酪氨酸羟化酶基因修饰的神经干细胞脑内移植PD模型动物旋转行为及神经生化研究

Study of PD model rats with intracerebral transplantation of TH-gene modified neural stem cells on their rotation and neurobiochemical changes

目的 探讨TH基因修饰的神经干细胞脑内移植对PD模型动物旋转行为和神经生化的影响。方法 构建pN2ATH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 ,测定移植后不同时间PD大鼠旋转行为改善以及DA和DOPAC含量变化。结果 TH基因修饰的神经干细胞移植 8周后能显著降低PD大鼠旋转行为 ,增加纹状体DA和DOPAC含量 ,疗效好于单纯神经干细胞移植组和对照组。结论 TH基因修饰的神经干细胞脑内移植能增加纹状体DA和DOPAC含量 ,降低PD大鼠旋转行为 ,从而对PD大鼠有明显的治疗作用 ,也为PD的治疗开辟了新途径。

Objective To study effects of intracerebral transplantation of TH gene modified neural stem cells (NSCs) on PD model rats' rotation and neurobiochemical changes. Methods The pN2ATH 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 N2ATH 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 apomorphine induced rotation and DA、DOPAC levels. Results The rotation of PD rats receiving grafts of TH gene modified NSCs decreased apparently 8 weeks after transplantation compared with the control graft group and NSCs group. The DA and DOPAC levels of the PD rats receiving TH transduced NSCs 8 weeks after transplantation were higher than that of receiving NSCs grafts and Saline grafts. Conclusion Intracerebral transplantation of TH gene modified NSCs can produce DA and DOPAC so that it can treat PD rats and may provide a new source of therapy for PD.