神经干细胞移植配合强制性使用运动疗法对猕猴脑损伤功能恢复的影响及机制研究

Both constraint-induced movement therapy(CIMT) and neural stem cells transplantation therapy could significantly improve the neurological function recovery which may cause by brain injury. But until now, the mechanism of these therapies are still unclear. It may be related with nerve regeneration and angiogenesis. Due to the comparability anatomy & function of nerve system between human and rhesus monkey (Macaca mulatta), this project programs to study the mechanism of CIMT and neural stem cells transplantation on improvement of neurological function using rhesus monkey，which it is not reported up-to-date. The motor cortex is located by an electrophysiological stimulator. The rhesus monkey model of motor cortex injury is made by pumped. NF, MAP2, GAP-43, GFAP, VEGF, ANG and bFGF will be detected nearby the traumatic brain injury area. The survival and migration of transplanted neural stem cells will be also examined. The diameters and branches of the microvasculature among the damaged area will analysis combined with the confocal microscope which marked by the fluorescence. The cerebral blood flow changes of the injury related brain regions will determinate by SPECT. The neurological function before and after CIMT and the neural stem cells transplantation therapy will be evaluated by the scales. All of the results may reveal the possible mechanism of CIMT and neural stem cells transplantation therapy.

强制性使用运动疗法（CIMT）和神经干细胞移植均可明显改善脑损伤患者神经功能，机制尚不清楚，可能与促进损伤周围区神经再生或血管新生有关。本项目拟选猕猴为实验对象，用神经电生理定位捣毁大脑中央前回运动区制成脑损伤模型，运用神经干细胞移植配合CIMT训练动物，此项研究国内外未见报道。通过免疫组化法检测NF、MAP2、GAP-43、GFAP、VEGF、ANG、bFGF在脑损伤周围区神经元和胶质细胞表达变化；观察神经干细胞存活及迁移情况；荧光素标记微血管，结合共聚焦显微镜分析损伤区微血管的直径、面积及血管分支的变化；SPECT 测定损伤相关脑区脑血流量改变；神经量表评定训练前后动物神经功能变化；初步研究神经干细胞移植配合CIMT训练促进神经再生、血管新生机制，为临床开展CIMT治疗技术提供理论依据。

由国家自然科学基金委资助的项目“神经干细胞移植配合强制性使用运动疗法对猕猴脑损伤功能恢复的影响及机制研究”（计划资助时间：2013.01-2016.12，编号：81260298），本项目自2013年1月起按照计划任务书要求一直开展着相应的研究，由于实验动物因素，经申请国家自然科学基金委同意，改变了研究方向，主要侧重于移植干细胞是怎样发挥功能作用的基础研究，为干细胞替代治疗提供临床基础依据；在此基础上，进一步研究神经干细胞移植和强制性使用运动疗法促进脑损伤恢复的机制，为开展CIMT技术提供理论依据。我们已经完成了大鼠海马损毁后干细胞移植、猕猴下丘损毁及干细胞移植后的功能整合的研究和PD猕猴模型的建立，有关强制性使用运动疗法技术对PD猕猴影响的研究正在进行中。本研究首次在清醒动物上证实移植神经干细胞分化的神经元能够功能性整合到宿主神经网络中，为干细胞临床替代疗法提供了坚实的基础理论数据。截至2016年12月，现已经基本完成实验的主要部分，后续的实验及数据整理正在进行中，故向国家自然科学基金委申请结题。