基因修饰的神经干细胞脑内移植后TH表达和神经递质的变化

TH-expression and neurotransmitter changes of PD rats with intracerebral transplantation of TH-gene modified neural stem cells

目的 探讨经TH基因修饰的神经干细胞脑内移植后 ,PD模型动物脑内TH表达和神经递质的变化。方法 构建 pN2 ATH逆转录病毒载体质粒 ,用PA3 17细胞包装 ,G418筛选阳性克隆 ,病毒上清感染神经干细胞 ,将神经干细胞和表达TH的神经干细胞植入PD大鼠纹状体内 ,观察移植后TH的表达情况以及DA和DOPAC含量变化。结果 TH基因修饰的神经干细胞移植 2月内 ,TH在纹状体内的表达增加 ,纹状体DA和DOPAC含量增高 ,好于单纯神经干细胞移植组和对照组 ,但比正常水平低。结论 TH基因修饰的神经干细胞 ,脑内移植能增加纹状体内TH的表达以及DA和DOPAC含量 ,为其脑内移植治疗PD提供了理论基础。

Objective To study TH-expression and neurotrasmitter changes of PD rats with intracerebral transplantation of TH-gene modified neural stem cells(NSCs).Methods The pN 2ATH recombinant retroviral plasmids were transferred into PA317 cells with liposome.Resistant colonies were cloned and maintained under G418 selection,and the virus supernatants were harvested.The NSCs were incubated in N 2ATH viral supernatant and selection under G418.NSCs and TH-gene modified NSCs were transplantation in the corpus striatum of the PD model rats.At different time after transplantation the rats were tested for TH expression,DA and DOPAC levels in striatal areas.Results TH expression,DA and DOPAC levels in striatal areas of the PD rats receiving TH-gene modified NSCs 2 months after transplantation were higher than that of receiving NSCs grafts and Saline grafts,but lower than that of normal rats.Conclusion Intracerebral transplantation of TH-gene modified NSCs can incerase TH expression,DA and DOPAC levels in striatal areas of the PD rats,which may provide its theoretic basis for PD therapy.