通心络对大鼠脑缺血再灌注模型微血管细胞间黏附分子1和血管细胞黏附分子1表达的影响

摘要

背景:现代医学发现通心络制剂除了具有抗凝和抑制血小板聚集作用外,对血管内皮细胞有一定的保护作用.目的:观察中药复方制剂通心络是否影响脑缺血再灌注动物模型黏附分子的表达.设计:随机对照实验.单位:解放军第二军医大学长征医院神经内科.材料:实验于2002-10/2003-01在解放军第二军医大学长征医院神经内科实验室完成.选择雄性SD大鼠25只,随机分为假手术组5只、模型组10只和通心络组10只.方法:线栓法制备大鼠大脑中动脉脑局灶性脑缺血再灌注模型,假手术组除将尼龙线插在颈外动脉接近颈内动脉分叉处外,其余同模型组.通心络组大鼠在缺血再灌注前给予通心络粉剂1.0 g/(kg·d),溶在生理盐水中灌胃1周.模型组和假手术组灌胃等剂量生理盐水.各组大鼠麻醉后取脑制备切片,行常规苏木精-伊红染色、免疫组化及原位杂交染色.主要观察指标:①缺血再灌注后细胞间黏附分子1和血管细胞黏附分子1阳性微血管表达数目.②缺血再灌注后细胞间黏附分子1 mRNA阳性微血管表达数目.结果:①假手术组手术侧大脑半球皮质和基底节区未见细胞间黏附分子1、血管细胞黏附分子1蛋白和细胞间黏附分子1 mRNA阳性微血管表达.②模型组大鼠缺血2 h再灌注6 h后,缺血侧大脑细胞间黏附分子1、血管细胞黏附分子-1蛋白表达水平和细胞间黏附分子1 mRNA表达水平显著升高.③通心络组缺血侧大脑半球皮质和基底节区蛋白和mRNA阳性微血管数较模型组显著降低[(10.42±1.98),(12.42±2.14)/高倍视野;(8.54±2.00),(11.12±1.56)/高倍视野](P＜0.05),血管细胞黏附分子1蛋白阳性微血管表达数目无显著变化(P＞0.05).结论:通心络可以降低大鼠脑缺血再灌注后细胞间黏附分子1的转录和翻译过程,有助于减轻脑缺血后的炎症性损伤过程.%BACKGROUND: Apart from anticoagulation property and suppressing platelet congregation capability, tongxinluo preparation has been proved by traditional Chinese medicine to possess certain function for protecting endothelial cells.OBJECTIVE: To observe the influence of Chinese medicinal herb "tongxinluo" compound on adhesion molecule expression in brain ischemia-reperfusion (IR) animal model.DESIGN: Randomized and controlled experiment.SETTING: Department of Neurology, Changzheng Hospital Affiliated to the Second Military Medical University of Chinese PLA.MATERIALS: This experiment was conducted at the laboratory of the Department of Neurology, Shanghai Changzheng Hospital, between October 2002 and January 2003. Totally 25 male SD rats were randomized into sham-operation group of 5 rats, model group of 10 rats and tongxinluo group of 10 rats.METHODS: Middle cerebral artery was occluded using thread-bolt method to induce focal brain IR model in rats. In sham-operation group,nylon thread was placed around the external carotid artery approximating to the branch of internal carotid artery, and the other procedure was the same as that in model group. Rats in tongxinluo group were given tongxininfusion before IR for 1 consecutive week, which was replaced by physiological saline of the same dosage in model group and sham-operation group. Brain tissues were obtained under anesthesia condition and cut into slices; conventional HE staining, immunohistochemical and in situ hybridization staining were conducted.MAIN OUTCOME MEASURES:① The number of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1)positive microvessels following IR injury.② The number of ICAM-1 and VCAM-1 mRNA positive microvessels following IR injury.RESULTS:① In sham-operation group,ICAM-1,VCAM-1 protein andICAM-1 mRNA positive microvessels could not be observed in hemispheric cortex and basal ganglion at the operative side.② In model group,the positive expression of ICAM-1, VCAM-1 protein and ICAM-1 mRNA obviously increased at the ischemic side at 6-hour reperfusion following 2-hour ischemia.③ In tongxinluo medication group,the positive protein and mRNA-expressing microvessls were found remarkably reduced in number in ischemic side hemispheric cortex and basal ganglion [(10.42 ±1.98),(12.42±2.14)/HP; (8.54±2.00), (11.12±1.56)/HP] (P ＜ 0.05), but the positive VCAM-1 protein-expressing microvessels did not change remarkably (P ＞ 0.05).CONCLUSION: Tongxinluo can suppress ICAM-1 transcription and translation following rat brain IR, thus attenuating inflammatory injury induced by brain ischemia.