中药抗脑缺血再灌注损伤机制及超微结构研究

目的:探索单剂中药组合对脑缺血/再灌注损伤机理。方法:Wistar雄性大鼠随机分为4组:中药组、尼膜通组、对照组及手术对照组。线栓法制作大鼠大脑中动脉可逆性脑缺血模型。再通前20min经尾静脉中药组注入丹参、川穹嗪和当归;尼膜通组注入尼膜通;对照组给予生理盐水。再通后12、24和36h重复给药。再通48h,提取分离白细胞,测定线粒体超氧化物歧化酶(SOD)活性;采用Western印迹法,测定鼠抗羊单克隆抗体(Calbidin)和鼠抗羊单克隆抗体(LFA-1)表达水平;电镜下观察超微结构。结果:中药组大鼠缺血区脑组织线粒体中SOD活性及Calbindin表达水平最高。而中药组LFA-1表达水平却最低。电镜结果表明,中药组缺血神经元、神经胶质细胞、血管内皮细胞损害及炎性细胞的血管黏附均最弱。结论:单剂中药组合通过抗炎、清除自由基、钙离子拮抗机制对脑缺血/再灌注损伤起到保护作用。

Study of Mechanism and Ultrastructure of Chinese Traditional Medicine Against Cerebral Ischemic/Reperfusion Injury

Objective:To explore of mechanism of the composition of Chinese traditional medicinal monocase against cerebral ischemic/reperfusion injury.Methods:Wistar male rats were randomly divided into 4 groups:Chinese traditional medicine group,Nimotop group,control group and operation control group.Middle cerebral artery(MCA)were embolized by inserting a silicone embolus to establish reversible cerebral ischemic/reperfusion models.Before reperfusion 20 minutes,the rats of Chinese traditional medicine group,Nimotop group and control group were respctiveing injected Radix Salviae Miltiorrhizae + Ligustrazine + Angelic.Nimotop,and same quantitative 0.9% sodium choride through trail vein.After reperfusion 12,24 and 36 hours,the medicines were injected again by same way.After reperfusion 48 hours,leucocytes were separated;the activity of SOD was measured;the expressive level of Calbidin and LFA-1 by Western spot;The electrmicroscope was used to observe their ultrastructure.Results:The activity of SOD and the expression of calcium combining protein Calbidin in Chinese traditional medicine group was highest in three groups.However adhesive molecule LFA-1 on leucocytes was lowest.The electromicroscope result showed the injury extent of ischemic neuron,glial,microvessel and inflammatory also were lowest in three groups.Conclusion:The composition of Chinese traditional medicinal monocase depending on the effects of cleaning free radical scavenging,calcium antagonizing,inflammatory inhibition is against cerebral ischemic/reperfusion injury.