转基因神经干细胞移植在缓解脑缺血性神经血管损害中的作用  

Amelioration of neural and vascular damage by transplantation of gene-transfected neural stem cells after transient cerebral ischemia

作　　者：朱巍[1] 毛颖[1] 周良辅[1] 汪洋[2] 江永[1] 朱剑虹[1] 

机构地区：[1]复旦大学附属华山医院神经外科,上海200040 [2]复旦大学上海医学院解剖组织胚胎系

出　　处：《中华实验外科杂志》2005年第2期198-199,i002,共3页Chinese Journal of Experimental Surgery

基　　金：上海市科技发展基金重点资助项目 (0 2DJ1 4 0 2 5)

摘　　要：目的　评价转染血管内皮生长因子 (VEGF)基因的神经干细胞脑内移植后在宿主缺血区的神经血管保护作用。方法　建立大鼠暂时性大脑中动脉梗死模型 (tMCAO)并将其随机分成 :(1)对照组 ;(2 )细胞悬液磷酸盐缓冲液 (PBS)移植组 ;(3 )神经干细胞移植组 ;(4 )转基因神经干细胞移植组 ,每组 10例。立体定向法将BrdU标记的转基因神经干细胞移植到tMCAO大鼠的纹状体缺血半暗区。移植后 2～ 12周进行神经损害严重程度评分 (NSS)并与其他 3组比较。移植后12周 ,免疫荧光染色观察转基因神经干细胞在脑内的分化、迁徙情况 ;免疫组织化学染色观察各组移植区毛细血管结构 ,并计每例动物以移植点为中心的一个 10 0倍视野下内皮细胞数。结果　移植后 2～ 12周 (4 )组NSS评分均数分别为 5 .8、5 .0、4.6、4.0、4.0、3 .8,均低于其他 3组。其中第 8周显著低于 (1)、(2 )组 (P =0 .0 0 8) ,第 12周显著低于 (1)、(2 )、(3 )组 (P =0 .0 0 0 )。移植后 12周 ,转基因神经干细胞在宿主脑内存活、迁徙 ,部分分化成神经元。转基因神经干细胞移植组移植区毛细血管结构相对完整 ,血管内皮细胞数显著多于其他 3组。结论　转染VEGF基因的神经干细胞移植后对宿主局部血管和神经结构具有保护作用。Objective To evaluate the effectiveness of VEGF-transfected neural stem cells (NSCs) in protection of vascular and neural tissue in rat brain subjected to ischemia.Methods Temporary middle cerebral artery occlusion (tMCAO) models were established and randomly divided into (1) control group (n=10), (2) PBS transplantation group (n=10),(3) neural stem cells transplantation group (n=10) and (4) VEGF-secreting neural stem cells transplantation group (n=10).BrdU-labelled VEGF-secreting NSCs were transplanted into the penumbra zones respectively 3 days after tMCAO model established.NSS was checked in all groups 2,4,6,8,10,12 weeks respectively after transplantation.By using immunofluorescent staining,differentiation and migration of transplanted neural stem cells 12 week after transplantation were detected.The number of endothelial cells around the transplantation area was measured by immunohistochemical staining.Results NSS in group 4 was 5.80± 1.48, 5.00± 1.05, 4.60± 0.97, 4.00± 0.67, 4.00± 1.05 and 3.80± 0.42 respectively from 2-12 weeks after transplantation,significantly lower than those in groups 1 and 2 at 8th week (P= 0.008) and those in groups 1,2 and 3 at 12th week (P=0.000) after transplantation.Twelve weeks after transplantation,transplanted VEGF-secreting NSCs survived and migrated,and some of them differentiated to neurons and integrated well with hosts' cytoarchitectural components.The number of endothelial cells in group 4 was significantly more than that in other 3 groups.Conclusion VEGF-transfected NSCs can reduce brain injury through protecting the vascular system against ischemia and reperfusion injury.

关 键 词：神经干细胞移植 移植后 神经血管 脑内 血性 毛细血管 脑缺血 转基因 迁徙 宿主 

分 类 号：R651[医药卫生—外科学]