《中国康复理论与实践》

《中国康复理论与实践》 ›› 2007, Vol. 13 ›› Issue (09): 850-852.

• 基础研究 • 上一篇    下一篇

氯化锂对培养神经干细胞增殖分化的影响及其机制研究

景秀京1; 杨继飞1; 张建2; 胡远兵1; 杨忠2   

  1. 1.四川成都武警总队医院,四川成都市 610041;2.第三军医大学神经生物学教研室 重庆市 400038
  • 收稿日期:2006-12-26 出版日期:2007-09-01 发布日期:2007-09-01

Effect of Lithium Chloride on Proliferation and Differentiation of Neural Stem Cells

JINGXiu-jing, YANG Ji-fei, ZHANGJian,et al   

  1. Chengdu Hospital of the Chinese People's Armed Forces, Chengdu 610041, Sichuan, China
  • Received:2006-12-26 Published:2007-09-01 Online:2007-09-01

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被引次数

2

摘要： 目的观察氯化锂对培养神经干细胞(NSCs)增殖及分化的影响,探讨Wnt信号途径对NSCs增殖分化的调控作用。方法分离培养并传代扩增NSCs,显微观察结合流式细胞仪分析不同浓度氯化锂处理对NSCs形态、贴壁分化及细胞周期动力学的影响,Western印迹定量检测不同浓度氯化锂作用后NSCs糖原合酶激酶3β(GSK3β)、β-catenin的表达变化。结果氯化锂作用后NSCs更多地保持悬浮生长的状态,离散细胞密度显著增加,神经球有减小分散趋势;氯化锂能显著抑制血清诱导的NSCs分化,表现为贴壁时间延长,细胞突起较对照组明显减少、缩短;流式细胞仪分析显示,氯化锂作用组NSCs处于S期的比例明显增高,G1期细胞呈现减少的趋势。且上述效应呈现一定的剂量效应关系。Western印迹显示,氯化锂能逐步增强β-catenin的表达,高浓度组能明显抑制GSK3β表达。结论氯化锂能明显抑制NSCs分化,刺激其增殖,该效应可能与其对Wnt信号途径的活化有关。

关键词: 氯化锂, 神经干细胞(NSCs), 糖原合酶激酶3β, 增殖, 分化

Abstract: Objective To investigate the effect of lithium chloride(LiCl),an inhibitor of glycogen synthase kinase-3beta(GSK-3beta),on proliferation and differentiation of neural stem cells(NSCs).Methods The NSCs were isolated from cortex of rat fetus and expanded in culturing system.Their morphological changes and attachment process were observed under microscope.The cell cycle dynamics of NSCs was examined with flow cytometry.And the expression of GSK-3β and β-catenin was examined quantitatively with Western blot.Results The culturing NSCs treated with LiCl were usually floated and much dispersed in the media.Many of the neurospheres became small and the time of attachment after serum induction became longer.Using flow cytometry,it was detected that the proportion of G1 phase NSCs declined gradually accompanying the increased concentration of LiCl,while the percentage of S and G2/M phase cells showed an increasing trend.Western blotting results revealed β-catenin expression increased whereas Gsk-3βdecreased gradually under the treatment of LiCl and also showed a dose dependent manner.Conclusion These results suggest that LiCl may promote the proliferation of NSCs and prevent them from differentiating,which may partly involve the activation of wnt/β-catenin signaling pathway.

Key words: lithium chloride, neural stem cells, glycogen synthase kinase-3beta, proliferation, differentiation