嗅鞘细胞移植促进脑损伤大鼠神经功能恢复及其作用机制研究

目的 观察嗅鞘细胞移植对脑损伤大鼠神经功能恢复及神经元存活的影响并探讨其作用机制.方法 对经培养纯化的嗅鞘细胞作NGFRp75免疫细胞化学染色鉴定并制备移植用细胞悬液,部分细胞预作荧光标记用于移植后观察存活情况;24只SD大鼠平均分成3组:假手术无损伤组(A组),大鼠脑皮质运动区损伤组(B组),相同脑损伤后嗅鞘细胞移植组(C组);术后当天及第3,7,14天分别对各组动物进行神经功能缺损严重程度(NSS)评分;术后第14天取损伤部位组织作神经元核心抗原免疫组化(NeuN)检测并计数阳性细胞数量;数据采用SPSS17.0软件进行统计学分析.结果 (1)培养的嗅鞘细胞呈NGFRp75阳性,阳性率90%;(2)移植14d后核荧光标记的嗅鞘细胞在宿主体内存活良好;(3)术后第14天B组NSS评分为[(2.00±0.53)分],C组为[(1.25±0.46)分],明显优于B组(P<0.05);(4)NeuN阳性细胞计数B组为[(39.2±7.1)分],C组为[(45.8±6.0)分],明显优于B组(P<0.05).结论 嗅鞘细胞移植可明显促进脑损伤大鼠神经功能恢复,其机制可能与嗅鞘细胞促进宿主神经元存活有关.

The olfactory ensheathing cells transplantation for traumatic brain injury recovery of nerve function and its mechanism

Objective To observe olfactory ensheathing cells transplantation on brain injury recovery nerve function and to explore its mechanism. Methods After purification of the olfactory ensheathing cells cultured for NGFRp75 immunocytochemical identification and preparation of cell suspension for transplantation, some cells were pre-labeled for bservation of survival after transplantation. 48 adult SD rats were randomly divided into three groups:sham operation without injury group (A group),cerebral cortex motor area injury group (B group) , the same brain injury and the olfactory ensheathing cell transplantation group (C group). At postoperative day,3 d, 7 d,14 d the neurological severity score (NSS) of rats were assessed; 14 d after injury of brain tissues were taken for NeuN immunohistochemistry host the number of neurons change. The data were statistically analyzed using SPSS17.0 software. Results (1) Cultured olfactory ensheathing cells showed NCFRp75 positive,the positive rate was 90%. (2) 14 d after transplantation of nuclear fluorescence labeling of olfactory ensheathing cells survived well in the host body. (3) 14 d after NSS score of B group( 2.00 ± 0.53) and C group ( 1.25 ± 0.46) were significantly better than the B group (P<0.05). (4) NeuN positive cells in B group (39.2 ±7. 1) and C group(45, 8 ± 6.0) were significantly better than B group (P<0.05). Conclusions Olfactory ensheathing cell transplantation can promote the recovery of neurological function in rats brain injury,which may be related with olfactory ensheathing cells to promote neuron survival in the host.