背景:黄角颗粒为纯中药制剂,具有通腑泄热、开窍熄风、活血化淤等功效。目的:观察黄角颗粒对大鼠大脑中动脉栓塞模型神经细胞凋亡的影响,探讨黄角颗粒治疗急性期脑梗死的可能机制。设计:完全随机分组设计,对照动物实验。单位:华中科技大学同济医学院附属中西医结合医院神经内科。材料:实验于2003-09/2003-12在湖北中医学院中心实验室完成。采用鼠龄为22个月的老年Wistar大鼠60只。大鼠适应性喂养3d后,按体质量随机分为6组:正常对照组、模型组、尼莫地平组、黄角颗粒10g/kg组、黄角颗粒5g/kg组、黄角颗粒2.5g/kg组,每组10只。方法:①采用大脑中动脉栓塞法将大鼠造成脑缺血模型。黄角颗粒(主要成分为生大黄、水牛角等,武汉市第一医院制剂室提供,批号:021120)10,5,2.5g/kg组于造模前30min分别给予含生药1,0.5,0.25kg/L的黄角颗粒水溶液按10,5,2.5g/kg灌胃,每只灌胃1次;尼莫地平组于造模前30min给予含生药1g/L的尼莫地平水溶液按10mg/kg灌胃,每只灌胃1次;模型组和正常对照组(只结扎颈外动脉)于造模前30min给予生理盐水按10mL/kg灌胃,每只灌胃1次。②采用光学显微镜观察原位细胞凋亡染色切片,在高倍镜(20×10)下观察5个视野,观察每个高倍视野平均阳性细胞数,并计算细胞凋亡率。普通光镜下观察皮质缺血局部及周围组织学变化。电镜下观察大鼠大脑皮质细胞超微结构变化。③计量资料差异比较采用F检验及Q检验。主要观察指标:黄角颗粒对大脑中动脉栓塞老年大鼠脑组织神经细胞凋亡率及大脑皮质细胞形态学的影响。结果:Wistar大鼠60只均进入结果分析。①模型组大鼠脑组织凋亡阳性细胞数及细胞凋亡率明显高于正常对照组(P<0.01);黄角颗粒各剂量组及尼莫地平组大鼠脑组织凋亡阳性细胞数及细胞凋亡率明显低于模型组(P<0.01);黄角颗粒10g/kg组大鼠脑组织凋亡阳性细胞数及细胞凋亡率与尼莫地平组比较,差异不明显(P>0.05)。黄角颗粒10g/kg组大鼠脑组织凋亡阳性细胞数及细胞凋亡率明显低于黄角颗粒5和2.5g/kg组(P<0.05,0.01)。②光镜及电镜观察结果显示模型组大鼠局部脑组织细胞有明显的凋亡现象,而黄角颗粒有明显的改善作用。结论:黄角颗粒对急性缺血性神经细胞凋亡有一定的拮抗作用,该种作用可能是黄角颗粒治疗急性期脑梗死的可能机制之一。 BACKGROUND: Huangjiao granules are pure Chinese medicine preparations and have functions such as expelling heat, opening and extinguishing wind, promoting blood circulation, and removing stasis. OBJECTIVE: To observe the effect of Huangjiao granules on neuronal apoptosis in rat middle cerebral artery occlusion model and to explore the possible mechanism of Huangjiao granules in treatment of acute cerebral infarction. Design: Completely random grouping design, control animal experiments. Unit: Department of Neurology, Department of Integrated Traditional Chinese and Western Medicine, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: The experiment was performed at the Central Laboratory of Hubei University of Traditional Chinese Medicine from September 2003 to December 2003. 60 Wistar rats aged 22 months old were used. After adaptive feeding for 3 days, rats were randomly divided into 6 groups according to body weight: normal control group, model group, nimodipine group, Huangjiao granule 10g/kg group, Huangjiao granule 5g/kg group, Huangjiao granule 2.5g /kg group, 10 in each group. Methods: 1 The model of cerebral ischemia was induced by middle cerebral artery occlusion. Huangjiao granules (mainly composed of rhubarb, buffalo horn, etc., provided by the preparation room of the First Hospital of Wuhan City, batch number: 021120) 10, 5, 2.5g/kg group were given crude drug 1,0.5,0.25 respectively 30 minutes before modeling The kg/L solution of Huangjiao Granules was intragastrically administered at 10, 5, and 2.5 g/kg, and each stomach was given once; the nimodipine group was given 10 mg/L of Nimodipine aqueous solution containing 1 g/L of crude drug 30 minutes before modeling. The rats were intragastrically administrated by gavage with one gavage each time; the model group and normal control group (ligation of the external carotid artery only) were given intragastric administration of 10 mL/kg 30 min before modeling, and each stomach was intragastrically administered once. 2 Observe in situ apoptosis stained sections with light microscope. Observe 5 fields under high magnification (20×10). Observe the number of positive cells in each high power field and calculate the rate of apoptosis. Under normal light microscope, local and peripheral histological changes of cortical ischemia were observed. The ultrastructure changes of rat cerebral cortex were observed under electron microscope. 3 The differences in measurement data were compared using F-test and Q-test. MAIN OUTCOME MEASURES: The effect of Huangjiao granules on the neuronal apoptosis and the morphology of cerebral cortical cells in the middle cerebral artery occlusion of aged rats. Results: All 60 Wistar rats were involved in the analysis of the results. 1 The number of apoptotic cells and the apoptosis rate of brain tissue in the model group were significantly higher than those in the normal control group (P<0.01); the number of apoptotic positive cells in the brain of each group of Huangjiao granules and nimodipine group was The apoptosis rate was significantly lower in the model group than in the model group (P<0.01). The number of apoptotic cells in the brain and the apoptosis rate of the rat in the Huangjiao granule 10g/kg group were not significantly different from those in the nimodipine group (P>0.05). ). The number of apoptotic positive cells and the rate of apoptosis in the rat brain of Huangjiao granules in the 10g/kg group were significantly lower than those in Huangjiao granules group 5 and 2.5g/kg (P<0.05, 0.01). 2The results of light and electron microscopy showed that there was obvious apoptosis in the brain tissue of the model group, while the Huangjiao granules had obvious improvement. Conclusion: Huangjiao granules can antagonize apoptosis of acute ischemic neurons. This effect may be one of the possible mechanisms of Huangjiao granules in the treatment of acute cerebral infarction.