小续命汤有效成分组对脑缺血/再灌注大鼠恢复早期脑线粒体的保护作用研究  

Effects of active components group of Xiaoxuming decoction on brain mitochondria in cerebral ischemia/reperfusion rats during early recovery period

作　　者：杜肖[1] 路畅[1] 贺晓丽[1] 杜冠华[2] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]中国医学科学院北京协和医学院药物研究所,北京100050

出　　处：《中国中药杂志》2017年第11期2139-2145,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81473586;81202192;81273654)

摘　　要：观察小续命汤有效成分组对脑缺血/再灌注大鼠恢复早期脑线粒体的作用,研究其对脑缺血/再灌注大鼠恢复早期的神经保护作用机制。采取插线法制备大鼠大脑中动脉阻塞脑缺血模型,2 h后再灌注。应用Zea-Longa's级标准评分法评价动物脑缺血程度,将模型成功大鼠随机分为模型组、小续命汤有效成分低、中、高剂量组和阳性药金纳多组,以假手术大鼠作为对照组。于给药5 d后梯度离心提取大鼠脑组织缺血周边区线粒体,通过Clark氧电极法检测线粒体呼吸功能,荧光探针法检测线粒体内ROS含量及线粒体膜电位,分光光度法检测线粒体琥珀酸脱氢酶活性和脑组织ATP含量。结果显示,小续命汤有效成分组能够显著改善脑缺血再灌注大鼠恢复早期脑线粒体的呼吸功能,提高线粒体琥珀酸脱氢酶的活性,降低线粒体ROS的含量,提高脑线粒体膜电位,促进脑组织ATP的合成。提示小续命汤有效成分组能够明显减轻脑缺血再灌注早期导致的脑组织能量代谢紊乱,改善脑线粒体结构和功能损伤,说明小续命汤有效成分组对脑线粒体的保护作用可能是其发挥神经保护的作用机制之一。To observe the effect of active components group of Xiaoxuming decoction （XXMD） on brain mitochondria in cerebral ischemia/reperfusion rats during early recovery period, and study its protective mechanism for nerves in cerebral ischemia/reperfusion rats during early recovery period. Cerebral ischemia model of middle cerebral artery occlusion in rats was established by suture method, and reperfusion was conducted 2 h later. The degree of cerebral ischemia in rats was evaluated by using Zea-Longa＇s standard grading method, and the model rats were randomly divided into model group, Xiaoxuming decoction active components low, medium and high dose groups and positive drug Ginaton group, with sham operated rats as control group. Gradient centrifugation was used to extract the mitochondria from rat brain after 5 days of drug administration. Then the mitochondrial respiratory function was measured by Clark oxygen electrode method; mitochondrial membrane potential and the mitochondrial reactive oxygen species （ROS） level were detected by fluorescence probe methods; and the activity of mitochondrial succinodehydrogenase （SDH） and the content of ATP in the ischemic re- gion of MCAO rats were measured by spectrophotometric method. The results showed that as compared with the model group, XXMD could significantly improve mitochondrial respiratory activity, increase the activity of SDH, reduce the level of ROS, increase mitochon- drial membrane potential and obviously promote the synthesis of ATP in brain tissues. The results indicated that XXMD active compo- nents group could alleviate the energy metabolism disorders, protect brain mitochondrial damage and improve mitochondrial function in MCAO rats, which may be the mechanism of its neuroprotection activity.

关 键 词：小续命汤有效成分组 脑缺血/再灌注 线粒体功能 氧化磷酸化 活性氧 膜电位 

分 类 号：R285.5[医药卫生—中药学]