NGF/GDNF基因修饰神经干细胞移植对AD模型鼠前脑胆碱能神经元的保护作用

EFFECT OF TRANSPLANTATION OF THE NEURAL STEM CELLS MODIFIED WITH GENE OF NGF OR GDNF ON THE CHOLINERGIC NEURONS OF RASAL FOREBRAIN OF AD MODEL RAT

目的 观察NGF GDNF基因修饰神经干细胞 (NSC)单独和联合移植对穹窿海马伞 (FF)切断后胆碱能神经元的保护作用。 方法 将两种基因修饰的NSC单独和联合移植入FF切断的大鼠侧脑室。移植后 3周取脑行ChAT免疫组织化学染色 ,用Student NewmanKaels方法对内侧隔核 (MS)和斜角带垂直支 (VDB)的ChAT阳性细胞数 (损伤侧与正常侧的百分比 )进行计数分析。 结果 NGF组伤侧MS的ChAT阳性神经元数为 81% ,明显高于损伤组 (34% )、NSC组 (36 % )和GDNF组 (5 0 % ) ,P <0 0 1;NGF +GDNF组为 6 8% ,明显高于损伤组和NSC组 (P <0 0 1) ,亦高于GDNF组 (P <0 0 5 ) ;GDNF组与损伤组和NSC组相比 ,有统计学意义 (P <0 0 5 )。NGF组和NGF +GD NF组伤侧VDB的ChAT阳性神经元数分别为 80 %和 72 % ,明显高于损伤组 (5 6 % )和NSC组 (5 2 % ) ,P <0 0 1,亦高于GDNF组 (5 9% ) ,P <0 0 5 ;GDNF组与损害组和NSC组相比无明显差异 (P >0 0 5 )。 结论 NGF GDNF基因修饰NSC单独和联合移植均能不同程度地保护损伤的ChAT阳性神经元 ,NGF组和NGF +GDNF组作用较大 ,GDNF组作用较小。

Objective To investigate the effect of a single or combined transplantation of the neural stem cells(NSCs) modified with gene of NGF or GDNF on the cholinergic neurons of basal forebrain of AD model rat. Methods The NSCs modified with gene of NGF or GDNF were implanted in single or combined into the lateral cerebral ventricle of the rats after fibria\|fornix transection.The rats were killed three weeks after transplantation and the brain sections concluding basal forebrain were cut coronally on a freezing microtome and were processed by immunohistochemistry staining with antibodies against ChAT.The numbers of ChAT positive neurons of medial septum(MS) and vertical diagonal band(VDB) were analyzied statistically with one way of Student\|Newman Kaels. Results In MS,the percentages of ChAT positive neurons at the lesion side to the intact side in NGF group was 81% which was significantly higher than that in the lesion group(34%),NSC group(36%) and GDNF group(50%), P <0\^01 respectively.NGF+GDNF group was 68% which was also significantly higher than that in the lesion group and NSC group( P <0\^01) and GDNF group( P <0\^05).GDNF group was higher than that in the lesion and NSC groups( P <0\^05).In VDB,the percentages of ChAT positive neurons in NGF group and NGF+GDNF group were 80% and 72% which were significantly higher than those in the lesion group(56%) and NSC group(52%), P <0\^01 respectively, and GDNF group(59%), P <0\^05.But there was no significant difference among the GDNF group,lesion group and NSC group( P >0\^05). Conclusion\ The injury cholinergic neurons can be protected in different extent after a single or combined transplantation of the neural stem cells modified with gene of NGF or GDNF.Among these three groups,greater protection was found in NGF group and NGF+GDNF group,and lesser protection in GDNF group.\;[