解剖学报

解剖学报 ›› 2016, Vol. 47 ›› Issue (5): 577-582.doi: 10.16098/j.issn.0529-1356.2016.05.001

• 神经生物学 •    下一篇

自然衰老小鼠室管膜下区神经干细胞增龄性改变

董传明 金国华*   

  1. 南通大学医学院人体解剖学系，江苏省神经再生重点实验室，江苏 南通 226001
  • 收稿日期:2016-05-23 修回日期:2016-06-07 出版日期:2016-10-06 发布日期:2016-10-06
  • 通讯作者: 金国华 E-mail:jguohua@ntu.edu.cn
  • 基金资助:

    国家自然科学基金青年基金;江苏省高校自然科学基金面上项目;南通市科技计划项目

Age-related changes of neural stem cells from the subventricular zone of aged mice

DONG Chuan-ming JIN Guo-hua*   

  1. Department of Anatomy，Jiangsu Key Laboratory of Neuroregeneration，School of Medicine，Nantong University，Jiangsu Nantong 226001，China
  • Received:2016-05-23 Revised:2016-06-07 Online:2016-10-06 Published:2016-10-06
  • Contact: JIN Guo-hua E-mail:jguohua@ntu.edu.cn

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摘要：

目的 探讨自然衰老小鼠室管膜下区（SVZ）神经干细胞（NSCs）在衰老过程中生物学特点的改变。 方法 2月龄、28月龄小鼠，各10只，体外分离、培养、鉴定NSCs，5-溴-2-脱氧尿苷（BrdU）染色比较两组细胞的增殖水平；β-半乳糖苷酶（SA-β-Gal）染色检测细胞衰老水平；细胞内活性氧（ROS）试剂盒检测细胞ROS表达水平；Western blotting 检测细胞周期蛋白D1（cyclin D1）、p16及p19蛋白表达水平；体内用巢蛋白（Nestin）/性别决定基因高迁移率组蛋白-2（Sox2）检测两组小鼠SVZ区厚度的差异。 结果 年轻、年老小鼠的神经干细胞能表达Nestin及Sox2，符合神经干细胞的表达特性。与年轻小鼠相比，年老小鼠神经干细胞的BrdU阳性细胞比例显著下降，SA-β-Gal阳性细胞百分比显著升高，细胞活性氧水平显著升高，SVZ区厚度明显变薄，差异具有统计学意义（P<0.05）。在分子水平上表现为cyclin D1蛋白表达水平明显下降，p16、p19蛋白表达水平显著升高。 结论 年老小鼠NSCs出现增龄性变化，这些变化可能在大脑衰老中起重要作用。

关键词: 神经干细胞, 衰老, 免疫印迹法, 小鼠

Abstract:

Objective To investigate the biological changes of neural stem cells (NSCs) of mice from subventricular zone (SVZ) during aging. Methods NSCs were isolated, cultured and identified from 2 month (young) and 28 month (aged) old mice respectively, with 10 mice in each group. The proliferation ability of NSCs was determined by BrdU staining; cellular senescence was detected by senescence-associated-β-galactosidase (SA-β-gal) staining; ROS level was determined by ROS kit; expression of cyclin D1, p16, p19 protein in NSCs was tested by Western blotting. The thickness of SVZ between the two groups was detected by nestin/Sox2 staining.Results Positive expressions of neural stem cell marker nestin and Sox2 were identified in NSCs of the young and the aged mice. Aged NSCs exhibited a decreased percentage of BrdU positive cells, increased SA-β-gal activity, elevated intracellular reactive oxygen species level, decreased pool size in the SVZ zone, and there were significant differences (P<0.05). Additionally, at the molecular level, aged NSCs exhibited a decrease of cyclin D1 and an increase of p16 and p19.Conclusion Aged NSCs demonstrate aging changes which may play an important role in brain aging.

Key words: Neural stem cell, Aging, Western blotting, Mouse