GDNF对Nurr1基因的大鼠骨髓源神经干细胞的诱导分化作用

GDNF Induced Differentiation of SD Rat BMSCs-D-NSCs Transfected with Nurr 1 Gene

目的:利用胶质细胞系源性神经营养因子(glial cell line-derived neurotroph ic factor,GDNF)对经过转染核相关因子1(nuc lear related factor 1,Nurr1)基因的大鼠骨髓源神经干细胞(bone m arrow strom al cells de-rived from neural stem cells,BMSCs-D-NSCs)进行培养和诱导分化,研究其能否促进Nurr1-BMSCs-D-NSCs向多巴胺能神经元转化。方法:(1)构建AAV-pcDNA3.1-Nurr1载体;(2)诱导SD大鼠BMSCs分化为成熟的神经元样细胞;(3)用脂质体法转染Nurr1基因到大鼠BMSCs-D-NSCs后用GDNF进行培养和诱导分化。结果:(1)AAV-pcDNA3.1-Nurr1载体携带预期的Nurr1遗传信息;(2)Nurr1基因成功转染到BMSCs-D-NSCs中并且持续表达;(3)Nurr1-BMSCs-D-NSCs以GDNF培养后表达TH因子。结论:GDNF能促进经过转染Nurr1基因的大鼠BMSCs-D-NSCs向多巴胺能神经元定向分化。

Objective： To determine whether glial cell line-derived neurotrophic factor （GDNF） can induce differentiation of bone marrow stromal cells derived from neural stem cells （ BMSCs-D-NSCs ） transfected with Nurr 1 gene into dopaminergic neuron in SD rats. Methods： （1） A recombinant adeno-associated virus vector encoding Nurr 1 （ AAV-pcDNA3.1-Nurr 1 ） was constructed using techniques of molecular cloning; （2） The SD rat BMSCs were induced to differentiate into mature neuron-like cells; （3） The BMSCs-D-NSCs were cultured and induced with GDNF after the rats were transfected with Nurr 1 using Lipofectamin 2000. Results： （ 1 ） The AAV-pcDNA3.1-Nurr 1 vector contained desired genetic information encoding Nurr 1 ; （2） Nurr 1 was found successfully transfected and persistently expressed in BMSCs-D-NSCs; （3） The TH factor was expressed in BMSCs-D-NSCs after cultured with GDNF. Conclusion：GDNF can induce SD rat BMSCs-D-NSCs transfected with Nurr 1 gene differentiate into dopaminergic