中文摘要：

目的利用慢病毒载体建立GFP-Nurrl基因修饰的原代神经干细胞（NSCs）模型并观察Nurrl过表达后NSCs向多巴胺神经元的分化影响。方法利用基因重组构建pLenO-DCE-Nurrl慢病毒载体，用慢病毒转染第三代NSCs，转染72h后荧光检测转染效果；设置空白对照组、空载体组及DCE-Nurrl组，分别用Westernblot及PCR检测Nurrl的表达差异；并将转染后的NSCs分化培养7d后分别用免疫细胞化学检测、Westernblot及PCR检测酪氨酸羟化酶（TH）的表达。结果慢病毒转染NSCs72h后，转染率可达90％，与对照组相比DCE-Nurrl组高表达Nurrl。经慢病毒载体感染后的NSCs仍具备分化潜能，分化培养后发现DCE-Nurrl组分化的神经细胞中TH阳性细胞分化率90．60％，对照组为21．2％。结论慢病毒载体可高效转染NSCs过表达Nurrl；Nurrl基因过表达可以促进中脑腹侧来源NSCs向TH阳性多巴胺能神经元方向分化。

英文摘要：

Objective To establish the GFP-Nurrl modified NSCs model via Lentiviral vectors and observe the im- pact on the differentiation of neural stem ceils by Overexpression of Nurrl. Methods pLenO DCE-Nurrl lentiviral vectors was constructed by genetic recombination technology. P3 NSCs were infected with lentiviral vectors and the effect of trans- fection was measured by fluorescence expression after 72 hours. The expression of Nurrl between three groups, which in- cludes control group, empty vector group and DCE-Nurrl group, was analyzed by Western blot/PCR respectively. The ex- pression of TH among three groups was analyzed via immunofluorescence, Western blot and PCR after 7-days differentiation culture of the transfected NSCs. Results Approximately 90% of NSCs can be transfected by pLenO-DCE-Nurrl after 72 hours. Nurrl expression was upregulated in DCE-Nurrl group, compared to control group. NSCs remains the capacity to dif- ferential after transfected by lentiviral vectors, and the TH positive cells was 90.60% , while only 21.2% in the control group. The expression of TH and DAT was also upregulated in DCE-Nurrl group. Conclusion NSCs overexpression of Nurrl can be mediated by lentiviral vectors with high efficiency. NSCs derived from embryonic mesencephalic can be in- duced to TH positive dopaminergic neurons via overexpression of Nurrl.