核受体相关因子1基因转染骨髓源性神经干细胞体外诱导向多巴胺能神经元的分化

背景：核受体相关因子1基因修饰是否促进骨髓源性神经干细胞向多巴胺能神经元分化少见报道。 目的：观察核受体相关因子1基因修饰骨髓源性神经干细胞在体外诱导分化为多巴胺能神经元的作用。 方法：体外培养和纯化大鼠骨髓源性神经干细胞,将骨髓源性神经干细胞分为4组,将未转染的骨髓源性神经干细胞随机分为对照组,脑源性神经营养因子组;将筛选出的重组质粒转染阳性的神经干细胞分为核受体相关因子1组和核受体相关因子1+脑源性神经营养因子组。 结果与结论：RT-PCR显示转染的骨髓源性神经干细胞4 d后核受体相关因子1高表达,分化结果显示：核受体相关因子1+脑源性神经营养因子组细胞内酪氨酸羟化酶在mRNA水平上的表达量最高,神经干细胞贴壁分化后各组酪氨酸羟化酶阳性细胞比例均明显高于对照组,其中以核受体相关因子1+脑源性神经营养因子组分化比例最高,为(52.44±15.9)%。提示核受体相关因子1基因修饰可促进骨髓源性神经干细胞向多巴胺能神经元分化,并通过脑源性神经营养因子的诱导作用,在体外可获得大量的多巴胺能神经元。

Differentiation of bone marrow-derived neural stem cells transfected by nuclear receptor-related factor 1 gene into dopaminergic neurons in vitro

BACKGROUND:Few reports have addressed whether genetic engineering of bone marrow-derived neural stem cells transfected by nuclear receptor-related factor 1 (Nurr1) gene can enhance the differentiation into dopaminergic neurons. OBJECTIVE:To investigate the differentiation of bone marrow-derived neural stem cells transfected by Nurr1 gene into DA neurons in vitro. METHODS:Rat bone marrow mesenchymal stem cells were in vitro cultured and purified and then divided into four groups. The un-transfected bone marrow-derived mesenchymal stem cells were divided into control group and brain-derived neurotrophic factor (BDNF) group; the screened recombinant plasmid-positive neural stem cells were divided into Nurr1 group and Nurr1+BDNF group. RESULTS AND CONCLUSION:Plenty of Nurr1 in transfected bone marrow-derived mesenchymal stem cells were detected by RT-PCR after 4 days. Differentiation results showed that the mRNA expression of tyrosine hydroxylase in the cells of Nurr1+BDNF group was highest. The proportion of tyrosine hydroxylase positive cells in BDNF group, Nurr1 group and Nurr1+BDNF group was significantly higher than that in the control group after adherent differentiation of neural stem cells. The proportion of tyrosine hydroxylase-positive cells in the Nurr1+BDNF group was highest; the proportion was (52.44±15.9) %. The results indicate that the Nurr1 gene can promote bone marrow-derived mesenchymal stem cells to differentiate into dopaminergic neurons, and a large number of dopaminergic neurons are obtained after being induced by BDNF.