远志皂苷元促进人神经干细胞体外增生和分化的研究

doi:10.3969/j.issn.1006-7795.2013.04.016

首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (4): 559-565.doi: 10.3969/j.issn.1006-7795.2013.04.016

远志皂苷元促进人神经干细胞体外增生和分化的研究

师昉^1,2, 梁志刚¹, 郭紫瑄¹, 汪璇¹, 王晓民¹

1. 首都医科大学神经生物学系, 教育部神经变性病重点实验室, 北京 100069;
2. 北京市残疾人辅助器具资源中心, 北京 100028

收稿日期:2013-02-16 出版日期:2013-08-21 发布日期:2013-07-20
通讯作者: 王晓民 E-mail:xmwang@ccmu.edu.cn
基金资助:
国家重点基础研究发展计划项目(973项目)(2011CB504100);国家自然科学基金重点项目(81030062);北京市自然科学基金(5102010)。

Senegenin promotes in vitro proliferation and differentiation of human neural progenitor cells

SHI Fang^1,2, LIANG Zhigang¹, GUO Zixuan¹, WANG Xuan¹, WANG Xiaomin¹

1. Department of Neurobiology, Capital Medical University, Key Laboratory for Neurodegenerative Disorders, Ministry of Education, Beijing 100069, China;
2. Resource Center of Beijing Assistant Technology, Beijing 100028, China

Received:2013-02-16 Online:2013-08-21 Published:2013-07-20
Supported by:
This study was supported by National Basic Research Program of China(973 Project)(2011CB504100), National Natural Science Foundation of China(81030062), Natural Science Foundation of Beijing(5102010).

摘要/Abstract

摘要：

目的研究远志皂苷元对体外培养的人神经干细胞增生和分化的影响及其可能的机制。方法对体外培养的人神经干细胞系进行细胞鉴定;在培养液中分别加入50 μmol/L和5 μmol/L的远志皂苷元,同时设置空白对照组,用CCK8试剂在酶标仪上进行细胞吸光度测试用以检测细胞活力;利用实时荧光定量RT-PCR方法分别检测50 μmol/L和5 μmol/L的远志皂苷元对神经干细胞负性分化调节基因Hes1和正性分化调节基因Mash1的mRNA表达影响;免疫荧光染色标记法标记分化后的神经元细胞和星形胶质细胞的比例。结果与空白对照组比较,50 μmol/L远志皂苷元可显著性增加神经干细胞数目(P<0.05),上调Hes1基因表达(P<0.05);5 μmol/L远志皂苷元也可增加神经干细胞数目,同时上调了Hes1和Mash1基因表达(P<0.05);5 μmol/L远志皂苷元可以促进神经干细胞分化,提高神经元分化的比例(P<0.05)。结论远志皂苷元可以促进体外培养的神经干细胞增生和分化,这种促增生和分化的作用可能是通过上调Hes1和Mash1基因表达实现的。

关键词: 远志皂苷元, 神经干细胞, 增生, 分化

Abstract:

Objective To study the effects of senegenin on the proliferation and differentiation of human neural stem cells in vitro and its possible mechanism.Methods Human neural stem cell line was cultured and identified in vitro. They were incubated with 50 μmol/L or 5 μmol/L senegenin respectively, and the group added with the vehicle was set as the control. Then the cell absorbance was measured by CCK8 assay on a microplate reader to test the cell viability. The effects of 50 μmol/L or 5 μmol/L senegenin on the mRNA expression of negative differentiation regulating gene Hes1 and positive differentiation regulating gene Mash1 were examined by real-time fluorescence quantitative RT-PCR. The differentiation ratio of neurons and astrocytes from the neural stem cells was assessed through immune-fluorescence staining process.Results Senegenin at 50 μmol/L significantly increased the cell number compared with the control group(P<0.05), and upregulated the Hes1 mRNA expression(P<0.05). Senegenin at 5 μmol/L also promoted the cell proliferation, and at the same time it increased the mRNA expressions of both Hes1 and Mash1 apparently(P<0.05). Moreover, 5 μmol/L senegenin obviously promoted the neuronal differentiation from the neural stem cells. Conclusion Senegenin can promote the proliferation and differentiation of human neural stem cells in vitro, and this effect may be induced by the up-regulation of Hes1 and Mash1 gene expression.

Key words: senegenin, human neural stem cells, proliferation, differentiation

中图分类号:

R338

师昉, 梁志刚, 郭紫瑄, 汪璇, 王晓民. 远志皂苷元促进人神经干细胞体外增生和分化的研究[J]. 首都医科大学学报, 2013, 34(4): 559-565.

SHI Fang, LIANG Zhigang, GUO Zixuan, WANG Xuan, WANG Xiaomin. Senegenin promotes in vitro proliferation and differentiation of human neural progenitor cells[J]. Journal of Capital Medical University, 2013, 34(4): 559-565.

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远志皂苷元促进人神经干细胞体外增生和分化的研究

Senegenin promotes in vitro proliferation and differentiation of human neural progenitor cells

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