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山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (4): 49-54.doi: 10.6040/j.issn.1671-7554.0.2014.818

• 基础医学 • 上一篇    下一篇

茶氨酸对神经干细胞增殖和分化的影响

梁宇1, 辛涛1, 刘倩2, 郑向荣1, 杨依航1, 庞琦1   

  1. 1. 山东大学附属省立医院神经外科, 山东 济南 250021;
    2. 山东大学医学院组织胚胎学教研室, 山东 济南 250012
  • 收稿日期:2014-11-12 修回日期:2015-03-09 出版日期:2015-04-10 发布日期:2015-04-10
  • 通讯作者: 庞琦.E-mail:pangqi@sdu.edu.cn E-mail:pangqi@sdu.edu.cn
  • 基金资助:
    国家自然科学基金(81171062, 81100836);山东省科技发展计划(2014GSF118025)

Effects of theanine on the proliferation and differentiation of neural stem cells

LIANG Yu1, XIN Tao1, LIU Qian2, ZHENG Xiangrong1, YANG Yihang1, PANG Qi1   

  1. 1. Department of Neurosurgery, Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong, China;
    2. Department of Histology and Embryology, School of Medicine, Shandong University, Jinan 250012, Shandong, China
  • Received:2014-11-12 Revised:2015-03-09 Online:2015-04-10 Published:2015-04-10

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摘要： 目的 探讨茶氨酸对神经干细胞增殖和分化的影响.方法 采用机械吹打结合200目钢网滤过的方法,从12.5 d的昆明胎鼠大脑中分离NSCs并培养;用巢蛋白(nestin)免疫荧光染色鉴定NSC;加入不同浓度茶氨酸(theanine),MTS法检测不同时间点活性,选定茶氨酸最终加药浓度(100 μmol/L);将NSCs分为两组:对照组、加药组(茶氨酸浓度为100 μmol/L);培养后进行BrdU染色和微管相关蛋白2(MAP2)、胶质纤维酸性蛋白(GFAP)免疫荧光染色;利用Q-PCR以及Westen blotting检测MAP2和GFAP表达.结果 100 μmol/L茶氨酸处理组MTS检测的吸光度值显著增高(P <0.05);加药组BrdU阳性细胞比率、MAP2阳性细胞比率明显高于对照组(P <0.05),两组GFAP阳性细胞比率无统计学差异;加药组MAP2表达明显高于对照组(P <0.05),两组GFAP表达无统计学差异.结论 茶氨酸可促进NSCs增殖及其向神经元方向分化.

关键词: 茶氨酸, 健康教育, 依从性, 功能性消化不良, 分化, 小鼠, 增殖, 神经干细胞, 焦虑

Abstract: Objective To explore the effects of theanine on the proliferation and differentiation of neural stem cells (NSCs). Methods NSCs were isolated from Kunming mouse embryonic at E12.5 by mechanical blowing combined with stencils of 200 mesh. Isolated NSCs were then identified with nestin immunofluorescent staining technique. Different concentrations of theanine were added into nutrient fluid, cell activity at different time was detected with MTS, and the final concentration of theanine (100 μmol/L) was determined. Then NSCs were divided into two groups: control group, theanine (100 μmol/L) treated group. The proliferation and differentiation of NSCs were determined with BrdU labeling and immunofluorescent staining of microtubule associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP). The Q-PCR and Western blotting were used to detect the expression of MAP2 and GFAP. Results The optical density of theanine (100 μmol/L) treated group was significantly increased in MTS (P <0.05). The ratio of BrdU-positive cells and MAP2-positive cells against total NSCs treated with theanine was significantly increased in comparison with that in control group (P <0.05), but there was no noticeable difference in GFAP immunofluorescence between the two groups. The expression of MAP2 treated with theanine was significantly increased compared with that in the control group, there was no noticeable difference in GFAP expression between the two groups. Conclusion Theanine (100 μmol/L) can promote the proliferation of NSCs and its neuronal differentiation.

Key words: Proliferation, Differentiation, Mice, Theanine, Neural stem cells

中图分类号: 

  • R34
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