD大鼠随机分为4组:正常对照组、DM组、DM+胃肠舒Ⅰ组(750 mg·kg^-1)、DM+胃肠舒Ⅱ组(1 500 mg·kg^-1)。透射电子显微镜检测线粒体形态、结构的变化,流式细胞术检测线粒体跨膜电位的变化,荧光测定法检测线粒体呼吸链的变化,Western Blot检测线粒体蛋白的变化。结果:胃肠舒能改善DM大鼠胃窦平滑肌细胞线粒体的形态、结构,增强呼吸链功能,线粒体跨膜电位和ATP含量增高,细胞色素C氧化酶(cytochromeC oxidase,COX)活性增强,活性氧自由基(ROS)释放减少,凋亡蛋白家族Bcl-2表达上调,Bax表达下调。结论:胃肠舒促胃肠运动可能与改善平滑肌的线粒体动能系统有关。
Objective:To study the mechanism of the Chinese medicine Weichangshu on mitochondrial kinetic energy of stomach in diabetic mellitus(DM) rats by measuring the changes in morphology,structure,function and protein of mitochondria.Method:Standard Sprague Dawley rats(150-170 g) were randomly divided into 4 groups,including a control group,DM model group,DM + WeichangshuⅠgroup(750 mg·kg^-1),DM + Weichangshu Ⅱ group(1 500 mg·kg^-1).The mitochondrial morphology and structure were detected using transmission electron microscopy.The changes in mitochondrial transmembrane potential,respiratory chain and mitochondrial proteins were detected using flow cytometry,fluorescence labeling and Western blot respectively.Result:Weichangshu could ameliorate the mitochondrial morphology,structure and respiratory chain of stomach in diabetic mellitus rats.Mitochondrial transmembrane potential,ATP level and activity of cytochrome C oxidase were increased.The release of ROS was decreased.The expression of Bcl-2 was up-regulated and the expression of bax was down-regulated.Conclusion:Weichangshu could promote the gastrointestinal motility,which might be related with the regulation of mitochondrial kinetic energy.