目的:探讨不同的低氧浓度下离体培养胎鼠脑皮质神经干细胞(NSCs)的增殖分化及HIF-1α的表达情况,以此分析HIF-1α对神经干细胞的增殖分化作用.方法:体外低氧条件下(3%O2和10%O2)培养扩增胚鼠皮质来源的神经干细胞,实验组分为3%O2低氧1、3、5d组;10%O2低氧1、3、5d组,常氧为对照组.免疫细胞化学染色法鉴定神经干细胞及其子代细胞.RT-PCR检测实验组细胞HIF-1αmRNA的表达.结果:各低氧组NSCs内HIF-1αmRNA的表达量均明显高于对照组,且10% O2组与3% O2组相比,低氧5d HIF-1αmRNA 的表达量显著增加;实验组NSE阳性神经元的比率均明显高于对照组,10% O2组与3% O2组相比,低氧5d 10% O2组中NSE阳性神经元的比率明显增高.结论:低氧可促进NSCs增殖分化及HIF-1αmRNA的表达,且HIF-1αmRNA的表达量与低氧浓度及时间有关.%Objective:To observe the proliferation and differentiation of cortical neural stem cells (NSCs) from embryonic mice and the expression of hypoxia induciblc factor-la( HIF-la) under the different hypoxic concentrations in vitro, and to analyze the contribution of HIF-laon NSCs. Methods: NSCs from brain cortex of embryonic mice were cultured and ampiificatcd in lower O2 ( 3 % O2 and 10 % O2) in vitro , and all experimental dishes were divided into hypoxia for 1 , 3 and 5 days group ; The normoxic group was as control. Immunocytochcmical techniques were used to identify stem cells and their progenies , HIF-la mRNA expression in NSCs was examined by RT-PCR. Results : HIF-la mRNA was expressed significantly higher in every hypoxic group, and 10 % O2 group compared with 3 % O2 group, HIF-la mRNA expression increased significantly in hypoxia 5d . The ratio of NSE-positivc neurons in the experimental group were significantly higher than the control group , and 10 % O2 group compared with 3 % O2 group , the ratio of NSE-positivc neurons was significantly higher. Conclusion : Hypoxia can promote the proliferation and differentiation of NSCs and the expression of HIF-la mRNA , and the expression of HIF-la mRNA with hypoxia concentration and time-related.
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