COX-2抑制剂对幼鼠反复痫性发作后神经发生的影响

目的：研究环氧合酶-2（COX-2）在痫性发作活化后的表达特点，探讨COX-2抑制剂塞莱昔布（eeleeoxib，Cel）对痫性活动后海马区神经发生的影响。方法：模型制作：随机将120只体重为50～60g的3周龄健康Wistar幼鼠分为匹鲁卡品致痫组（EPOnly组）（n=45）和Cel干预致痫组（EP—Cel）（n=45）和生理盐水正常对照组（NS组）（n=30）3组。随机在EP—only及EP—Cel组各取10只匹鲁卡品成功诱导急性发作幼鼠进行腹腔注射溴脱氧尿核苷（BrdU）：（1）行为学观察：根据Racine分级评价急性期和慢性期痴性发作行为；（2）形态学检测：各实验组分别在急性发作后第14天，第28天处死大鼠制备组织切片进行免疫组化检测COX-2阳性细胞在各组的表达变化趋势，以及BrdU＋神经元特异性核蛋白（NeuN）和BrdU＋星形胶质细胞胶原纤维酸性蛋白（GFAP）荧光免疫双标阳性细胞的表达，观察神经前体细胞的增殖及分化在各组的差异。结果：（1）动物行为学观察：在急性期，EP-only组全身性自发反复性癫痫发作（SRS）发作率（90％）明显高于EP—Cel组（560）（P〈0．01）；EP—only组痫性发作Racine分级强度（3．7±1．3）明显高于EP—Cel组发作强度（2．5±1．1）（P〈0．05）；在慢性期，EP—Only组SRS发生率（500）明显高于EP—Cel组（30％）（P〈0．05；岔检验）；EP—Only组平均每天SRS发生的频率（1．9±0．58）明显高于EP=Cel组（0．6±0．3）（P〈0．01，t检验）；（2）形态学检测免疫组化结果：①COX-2免疫反应阳性细胞的表达：匹鲁卡品致痫第14天后，海马区COX-2阳性细胞表达在EP—Only组明显高于EP-Cel组[（158±18）vs（118±20）]（P〈0．01）；②BrdU＋NeuN和BrdU＋GFAP免疫双标结果：急性期发作第28天后，BrdU＋NeuN免疫双标阳性细胞在EP-Only组明显高于EP—Cel组[（36±4）vs（22±3）]；同时EP-Only组门区有BrdU＋GFAP免疫双标阳性的新生的胶质细胞明显比EP—Cel组高[（26±3）vs（14±2）3。结论：COX-2在痂性发作后被迅速诱导表达，COX-2抑制剂Cel能抑制痫性发作激活的异常神经发生和星形胶质细胞增生，减少慢性期SRS。

The effect of COX-2 inhibitor on neurogenesis afte recurrent sei-zures in immature rats

Objective： To study the expression of COX-2 after epileptic seizures and the effect of COX-2 inhibitors on neurogenesis in the immature epileptic brain. Methods： Three-week-old Wistar rats （n： 120） were randomly divided into three groups： the pilocarpine-induced epilepsy group （EP-Onty group,n=45）, Celecoxib treatment epilepsy group （EP-Cel group,n=45）, and the normal saline control group （NS,n=30）. Pilocarpine induced status epileptieus model. Seizures were scoreded by Racine＇s scale. （1） Animal behavior observation： to compare the difference in acute phase of SE and monitoring spontaneous recurrent seizure （SRS） in chronic phase from 28th to 42rd days after SE; （2） Morphological Detection： The immunoreactive cells were detected by immunohistochemistry and histology, in the 3 groups. The detected indexes were as follows： COX-2, BrdU and NeuN doubled stained to mark newly generated neurons, and BrdU and GFAP doubled stained to mark newly generated astroglls. Results： （1） Observation of animal behavior showed：① the acute phase： seizure rate in EP-only group （90%） was significantly higher than in EP-Cel group （56 % ） （P 〈0.01 ）;②Chronic phase monitoring： Incidence of SRS in EP-Only group （50%）was significantly higher than that in EP-Cel group （30%）（P〈0.05;χ^2 test）; （2）Iimmunohistoehemieal results： Ⅰ COX-2-positive cells in the hippocampus of EP-only group was significantly higher than that in El＇ Celecoxib group[（158 ± 18）vs（118±20） 1 （P〈0. 01）. The re subs of double immunofluorescence：28 days after acute attack, BrdU ＋ NeuN immune double positive cells in EP Only group was significantly higher than that of EP Cel group[（36 ± 4）vs（22 ± 3）]; at the samc time BrdU 5 GFAP double stained positive cells in EP-Only group was significantly higher than that of EP-Cel group[（26 ± 3）vs（14 ± 2）]. Conclusions：The activation of COX-2 can be induced rapidly after seizures, and COX 2 inhibitor attenuated the likelihood of developing spontaneous recurrent sei zures,and prevented ectopic neurogenesis in the hilus .