解剖学报

解剖学报 ›› 2018, Vol. 49 ›› Issue (4): 424-430.doi: 10.16098/j.issn.0529-1356.2018.04.002

• 神经生物学 • 上一篇    下一篇

干预脑脂质结合蛋白表达对神经干细胞分化作用的影响

于婷婷 李浩明 金国华* 韩笑 秦建兵   

  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:2017-04-28 修回日期:2017-06-11 出版日期:2018-08-06 发布日期:2018-08-06
  • 通讯作者: 金国华 E-mail:jguohua@ntu.edu.cn
  • 基金资助:
    江苏省自然科学基金青年项目;南通大学研究生创新计划项目

Effects of intervention of brain lipid binding protein expression on differentiation of neural stem cells

YU Ting-ting LI Hao-ming JIN Guo-hua* HAN Xiao QIN Jian-bing   

  1. Department of Anatomy，Jiangsu Key Laboratory of Neuroregeneration，School of Medicine，Nantong University，Jiangsu Nantong226001，China
  • Received:2017-04-28 Revised:2017-06-11 Online:2018-08-06 Published:2018-08-06
  • Contact: JIN Guo-hua E-mail:jguohua@ntu.edu.cn

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摘要：

目的 探讨脑脂结合蛋白（BLBP）在大鼠海马神经干细胞（NSCs）向神经元分化中的作用。方法 构建BLBP过表达腺病毒和干扰慢病毒载体，并从大鼠胚胎海马组织中分离培养神经干细胞，采用Nestin免疫荧光鉴定NSCs;将体外培养的NSCs分为4组：腺病毒阴性对照组（Ad-NC组），BLBP过表达腺病毒组（Ad-BLBP组），慢病毒阴性对照组（LV-NC-RNAi组）和BLBP干扰慢病毒感染组(LV-BLBP-RNAi组)。Real-time PCR和Western blotting检测各组细胞中BLBP表达；病毒感染4 d后免疫荧光检测分化细胞中βⅢ微管蛋白（Tuj1）阳性神经元数量。结果 Ad-BLBP组较Ad-NC组NSCs分化为神经元的数量少，突起短而少；LV-BLBP-RNAi组较LV-NC-RNAi组NSCs分化为神经元的数量明显增多，突起多而长。结论  下调BLBP的表达能够促进体外培养的NSCs向神经元分化。

关键词: 脑脂质结合蛋白, 神经干细胞, 分化, 神经元, 免疫印迹法, 大鼠

Abstract:

Objective  To explore the effect of BLBP on differentiation of hippocampal neural stem cells (NSCs) into neurons in vitro. Methods  BLBP overexpression adenovirus and BLBP RNA interference lentiviral vector were built. NSCs were obtained from rat hippocampus by serum-free culturing and immunofluorescence was used to identify NSCs. The NSCs were assigned to Ad-NC group, Ad-BLBP group, LV-NC-RNAi group and LV-BLBP-RNAi group. On the 4th day after infection, BLBP mRNA and protein expression in the NSCs were detected by Real-time PCR and Western blotting, and the neuronal class Ⅲ β-tubulin (Tuj1) positive cells were detected by immunofluorescence. Results  NSCs in Ad-BLBP group expressed a high level of BLBP and there were only fewer Tuj1 positive cells than the Ad-NC group; NSCs in LV-BLBP-RNAi group expressed a lower level of BLBP and there were more Tuj1 positive cells than the LV-NC-RNAi group. Conclusion  Down regulation of BLBP expression can promote the differentiation of NSCs into neurons in vitro.

Key words: Brain lipid binding protein, Neural stem cells, Differentiation, Neurons, Western blotting, Rat