目的探讨利用转基因神经干细胞移植重建臂丛神经根性撕脱伤后运动功能的可行性。方法以PGET-1 TA质粒为克隆载体体外扩增NT-3基因,以含有增强型绿色荧光蛋白基因的pLEGFP-C1质粒为表达载体将体外扩增的NT-3基因导入神经干细胞内,并将该转基因神经干细胞移植于臂丛神经根性撕脱伤动物模型的C6脊髓节段内,观察移植细胞的存活、分化及迁移等。观察实验动物的运动功能变化,利用神经电生理检测及辣根过氧化物酶逆行神经示踪技术评价周围神经再生情况。结果NT-3基因成功导入神经干细胞内,该转基因神经干细胞移植于臂丛神经根性撕脱伤动物模型的C6脊髓节段内能够生存,并分化为神经元。实验动物的运动功能有所改善,但肌力低于3级。神经电生理检测及辣根过氧化物酶逆行神经示踪技术评价显示转基因神经干细胞移植能够促进周围神经再生。结论利用转基因神经干细胞移植治疗臂丛神经根性撕脱伤是一种相对有效的实验性治疗方法,但尚需进一步研究以提高疗效。
Objective To explore a new method utilizing transplantation of neural stem cells (NSC) modified by neurotrophin-3 (NT-3) gene to treat brachial plexus root avulsion experimentally. Methods The NT-3 gene was transferred into NSCs via the plasmid vectors. The untransfected NSCs,the pLEGFP-C1 treated NSCs, and the pLEGFP-C1-NT-3 treated NSCs were transplanted into spinal cord of animals with brachial plexus root avulsion. The survival,differetiation and migration of transplanted cells were determined under confocal laser scanning microscopy or by immunohistochemistry method. The neural regeneration was evaluated by gross observation, electrophysiological examination and reverse HRP tracing. Results The NT-3 gene was successfully amplified and transferred into NSCs via the plasmid vectors. The transplanted cells survived, differentiated and migrated and NT-3 was expressed within spinal cord. The animals regained some certain muscle strength which was less than 3 degree. The results of dectrophysiological examination and reverse HRP tracing in the pLEGFP-C1-NT-3 group were superior to the NSCs untransfected group or the pLEGFP-C1 group. Conclusion Transplantation of NSCs modified by NT-3 gene is a relatively effective way to treat brachial plexus root avulsion experimentally.