稳定表达GDNF基因的骨髓基质干细胞对多巴胺能神经元的作用

目的 制备胶质细胞源性神经生长因子(GDNF)基因修饰的骨髓基质干细胞(MSCs)，观察其对多巴胺能神经元的作用，探索治疗帕金森氏病的新途径。方法 应用逆转录聚合酶链反应(RT-PCR)方法从新生小鼠大脑皮层细胞克隆出GDNF cDNA片断，以pEGFP-C1为载体导入MSCs，制备稳定表达GDNF基因的MSCs工程细胞，采用联合培养的技术通过倒置显微镜和免疫组织化学的方法观察MSCs和GDNF基因修饰的MSCs工程细胞与多巴胺能神经元之间的相互作用。结果 MSCs和GDNF基因修饰的MSCs工程细胞均能促进多巴胺能神经元的存活和生长，MSCs工程细胞作用更强。结论 成功构建了GDNF基因修饰的MSCs工程细胞，该细胞对多巴胺能神经元有明显营养保护作用，在帕金森病治疗中可能有重要价值。

Neurotrophic and protective effection of GDNF gene modified MSCs on primary dopaminergic neurons of mice

Objective To construct a kind of engineered cell secreting GDNF,observe its protective effect on primary dopaminergic neurons and study its possible effects on gene therapy of Parkinson's disease.Methods Mouse GDNF cDNA was amplified from newly-born mouse cortex cell of cerebrumby RT-PCR,and ligated into the pEGFP-C1 vector.Then transferring the recombined plasmid into MSCs and screening engineered MSCs which can secrete GDNF protein stably.Dopaminergic neuron cells were co-cultured with MSCs and MSCs which modified by GDNF gene respectively,the effects between these cells were observed using phase-contrast microscope and immunohistochemistry staining.Results Both MSCs and GDNF-modified MSCs showed great neurotrophic and protective effections on dopaminergic neurons,but the effections of GDNF-modified MSCs were stronger than MSCs.Conlusions The GDNF-modified MSCs have been constructed successfully.These cells obviously promoted the survival and growth of dopaminergic neurons.