CT1修饰的神经干细胞移植癫痫持续状态大鼠海马后的迁移、分化及对空间记忆的影响

目的 观察心肌营养素1(CT1)修饰的神经干细胞(NSCs)移植癫痫持续状态(SE)大鼠海马后的存活、迁移和分化情况,观察对大鼠空间记忆功能的影响.方法 (1)将携带CT1基因的重组腺病毒(Adv-CT1)转染NSCs,并用增强型绿色荧光蛋白(EGFP)标记.(2)随机将54只SE大鼠分为联合移植组、单纯移植组及模型对照组(各18只),分移植后1、4及8周3个时间点(各6只).(3)共聚焦荧光显微镜下观察NSCs存活、分化及迁移;水迷宫试验观察空间记忆功能.结果 移植后4及8周,联合移植组双标阳性细胞数明显多于单纯移植组,并见细胞迁移,单纯移植组未见细胞迁移;联合移植组大鼠逃避潜伏期短于单纯移植组(P<0.05).结论 CT1能促进NSCs在SE大鼠脑内的存活、迁移和分化,NSCs-CT1联合移植可改善大鼠SE后空间记忆功能障碍.

Migration and differentiation of transplanting neural stem cells modified by cardiotrophin-1 in hippocampus and its effect on the spatial memory in status epilepticus rats

Objective To observe the survival,migration and differeraiation of grafted neural stem cells (NSCs) modified by cardiotrophin-1 (CT1)in hippocampus in status epilepticus (SE) rat,and its effect on the soatial memory in SE rats.Methods (1)NSCs were cultivated and transfected with adenovirus CT1,marked with enhance green fluorescent protein (EGFP) in vitro (2)54 Pilocarpine-SE model rals were divided into three groups randomly:CT1-NSCa-EGFP transplantation group(n=18);NSCs-EGFP trangplantation group (n=18);model control group(n=18).Another 18 rats served as normal control group,Each group was divided into three time points equally(each point,n=6)corresponding to 1 week,4 weeks and 8 weeks after transplantation.(3) Under the confocal microscopy,the survival,migration and differentiation of the grafted cells were observed by immunofluorescent stainirlg for EGFP(green),NF-200(red),and double-labeled(yellow).The spatial memory of the SE rate was tested by Morris wafer maze task. Results At 4 weeks and 8 weeks after transplanlation,the numbers of EGFP-positive and double-labeled cells in the CT1-NSCs-EGFP transplantation group were more significantly increased than those in the NSCs-EGFP transplantation group.In the CT1-NScs-EGFP transplantation group.mosl of the grafted NSCs migrated away from the needle tract.But in tlle NScs-EGFP transplantation group.the majority of grafted cells remained at the transplantation site.The latency period of escape markedly decreased at 4 and 8 weeks in the rats transplanted by CT1-NSCs-EGFP compared with those by NSCs-EGFP(P<0.05).Conclusions CT1 could promote the survival,migration and differentiation of engrafted NSCs.Engrafted NSCs combined with CT1 could improve the spatial memory in SE rats.