解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (5): 627-632.doi: 10.3969/j.issn.0529-1356.2014.05.008

• 神经生物学 • 上一篇    下一篇

雌二醇对大鼠海马神经干细胞增殖的影响

刘晓东 张夏南 郝宁 鞠前前 秦建兵 田美玲 金国华 吕广明*   

  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:2014-02-19 修回日期:2014-05-23 出版日期:2014-10-06 发布日期:2013-10-06
  • 通讯作者: 吕广明 E-mail:gmlu@ntu.edu.cn
  • 基金资助:

    江苏高校优势学科建设项目

Effect of estradiol on proliferation of rat hippocampal neural stem cells

LIU Xiao-dong ZHANG Xia-nan HAO Ning JU Qian-qian QIN Jian-bing TIAN Mei-ling JIN Guo-hua LÜ Guang-ming*   

  1. Department of Human Anatomy, Medical School of Nantong University, Jiangsu Key Laboratory ofNeuroregeneration, Jiangsu Nantong 226001, China
  • Received:2014-02-19 Revised:2014-05-23 Online:2014-10-06 Published:2013-10-06
  • Contact: Lü Guang-ming E-mail:gmlu@ntu.edu.cn

摘要：

目的 观察雌二醇对大鼠海马神经干细胞增殖的影响及其作用机制。方法 取20只孕17d的SD大鼠海马组织，进行神经干细胞的培养和增殖，添加不同浓度的雌二醇（E2）。通过5-溴脱氧尿嘧啶核苷(BrdU)免疫荧光、MTT方法检测神经干细胞的增殖；通过免疫荧光技术和巢蛋白(Nestin)双标观察神经干细胞球中雌激素受体ERα和ERβ的表达情况。结果 BrdU与MTT检测结果显示，随着雌二醇浓度从10-10mol/L增加至10-8mol/L，神经干细胞数量逐渐增加。雌二醇在10-8 mol/L时，细胞增殖数目最多而且细胞活力最好。随着雌二醇浓度进一步增加至10-6mol/L，神经干细胞增殖能力逐渐下降。免疫荧光技术检测显示神经干细胞均表达ERα和ERβ两种受体。 结论 在一定浓度范围内，雌二醇能促进海马神经干细胞的增殖，并可能是通过ERα和ERβ介导促进神经干细胞增殖。

关键词: 海马, 神经干细胞, 雌二醇, 增殖, 雌激素受体, 免疫荧光, 大鼠

Abstract:

Objective The aim is to observe the role and mechanism of estradiol (E2) on the proliferation of rat hippocampal neural stem cells (NSCs). Methods Twenty hippocampi from embryonic 17-day (E17) SD rats were dissociated and plated into culture flasks with NSCs specific medium containing different concentrations of estradiol. The proliferation and the vitality of NSCs were detected by immunofluorescence against BrdU and MTT assay. The expression of estrogen receptors (ERα and ERβ) was measured by immunofluorescence staining combined with Nestin double labeling. Results BrdU and MTT assay results showed that the cell number increased when the concentration of estradiol increased from 10-10 to 10-8 mol/L. The number of cell proliferation and the viability of cells were best at the concentration of 10-8 mol/L compared to the other groups. However, when the estradiol concentration was increased from 10-8 to 10-6 mol/L, the cell proliferative capacity declined gradually. Double immunofluorescence labeling showed that the two types of estrogen receptors (ERα and ERβ) were expressed in the cultured hippocampal NSCs. Conclusion Estradiol promotes the proliferation of hippocampal NSCs in a certain concentration range, and ERα and ERβ may be involved in the estradiol-induced proliferation.

Key words: Hippocampus, Neural stem cells；Estradiol, Proliferation；Estradiol receptor, Immunofluorescence, Rat