摘要
目的:探讨大鼠急性脑缺血再灌注后脑组织中细胞间黏附分子1的表达与白细胞浸润的关系,观察中药川芎嗪的治疗作用,并进行不同时限点的分析。方法:实验于2002-09/2003-02在湖南中医学院中心实验室进行。①分组:健康SD大鼠125只随机分为假手术组,模型组和川芎嗪组3组,假手术组分术后3,6,12,24和48h组,后两组又分为缺血3,6,12,24和48h组和再灌注3,6,12,24和48h组,每个时相点5只动物。②造模:假手术组仅分离动脉,不插入线栓,术后即刻腹腔注射生理盐水8mL/kg,其他两组线栓法制备脑缺血或缺血再灌注模型。③给药:川芎嗪组于缺血或再灌注后腹腔注射川芎嗪50mg/kg(71g/L),24h组追加给药1次,48h组追加给药两次。假手术组和模型组腹腔注射生理盐水8mL/kg。④指标检测:大鼠按相应时相点麻醉后断头处死,取脑组织免疫组化测定大鼠脑内细胞间黏附分子1表达阳性微血管数,苏木精-伊红染色观察白细胞计数,并分析两者间的相关性。结果:经补充后125只大鼠进入结果分析。①脑内细胞间黏附分子1表达阳性微血管数:免疫组化显示假手术组两侧半球大脑皮质可见少量表达。与假手术组比,模型组缺血6h表达明显增多,持续至48h时仍维持较高水平(P<0.01),再灌注组3h即明显高于对照组,24h达高峰(P<0.01)。川芎嗪组缺血及缺血再灌注6,12,24和48h均明显低于模型组相应时间点(P<0.01)。②白细胞计数结果:苏木精-伊红染色显示假手术组术后3h可见脑内针道周围有极少量,以后未继续增加。模型组脑缺血后3h即见,以后进行性增加,缺血再灌注3h有少量,6h逐渐增多,24h达到高峰,均显著高于对照组(P<0.01)。川芎嗪组缺血及缺血再灌注6,12,24和48h均明显低于模型组相应时间点(P<0.01)。③细胞间黏附分子1表达与白细胞浸润的相关性:脑缺血再灌注时两者呈正相关(rI=0.854,rIR=0.825,P<0.01)。结论:脑缺血再灌注后细胞间黏附分子1的表达与白细胞浸润在时相上有相关性,细胞间黏附分子1表达略早于白细胞浸润,提示细胞间黏附分子1可介导白细胞和内皮细胞的黏附。川芎嗪在不同时限点均可显著下调细胞间黏附分子1的表达,从而进一步抑制白细胞浸润,减轻缺血脑组织炎症反应。
AIM： To explore the relation between the expression of intercellular adhesion molecule-1 （ICAM-1） and leukocyte infiltration in rats after acute ischemia reperfusion, observe the therapeutical effect of traditional Chinese medicine ligustrazien, and analyze among the different time points. METHODS： The experiment was performed from September 2002 to February 2003 in Center Laboratory of Hunan College of Traditional Chinese Medicine. ① Divide into groups： 125 healthy SD rats were divided randomly into 3 groups, including sham operation group, model group and ligustrazine group. Sham operation group included post-operation 3, 6, 12, 24 and 48 hours groups, the latter＇two groups included ischernia 3, 6, 12, 24 and 48 hours groups and reperfusion 3, 6, 12, 24 and 48 hours groups with 5 animals in every time point. ② Build model： the rats in sham operation group only separated artery without inserted thread occlusion, and after operation 8 mL/kg saline was injected into abdominal cavity at once. In other two groups cerebral ischemia or ischemia reperfusion model was made by using thread occlusion. ③ Administration： The rats in ligustrazine group was injected with 50 mg/kg（71 g/L） ligustrazine in abdominal cavity after ischemia or reperfusion at the 24^th hour additional administration once and at 48^th hour additional administration twice. The saline 8 mL/kg injected in the abdominal cavity of rats in sham operation group and model group. ④ The detection of index： The rats were cut the heads to sentence to death after anesthesia based at the corresponding time point to gain brain tissue and detected the positive microvessel count （MVC） of ICAM-1 expression with immunohistochemistry. Using hematoxylin and eosin （HE） staining observed the numeration of leukocyte and analyzed the correlation between them. RESULTS： 125 rats were involved in the result analysis after compensation. ① The positive MVC of ICAM-1 expression： The immunohistochemistry showed that there was a little expression could be seen on cerebral cortes of amphi-hemisphere in sham operation group. Compared with sham operation group, the expression in model group after 6 hours ischemia significant increase, lasting to the 48^th hours it still maintained a high level （P〈 0.01）, at 3^nd hour reperfusion it was significantly higher than control group and reached the peak at 24^th hour （P 〈 0.01 ）. It was significantly lower of the rats with ischemia in ligustrazine group and at ischemia reperfusion 6, 12, 24 and 48 hours than that of model group at the corresponding time point （P 〈 0.01 ）. ② The result of numeration of leukocyte： the HE staining showed that it appeared a little around needle passage in brain 3 hours post-operation in sham operation group, and without continuously increased afterward. It was seen 3 hours after ischemia in model group, and ongoingly increased afterward, appearing a little at 3^nd hour ischemia reperfusion, and increased gradually at 6^th hour, and reached the peak at 24^th hour, which were all significantly higher than that in control group （P 〈 0.01 ）, It was significantly lower in ligustrazine ischemia and ischemia reperfusion 6, 12, 24 and 48 hours than that in model group at corresponding time point （P 〈 0.01 ）. ③ The correlation between the expression of ICAM-1 and leukocytic infiltrate： They were positive correlation at isehemia reperfusion （rI=0.854, rIR=0.825, P 〈 0.01）. CONCLUSION： The expression of ICAM-1 and leukocytic infiltrate has correlation at time points after cerebral ischemia reprfusion. The expression of ICAM-1 was a little earlier than that of leukocytic infiltrate, which indicates that ICAM-1 can mediate the adhesion of leucocyte and endothelial cell. The expression of ICAM-1 can be decreased significantly by ligustrazine at different time point in order to inhibit the leukocytic infiltrate further, and reduce inflammatory reaction of ischemia brain tissue.
出处
《中国临床康复》
CSCD
北大核心
2005年第25期149-151,共3页
Chinese Journal of Clinical Rehabilitation
基金
湖南省自然科学基金资助项目(99JJy1007)~~
