目的 研究经侧脑室内注入人脐血间充质干细胞(hUCB-MSCs)联合鼠神经生长因子(mNGF)对脑瘫大鼠治疗效果。方法 采集足月新生儿脐带血100 ml,分离、培养、纯化人脐血间充质干细胞并进行检测和鉴定。将七日龄SD大鼠100只随机分为正常组10只、实验组(缺血缺氧性脑瘫模型)90只,实验组随机分为5组脑瘫模型组(CP组)、磷酸盐缓冲溶液(PBS)移植组、mNGF移植组、hUCB-MSCs移植组及hUCB-MSCs联合mNGF移植组。造模成功后1周,将hUCB-MSCs、mNGF分别注入左侧侧脑室,假移植组注射PBS液,移植后第1周、2周、3周,观察5溴-2脱氧尿核苷(BrdU)标记的脐血MSCs的存活、迁移,移植后第4周,对每组大鼠的神经行为学能力进行评估并检测大鼠脑组织胶质纤维酸性蛋白抗体(GFAP)和神经元特异性烯醇化酶(NSE)的表达及观察脑组织的形态学变化。结果 1)人脐血MSCs主要分布在左侧侧脑室、额叶、顶叶皮质及白质,随时间进展,向四周脑组织转移,Brdu、NSE、GFAP阳性细胞逐渐减少,差异有统计学意义(P<0.05);2)行为学评价与模型组及假移植组比较,单移植组、联合移植组的足印潜伏期、迷宫逃避潜伏期缩短,足印重复间距及后足滑脱次数减小,联合移植组优于单移植组,差异有统计学意义(P<0.05);3)HE染色MSCs和mNGF单移植组大鼠细胞形态及脑组织结构破坏较脑瘫组有所改善;联合移植组恢复情况更好,细胞形态恢复较单移植组好。结论 外源性脐血MSCs联合mNGF经侧脑室移植,可改善脑瘫大鼠的远期行为及脑损害。

Objective To study the treatment effect of human umbilical cord blood mesenchymal stem cells(hUCBMSCs) combined with mouse nerve growth factor (mNGF) to the cerebral palsy rats through lateral intraventricular transplantation. Methods 100 ml cord blood from full-term newborn was collected and to separate,culture and purify hUCBMSCs then detect and identify them.100 seven-day-old SD rats were randomly divided into 10 normal group and 90 experiment group(cerebral palsy produced by ischemia and hypoxia ) while the experiment group were split into cerebral palsy (CP) model group,phosphate buffer solution (PBS) transplantation group,mNGF transplantation group,human umbilical cord blood-derived mesenchymal stem cells transplantation group,and human umbilical cord blood-derived mesenchymal stem cells combined with mouse nerve growth factor transplantation group.After 1 w modeling success MSCs was injected,mNGF and PBS into Lateral ventricles.After transplantation in 1 w,2 w and 3 w,the survival and migration of MSCs with 5 bromine -2 (BrdU) labeled were observed.In the fourth week,the neural behavior ability of rats were studied and the rat brain tissues were taken to detect the expression of its GFAP and NSE. Results 1)The survival,migration of hUCBMSCs mainly distributed at the site of the left lateral ventricle,frontal lobe,parietal cortex and white matter.Along with the time going,its migration was to the surrounding brain tissue.BrdU,NSE,GFAP positive cells decreased gradually,and the difference was statistically significant (P<0.05).Compared with model group and sham transplantation,the footprints latency and maze escape latency of single transplantation groups and combined transplantation group were shorten,footprints repeat spacing and foot slippage number decreasing as well,and transplantation group was better than that of single transplantation groups,the difference was significant (P<0.05);2)HE staining:the cell morphology and brain tissue damage in the single transplantation group were better than that of the cerebral palsy group,and the recovery of the combined transplantation group was better than that of the single tran-splantation group. Conclusion Through intraventricular transplantation,exogenous MSCs transplantation combined with mNGF can improve the long-term behavior and brain damage in cerebral palsy rats.