胶质细胞源性神经营养因子联合神经干细胞移植治疗脑室周围白质软化新生大鼠的疗效观察  

Transplant effect of neural stem cells combined with GDNF for the treatment of periventricular leukomalacia in neonatal rats

作　　者：贺月秋[1] 陈惠金[1] 钱龙华[1] 陈冠仪[1] 

机构地区：[1]上海交通大学医学院附属新华医院上海市儿科医学研究所,上海200092

出　　处：《临床儿科杂志》2009年第10期963-970,共8页Journal of Clinical Pediatrics

基　　金：上海市教委课题项目(No.05BZ08)

摘　　要：目的探讨在移植物中加入胶质细胞源性神经营养因子(glial cell line derived neurotrophic factor,GDNF),能否增强外源性神经干细胞(neural stem cells,NSCs)经脑室移植治疗脑室周围白质软化(periven-tricular leukomalacia,PVL)新生大鼠的移植效果。方法采用E14胎鼠大脑皮质制备NSCs。2日龄新生大鼠随机分为PVL对照组(PVL组),PVL+NSCs移植组(PVL+NSCs组),PVL+NSCs移植+GDNF组(PVL+NSCs+GNDF组),假手术对照组(Sham组),Sham+NSCs移植组(Sham+NSCs组),以及Sham+NSCs移植+GDNF组(Sham+NSCs+GDNF组)。对PVL新生大鼠在建模后72h进行立体定位仪下经脑室NSCs移植,分别于移植术后7、14、21d进行免疫荧光、光镜及电镜病理检测,评估在NSCs中加入GDNF对移植效果的可能影响。结果经脑室植入的外源性NSCs在脑内具有良好的迁移能力,3d内大部分移行至脑室周围,2周左右在脑室周围主要分化为少突胶质细胞前体,部份分化为神经元及星形胶质细胞。三种分化细胞在加入GDNF移植组显著多于未加GDNF移植组(P均<0.05)。移植后21d光镜下脑病理显示,两个移植组的脑白质病理均获明显改善,尤其加入GDNF移植组的脑白质重度病变发生率较未加GDNF移植组下降了25.5%(P<0.01)。移植后21d的电镜显示,PVL组罕见髓鞘形成,两个移植组的髓鞘形成明显增加,尤其加入GDNF移植组的髓鞘形成明显多于未加GDNF移植组。结论在外源性NSCs移植物内加入GDNF,可明显增强NSCs经脑室移植治疗PVL新生大鼠的移植疗效。Objective To explore whether the glial cell line derived neurotrophic factor （GDNF） can enhance the effect of neural stem cells （NSCs） implanted into ventricle for the treatment of periventricular leukomalacia （PVL） in neonatal rats. Methods NSCs were prepared from E14 embryonic rat brain. Two-day-old neonatal rats were randomly divided into the PVL group, the PVL ＋ NSCs group, the PVL ＋ NSCs ＋ GDNF group, the sham group, the sham ＋ NSCs group, and the sham ＋ NSCs ＋ GDNF group. The intraventricular transplantation of exdogenous NSCs with or without GDNF under stereo-positioner was undertaken in neonatal rats with either PVL or sham at 72 h after PVL model was established. The transplant effect of NSCs combined with GDNF was evaluated by immune fluorescence, light and electron microscopes at days 7, 14 and 21 after transplantation, respectively. Results The exdogenous NSCs implanted through ventricle had a favorable ability of migration. The most exdogenous NSCs migrated into the periventricular area within three days, and differentiated mainly into OLs precursors, and partly into neurons or astrocytes about two weeks after transplantation. The amount of three differentiated cells from NSCs in the NSCs with GDNF groups were more plenty than that in the NSCs without GDNF groups （P〈0.01 or 0.01 for all）. There was the significant improvement in cerebral white matter pathologically in both implanted group on day 21 after transplantation under light microscopy, in particular the percentage of serious abnormal white matter decreased 25.5 percentage point in the NSCs with GDNF groups compared to those in the NSCs without GDNF groups （P〈0.01 for all）. The obviously increased myelinogenesis in the white matter in both of PVL group with NSCs transplantation in particular in the NSCs with GDNF group, and sparse myelinogenesis in the PVL group without NSCs transplantation were observed under electron microcope on day 21 after transplantation. Conclusions It is indicated that GDNF can enhan

关 键 词：脑室周围白质软化 神经干细胞 细胞移植 胶质细胞源性神经营养因子 

分 类 号：R742.89[医药卫生—神经病学与精神病学]