缺氧培养对大鼠神经干细胞体外增殖影响及其机制的研究

目的：研究应用缺氧对体外培养的大鼠神经干细胞增殖的影响。方法：将细胞分为4小时缺氧组、12小时缺氧组、促红细胞生成素（EPO）中和抗体组、IgG组以及对照组．测定大鼠神经干细胞经缺氧培养后的各组细胞克隆形成率以及EPO的表达变化。结果：单细胞培养条件下，与对照组相比，4小时缺氧组和IgG组克隆形成率明显增高；中和抗体组无明显变化；12小时组克隆形成率降低。但无统计学意义。缺氧4小时后，EPO蛋白在预处理后即刻出现表迭，4h达高峰，8h仍有部分表达。结论：短时间缺氧可以促进神经干细胞增殖．长时间缺氧则作用相反。缺氧对NSCs增殖作用的影响可能是由EPO介导产生。

Effect of hypoxia on proliferation protection and EPO expression of rats' neural stem cells in vitro

Objective: To investigate the effect of hypoxia on proliferation of rats' neural stem cells(NSCs) in vitro . Methods: A rat NSCs culture model was established. The NSCs were divided into 5 groups: namely, 4 hours group, 12 hours group, EPO(= erythropoietin) antibody group, IgG group and control group. Cloning efficiency of each group was determined after applying hypoxia. EPO expression was studied by western blotting. Results In mono-eell test, compared with control group,cloning efficiency of cells in 4 hours group and IgG group was obviously higher than that in control group. There was no significant difference between that in EPO antibody group and control group. Cloning efficiency of cells in 12 hours group was lower than that in control group with no statistical difference. Western blot analysis demonstrated a significant increase in Epo expression in NSCs cultures immediately after a 4 hours hypoxia. The increase peaked at 4 hours after preconditioning and was sustained till 8 hours. Conclusion: Hypoxia of 4 hours could enhanced the cell proliferation, while 12 hours hypoxia inhibited cell proliferation. EPO may play an important role in this process.