神经干细胞侧脑室内移植治疗脑瘫鼠的实验研究

Treatment of cerebral palsy by transplantation of neural stem cells into the lateral ventricles in rats

目的探讨神经干细胞(NSCs)侧脑室内移植对脑瘫鼠的治疗效果。方法将60只7日龄Wistar大鼠随机分为正常组、脑瘫组、NSCs移植组和神经营养因子(NT-3)修饰的NSCs移植组。除正常组外其余各组鼠均行左侧颈动脉结扎并缺氧2h建立脑瘫模型,建模3d后分别对两移植组进行侧脑室NSCs以及NT-3修饰的NSCs移植。移植后第46天,开始对各组大鼠的学习记忆以及行为学能力进行评估,学习记忆能力连续测5d。之后取鼠脑组织行病理学检测。结果①Morris水迷宫:正常组、两组移植组大鼠逃避潜伏期与脑瘫组比较明显缩短(P<0.05);②足印试验:正常组、两移植组潜伏期与脑瘫组比较明显缩短(P<0.05),右侧后肢足印重复也明显好于脑瘫组,足印间距减小,且稳定(P<0.05);③平衡木试验:与脑瘫组比较,正常组、移植组潜伏期时间明显缩短,后足滑脱次数明显减少(P<0.05);④组织病理学HE染色结果:脑瘫组鼠侧脑室周围细胞肿胀、变性、坏死和排列紊乱,局部囊性变和炎性细胞聚积;NSCs移植组鼠脑组织结构破坏以及细胞形态较脑瘫组有所改善;NT-3移植组恢复情况更好,细胞形态趋于正常。结论经侧脑室移植外源性NSCs可明显改善脑瘫鼠脑组织的病理损伤和促进其神经行为学及学习记忆功能的恢复。

Objective To study the curative effect of transplantation of neural stem cells （NSCs） into the lateral ventricles on cerebral palsy in rats. Methods Wistar rats （n=60） were randomly divided into normal（A）, model（B）, NSCs transplantation（C） and neurotrophin-3 （NT-3）-modified NSCs transplantation（D） groups of 15 animals each. The cerebral palsy model was made by the ligation of the left carotid artery followed by exposure to hypoxia （8% 02 ± 92% N2） for 2 hours. NSCs and NT-3-modified NSCs were transplanted lateral ventricles in groups C and D 3 days after the establishment of the model. The spatial memory and ethological function were determined and then the pathological examination of cerebral tissues was performed in all the groups. Results ①Morris water maze experiment showed that escape latent period was significantly shorter in groups A, C and D than that in group B （P〈0.05）. （② Footprints gait repeat spacing analysis test showed that the latent period was significantly shorter and the right footprints spacing was significantly smaller and stabler in groups A, C and D than those in group B （P〈0,05）. ③Balanee beam test showed that the time through the balance beam was significantly shorter and the times of rats＇ right leg slip from the beam is significantly less in groups A, C and D than those in group B （P〈O.05）. （~Pathological observation showed that the cells swelling, degeneration, necrosis, arrangment disorder, local cystic change and inflammatory cells accumulation were found around the lateral ventricles in group B, and the change in the cell morphology was slighter in groups C, especially in group D than that in group B. Conclusion The morphology and function of injured cerebral tissues can be significantly improved by the transplantation of exogenous NSCs or NT-3-modified NSCs into the lateral ventricles in the rats with experimental cerebral palsy.