脑缺血再灌注损伤后炎症反应与赛来昔布的脑保护作用

Brain protection of celebrex and the inflammatory reaction following cerebral ischemia-reperfusion injury

目的:细胞间黏附分子1及白细胞浸润参与了脑缺血再灌注损害过程,观察赛来昔布对局灶性脑缺血再灌注损伤大鼠的脑梗死体积、脑缺血坏死区周边细胞间黏附分子1和白细胞浸润的影响,探讨其是否具有脑保护作用。方法:实验于2003-12/2004-02在大连医科大学附属第二医院中心实验室进行。SD大鼠36只随机分为3组:赛来昔布组(n=16)、安慰剂组(n=16)及假手术组(n=4),其中赛来昔布组和安慰剂组按脑缺血再灌注2,4,6,24h分为4个亚组(n=4)。手术前10min赛来昔布组用赛来昔布灌胃(0.25mg/g,以3mL生理盐水溶解),安慰剂组安慰剂灌胃,剂量相同,然后参照改良的Longa-Zea线栓法制作大脑中动脉闭塞及再通模型,假手术组仅做颈部正中切口暴露右侧颈总动脉后缝合皮肤。观察大鼠大脑中动脉闭塞2h再灌注2,4,6,24h4个时相点梗死体积,缺血区白细胞计数,并应用免疫组织化学法染色观察细胞间黏附分子1阳性微血管数。结果:36只大鼠进入结果分析。①赛来昔布组再灌注后2,4,6,24h的梗死体积明显小于同时段安慰剂组[(4.81±0.13)%,(8.22±0.25)%,(11.83±0.62)%,(15.93±0.42)%,(6.56±2.07)%,(10.12±2.37)%,(13.53±1.47)%,(17.86±2.78)%,P<0.05]。②假手术组细胞间黏附分子1阳性表达为(0.63±0.62)个,未见白细胞浸润。脑缺血再灌注后,脑缺血坏死区周边微血管内皮细胞细胞间黏附分子1达及白细胞浸润增多,并于24h达高峰,此时细胞间黏附分子1表达及白细胞浸润赛来昔布组低于同期安慰剂组[(43.00±1.41),(6.25±1.26),(59.50±2.25),(6.75±1.8)个,P<0.01]。结论:赛来昔布可缩小脑缺血再灌注后的脑梗死体积,抑制局灶性脑缺血再灌注时炎症反应,非特异性地起到脑保护作用。

AIM:To study the effect of pretreatment with celebrex on volum of cerebral infarction, expression of intercellular adhesion molecule 1 and leukocytic infiltrate in rats with focal ischemiareperfusion injury so as to study the effect on brain protection.METHODS:The experiment was performed in the Central Laboratory,Second Affiliated Hospital of Dalian Medical University from December 2003 to February 2004.A total of 36 male SD rats were divided randomly into celebrex group(n=16),placebo group(n=16),and sham operation group(n=4). Celebrex and placebo groups were subdivided into 2,4,6and 24hour groups(n=4) according to ischemiareperfusion time.Rats in celebrex group received celebrex(0.25 mg/g diluted with 3 mL saline),and placebo group received the same dose of placebo 10 minutes before operation to establish models of middle cerebral artery occlusion and reperfusion with LongaZea thread occlusion.Rats in sham operation group received cervical median incision to expose right common carotid artery,and then the skin was sutured.The volume of cerebral infarction in rats at reperfusion for 2,4,6 and 24 hours following occlusion for 2 hours and the amount of leukocytes in ischemic area were observed.The positive expression of intercellular adhesion molecule 1 was observed with immunohistochemical method.RESULTS:Totally 36 rats were involved in the result analysis.①The volume of infarction after reperfusion for 2,4,6,and 24 hours in celebrex group[(4.81±0.13)%,(8.22±0.25)%,(11.83±0.62)%,(15.93±0.42)%] was obviously smaller as compared with that in placebo group[(6.56 ±2.07)%,(10.12±2.37)%,(13.53±1.47)%,(17.86±2.78)%,P< 0.05].②The positive expression of intercellular adhesion molecule 1 in sham operation group was 0.63±0.62,and no leukocytic infiltrate was showed.After cerebral ischemiareperfusion,the positive expression of intercellular adhesion molecule 1 in microvascular endothelial cells around necrosis and leukocytic infiltration increased,and reached to the peak at hour 24;at the same time,the positive expression of intercellular adhesion molecule1 and leukocytic infiltrate in celebrex group(43.00±1.41,6.25±1.26) were lower as compared with those in placebo group(59.50±2.25,6.75±1.8,P< 0.01). CONCLUSION:Celebrex can decrease the volume of cerebral infarction in rats with focal ischemiareperfusion injury,and inhibit the inflammatory reaction.Celebrex plays a role in brain protection.