低氧条件下星形胶质细胞对神经干细胞增殖的影响  

Influence of astrocytes on proliferation of neural stem cells under the low oxygen condition

作　　者：崔晓萍[1] 陈建梅[2] 穆军山[1] 叶建新[1] 殷红兵[1] 林航[1] 

机构地区：[1]南京军区福州总医院神经内科,350025 [2]南京军区福州总医院骨一科,350025

出　　处：《临床神经病学杂志》2011年第6期448-451,共4页Journal of Clinical Neurology

基　　金：南京军区医学科技创新资助项目(08MA101);福建省青年人才基金资助项目(2008F3085)

摘　　要：目的探讨体外低氧条件下星形胶质细胞对海马神经干细胞(NSCs)增殖的影响。方法将分离出的新生小鼠海马NSCs分别与低氧条件星形胶质细胞培养液(低氧组)、低氧条件星形胶质细胞培养液+磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/Akt)通路拮抗剂LY294002(拮抗剂组)以及常氧条件星形胶质细胞培养液(对照组)共培养;分别于培养24 h、48 h、72 h应用四甲基偶氮唑盐(MTT)法检测各组NSCs活性;应用Western Blot法检测各组NSCs磷酸化Akt(p-Akt)和磷酸化糖原合成酶激酶(p-GSK)-3β含量。结果 (1)共培养24 h、48 h及72 h,低氧组NSCs活性的光密度(OD)值分别为0.292±0.006、0.382±0.005、0.649±0.028,拮抗组分别为0.197±0.003、0.255±0.005、0.325±0.012,对照组分别为0.107±0.006、0.198±0.008、0.254±0.006;低氧组各时间点NSCs活性显著高于拮抗剂组和对照组(均P<0.05);(2)共培养24 h、48 h和72 h,低氧组NSCs p-Akt和p-GSK-3β的吸光度(A)值分别为0.52、0.71、0.86及0.49、0.65、0.82;拮抗剂组分别为0.39、0.42、0.61及0.31、0.35、0.40;对照组分别为0.34、0.38、0.39及0.28、0.31、0.35;低氧组各时间点p-Akt及p-GSK-3β含量显著高于拮抗剂组和对照组(均P<0.05)。结论缺氧刺激可激活星形胶质细胞PI3K/Akt信号通路,促进NSCs增殖。Objective To investigate the influence of astrocytes on proliferation of hippocampal neural stem ceils （NSCs） under the low oxygen condition in vitro. Methods The obtained hippocampal NSCs of neonate mice were cocultured under the low oxygen condition astrocyte cuhures （low oxygen group）, low oxygen condition astrocyte cultures ＋ PI3K/Akt path antagonists LY294002 （antagonists group） and normal oxygen condition astrocyte cultures （control group）. After 24 h, 48 h, 72 h, the NSCs activity was detected by four methyl accidentally nitrogen salt （MTT） method; the contents of phosphorylated Akt （p-Akt） and phosphorylated Glycogen synthase kinase （p- GSK） -36 in NSCs were detected by Western Blot method. Results （1） Cultured 24 h,48 h,72 h, the optical density （OD） value of NSCs activity was 0. 292 ± 0. 006, 0. 382 ± 0. 005, 0. 649 ± 0. 028 rseparately in low oxygen group, which was 0. 197 ±0. 003, 0.255±0. 005, 0. 325 ±0. 012 rseparately in antagonists group, 0. 107±0. 006, 0. 198 ±0.008, 0. 254±0. 006 rseparately in control group. The NSCs activity in low oxygen group at each time point was significant highr than antagonists and control groups （ all P 〈 0. 05 ）. （ 2 ） Cultured 24 h, 48 h, 72 h, the absorbance（A） value of p-Akt and p-GSK-3β content in NSCs were 0.52, 0.71, 0.86 and 0. 49, 0.65, 01 82 rseparately in low oxygen group, which was 0.39, 0. 42, 0. 61 and 0.31, 0. 35, 0.40 rseparately in antagonists group, 0.34, 0.38, 0.39 and 0.31, 0.35, 0.40 rseparately in eontral group. The contents of p-Akt and p-GSK-313 in NSCs in low oxygen group at each time point were significant higher than antagonists and control groups （ all P 〈 0. 05）. Conclusion Hypoxia stimulation can activite the PI3K/Akt signal path of astrocyte and promote NSCs proliferation.

关 键 词：低氧 神经干细胞 星形胶质细胞条件培养液 增殖 磷脂酰肌醇-3激酶/蛋白激酶B 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]