大鼠脑缺血再灌注后细胞间黏附分子1表达与白细胞浸润的关系及吲哚美辛的干预效应

Relationship between the expression of intercellular adhesion molecule-1 and leucocytic infiltration after cerebral ischemical reperfusion in rat as well as the interventional effect of indomethacin

目的:观察脑缺血2h再灌注不同时间点脑组织细胞间黏附分子1的表达与白细胞浸润的关系,并观察吲哚美辛对细胞间黏附分子1表达及白细胞浸润的影响方法:实验于2004-09/2005-03在大连医科大学附属第二医院中心实验室完成。取35只雄性SD大鼠随机分为假手术组5只、对照组25只,吲哚美辛组5只,对照组又分为缺血2h再灌注2,12,24,48,72h5个亚组,每个亚组5只。①造模:采用线栓法制作大鼠大脑中动脉阻断模型,阻断大鼠大脑中动脉血流2h之后进行再灌注。②给药:吲哚美辛组在再灌注24h时灌胃给予吲哚美辛(按10mg/kg,溶于2mL生理盐水中),对照组中的再灌注24h亚组给予等容生理盐水。③观察指标:经免疫组化和苏木精-伊红染色,测定细胞间黏附分子1表达阳性微血管数和白细胞计数。结果:35只大鼠均进入结果分析。①在脑缺血再灌注早期坏死区周围微血管内皮细胞表达细胞间黏附分子1即开始增多[再灌注2h:(15.94±1.90)个/视野,再灌注12h:(30.73±3.01)个/视野],并于24h达高峰[(37.86±2.21)个/视野],各对照亚组与假手术组[(0.68±0.69)个/视野]比较差异有显著性意义(P<0.01),且各亚组间比较差异有显著性意义(P<0.01)。②在脑缺血再灌注早期坏死区周围白细胞浸润即开始增多[再灌注2h:(2.30±0.91)个/视野,再灌注12h:(9.99±1.40)个/视野],并于24h达高峰[(22.11±1.71)个/视野],各对照组亚组与假手术组比较有显著性差异(P<0.01),且各亚组两两比较组间差异有显著性意义(P<0.01)。③细胞间黏附分子1表达与白细胞浸润的相关性分析表明两者之间呈正相关(r=0.731,P<0.01)。④吲哚美辛组细胞间黏附分子1表达及白细胞浸润计数[(16.01±11.43)个/视野,(10.55±2.64)个/视野]均低于再灌注24h亚组(P<0.01)。结论:脑缺血再灌注后细胞间黏附分子1可介导白细胞与内皮细胞的黏附;吲哚美辛可降低脑缺血再灌注后细胞间黏附分子1的表达和白细胞的浸润,对脑缺血再灌注损伤具有保护作用。

AIM：To observe the relationship between the expression of intercellular adhesion molecule-1（ICAM-1） and leukocytic infiltration in brain tissue at different time points of reperfusion 2 hours after cerebral ischemia,and observe the effect of indomethacin on the expression of ICAM-1 and leukacytic infiltration. METHODS：This experiment was conducted in the Central Laboratory of Second Affiliated Hospital of Dalian Medical University between September 2004 and March 2005.Totally 35 male Sprague-Dawley （SD） rats were randomly divided into pseudo operation group（n=5）, control group（n=25） and indomethacin group （n=5）.Control group was divided into ischemia 2 hours reperfusion 2, 12, 24, 48 and 72 hours 5 subgroups. ①Making models： Making occlusion model of middle cerebral artery in rat with line embolism method,reperfusion was executed 2 hours after middle cerebral artery of rat was occluded. ②Administration：Indomethacin was used 24 hours after reperfusion by intragastric administration to indomethacin group（according to 10 mg/kg,dissolved into 2 mL of normal saline）, isasteric normal saline was given to 24 hours subgroup of control group. ③Observed index： Measuring the number of microvessel whose expression of ICAM-1 was positive and leucocyte count after being stained with immunohistachemical method and hamatoxylin-eosin. RESULTS-Totally 35 rats entered the result analysis. ④Increase of expression of ICAM-1 in microvascular endothelial cell around the zone of necrosis began in early stage of cerebral ischmical reperfusion[reperfusion 2 hours： （15.94±1.90）/field,reperfusion 12 hours： （30.73±3.01）/field], and reach the peak at hour 24 [（37.86±2.21 ）/field] ,there was signfflcant difference between every subgroup of control group and pseudo operation group [（0.68±0.69）/field,P 〈 0.01] and there was significant difference between each two subgroups（P 〈 0.01）;②Leukocytic infiltration around the zone of necrosis in early stage of cerebral isehmical reperfusion began to increase [reperfusion 2 hours.- （2.30±0.91 ）/field ,reperfusion 12 hours： （9.99±1.40）/field], and reached the peak at hour 24[（22.11±1.71 ）/field], there was significant difference between every subgroup of control group and pseudo operation group （P 〈 0.01）and there was significant difference between each two subgroups （P〈 0.01）; ③Relativity analysis of the expression of ICAM-1 and leukocytic infiltration showed the positive correlation（r=-0.731,P 〈 0.01 ）.④The expression of ICAM-1 and leucocytic infiltration count in the indomethacin group [（16.01±11.43）/field, （10.55±2.64）/field]was both lower than that in the reperfusion 24 hours subgroups （P 〈 0.01）. CONCLUSION：ICAM-1 could mediate adhesion between leucocyte and endotheliocyte; Indomethacin can decrease the expression of ICAM-1 and leucocytic infiltration after cerebral ischemical reperfusion and was of protective effect on cerebral ischemical reperfusion injury.