Ndrg2对培养大鼠神经干细胞增殖和分化的影响

Effects of Ndrg2 on the proliferation and differentiation of cultured rat neural stem cells

目的:检测Ndrg2在培养大鼠神经干细胞(NSCs)中的表达情况,探索改变Ndrg2表达对NSCs增殖和分化的影响。方法:用免疫细胞化学染色和Western Blot法检测Ndrg2在培养NSCs中的表达;利用AAV-Ndrg2过表达病毒及LV-Ndrg2-RNAi干扰病毒感染大鼠NSCs后,通过BrdU掺入实验观察干预Ndrg2表达后NSCs增殖的变化,利用神经元标记物Tuj1和星型胶质细胞标记物GFAP的免疫细胞化学染色分析NSCs的分化情况。结果:Ndrg2高表达于大鼠NSCs中;与对照组(感染病毒空载体)相比,上调Ndrg2表达可显著增加BrdU^+和Tuj1^+细胞数(P<0.05),但GFAP^+细胞数目无明显变化(P>0.05);相反,下调Ndrg2表达可减少BrdU^+细胞数(P<0.05),但不影响Tuj1^+和GFAP^+细胞数(P>0.05)。结论:Ndrg2表达于大鼠NSCs,它可正性调控NSCs的增殖并促进NSCs向神经元分化。

Objective：To study the expression of Ndrg2 in cultured rat neural stem cells （NSCs） and to explore its effects on the proliferation and differentiation of NSCs.Methods：Immunofluorescence staining and Western Blot were used to examine the expression of Ndrg2 in cultured rat NSCs;After over-expression of Ndrg2 and knockdown expression of Ndrg2 in NSCs transfected by AAV-Ndrg2 virus and LV-Ndrg2-RNAi virus,respectively,BrdU pulsing assay was used to examine NSC proliferation after perturbation of Ndrg2 expression,and immunofluorescence staining of neuronal maker Tuj1 and astrocytic maker GFAP were used to observe NSC differentiation.Results：Ndrg2 was highly expressed in the cultured rat NSCs.Compared with the control group （infected by vehicle virus）,Ndrg2 over-expression significantly increased the number of BrdU ＋ and Tuj 1 ＋ cells （P 〈 0.05）,but did not change that of GFAP ＋ cells （P 〈 0.05）.By contrast,Ndrg2 knockdown decreased the number of BrdU＋ cells （P 〈0.05）,but did not affect the number of Tuj1 ＋ and GFAP ＋ cells （P 〈 0.05）.Conclusion：Ndrg2 was expressed in rat NSCs,and may regulate NSC proliferation positively and promote NSCs differentiation into neurons.