低糖低氧对神经干细胞增殖和代谢的影响  

Effect of low glucose and/or hypoxia on the proliferation and metabolism of neural stem cells

作　　者：赵彤[1] 黄欣[1] 朱玲玲[1] 熊雷[1] 张宽[1] 吴丽颖[1] 刘兵[2] 吴奎武[1] 范明[1] 

机构地区：[1]军事医学科学院基础医学研究所脑保护与可塑性研究室,北京100850 [2]军事医学科学院307医院,北京100850

出　　处：《中国应用生理学杂志》2010年第4期412-415,共4页Chinese Journal of Applied Physiology

基　　金：973资助项目(2006CB504100);国家自然科学基金资助项目(30670792)

摘　　要：目的:本研究旨在探讨低糖低氧对大鼠神经干细胞增殖和代谢的影响。方法:实验采用不同葡萄糖浓度的培养基以及不同的氧浓度进行处理:高糖(4.5g/L)、低糖(1.4g/L);常氧(20%O2)、低氧(3%O2);神经干细胞(NSCs)来自孕13.5d的大鼠中脑,在不同糖浓度下培养至第三代进行低氧处理,分为低糖常氧(L+N)、低糖低氧(L+H)、高糖常氧(H+N)、高糖低氧(H+H)组。神经干细胞在上述四种条件下分别培养1、3、5d后,利用CCK-8检测神经干细胞的增殖情况;生化分析仪测定细胞培养上清液中葡萄糖、乳酸、丙酮酸浓度;RT-PCR方法检测葡萄糖转运蛋白4(GluT4)、葡萄糖激酶(GK)、丙酮酸激酶(PK)和乳酸脱氢酶(LDH)的表达变化。结果:在低糖低氧条件下培养3d时NSCs的数量增加最为明显;低糖低氧条件下,葡萄糖浓度降低最为显著;而丙酮酸浓度在低糖处理组均高于高糖处理组;同样地,在低糖低氧处理组培养上清中乳酸含量增加的幅度最大;此外,在低糖或低氧时Glut4和PK的表达也明显高于对照组。结论:低氧能促进NSCs的增殖,而以低氧和低糖共同作用时更为明显;在低氧低糖条件下,神经干细胞的代谢发生变化,葡萄糖的利用明显增加,主要通过糖酵解途径代谢产能。Objective：Recent study demonstrated that hypoxia could regulate the proliferation and differentiation of neural stem cells in vitro. In the present study,effects of low glucose and/or hypoxia on the proliferation and metabolism of neural stem cells were investigated in vitro. Methods： The neural stem cells were isolated from the rat embryonic mesencephalon （E13.5）,and exposed to different oxygen concentrations （low oxygen： 3% O2 or normaxia： 20% O2） and different glucose concentrations （high glucose concentration： 4.5 g/L and low glucose concentration： 1.4 g/L）for 3 days. The proliferation of neural stem cells were examined by CCK-8 assay. Furthermore,the content of glucose、lactate、and pyruvic acid in the medium were measured after cultured in different condition for 1,3,5 days. Results： Low oxygen and low glucose could increase the proliferation of neural stem cells respectively; in addition,the number of neurospheres under both low oxygen and glucose was the most among the four groups. The content of glucose and pyruvic acid in the medium from low oxygen or low glucose condition decreased,while the lactate concentration increased compared with the control group. Conclusion： The results indicate the neural stem cells prefer grow under the low glucose and low oxygen condition,and that is mainly under going glycolysis to maintain its self-renew ability. This study may provide us a useful clue for application of neural stem cells transplantation.

关 键 词：神经干细胞 低氧 低糖 代谢 

分 类 号：Q251[生物学—细胞生物学]