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目的:在体外研究Wnt/β-catenin信号对于少突胶质-GABA能神经元前体细胞分化的影响。方法:培养胎鼠中脑神经干细胞,在Wnt3a刺激或过表达β-catenin的条件下进行诱导分化,采用免疫细胞化学染色和Western Blot检测Wnt/β-catenin信号激活时,少突胶质-GABA能神经元共同前体细胞标记蛋白DLX2,少突胶质细胞标记蛋白NG2,GABA能神经元标记蛋白GAD67的表达。结果:Western Blot显示Wnt-3a处理上调β-catenin和Wnt/β-catenin信号下游靶基因cyclin D1和Axin2的表达;Wnt3a处理和过表达β-catenin的神经干细胞可显著上调Dlx2,NG2和GAD67阳性细胞的百分率(P<0.05)。Wnt3a刺激和β-catenin过表达均抑制神经干细胞向星型胶质细胞方向的分化。结论:Wnt/β-catenin信号可在体外促进神经干细胞向GABA能神经元和少突胶质细胞方向分化。
OBJECTIVE: To investigate the effects of Wnt / β-catenin signaling on differentiation of oligodendrocyte-GABAergic neurons in vitro. Methods: Neural stem cells of fetal rat midbrain were cultured and induced under Wnt3a stimulation or overexpression of β-catenin. Immunocytochemical staining and Western Blot were used to detect the activation of Wnt / β-catenin signaling in oligodendrocyte-GABA Can neuron common precursor cell marker protein DLX2, oligodendrocyte marker protein NG2, GABAergic neuron marker protein GAD67 expression. Results: Western Blot showed that Wnt-3a upregulated the expression of cyclin D1 and Axin2, a target gene downstream of β-catenin and Wnt / β-catenin signaling. Wnt3a-treated and overexpressed β-catenin-derived neural stem cells significantly upregulated the expression of Dlx2, NG2 and GAD67 The percentage of cells (P <0.05). Both Wnt3a stimulation and β-catenin overexpression inhibited the differentiation of neural stem cells to astrocytes. Conclusion: Wnt / β-catenin signaling can promote the differentiation of neural stem cells into GABAergic neurons and oligodendrocytes in vitro.