| 刘军昌,薛姗姗,周翠红,王化宁,何 宏.调节TREK1对小鼠海马神经干细胞增殖和BDNF表达的影响[J].,2019,19(7):1227-1232 |
| 调节TREK1对小鼠海马神经干细胞增殖和BDNF表达的影响 |
| Regulation of TREK1 on the Proliferation and BDNF Expression in the Hippocampal Neural Stem Cells of Mice |
| 投稿时间：2018-09-23 修订日期：2018-10-18 |
| DOI：10.13241/j.cnki.pmb.2019.07.006 |
| 中文关键词: 神经干细胞 增殖 TREK-1 BDNF |
| 英文关键词: Neural stem cells Proliferation TREK-1 BDNF |
| 基金项目:国家自然科学基金项目（81571309）；陕西省重点研发计划项目（2017ZDXM-SF-047） |
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| 中文摘要: |
| 摘要 目的：观察调节TWIK相关的K+通道1（TREK1）对小鼠海马神经干细胞增殖和脑源性神经营养因子（brain derived neu- rotrophic factor，BDNF）表达的影响。方法：从孕10.5 d C57bl/6J小鼠胚胎海马中分离出神经干细胞并培养，待细胞达到70% ~ 80%融合后，对细胞进行慢病毒干预，细胞分为sham组，Ctrl组（转染对照病毒），Plenti-TREK-1组（转染携带TREK1高表达载体病毒）和sh-TREK-1组（转染携带TREK1-shRNA病毒）。采用CCK-8法检测病毒干预后3天和7天各组神经干细胞的细胞活力，采用qRT-PCR检测病毒干预后7天各组神经干细胞TREK-1基因表达情况，并通过Elisa检测各组神经干细胞上清液BDNF表达水平。结果：与sham组相比较，（1）Plenti-TREK-1组TREK1基因表达上调，神经干细胞的神经球体积减小，细胞活力降低，细胞增殖减少，细胞上清BDNF水平下降；（2）sh-TREK-1组TREK1基因表达下调，神经干细胞的神经球体积增加，细胞活力增高，细胞增殖增加，细胞上清BDNF水平上调；（3）上述各项指标Ctrl组与sham组之间无统计学差异。结论：神经干细胞的增殖与TREK1通道密切相关，上调TREK-1可以抑制神经干细胞增殖及其BDNF的表达水平；下调TREK-1则可以促进神经干细胞增殖并上调其BDNF的表达水平。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of TREK-1 over-express or knock-down on the proliferation and brain derived neurotrophic factor (BDNF) expression in neural stem cells from the mice hippocampus. Methods: Neural stem cells were isolated from the embryonic hippocampus of 10.5 D C57bl/6J mice and cultured. After the cells reached 70% ~ 80% fusion, the cells were subjected to lentiviral intervention and divided into sham group, Ctrl group, Plenti-TREK-1 group and sh-TREK-1 group. The cell viability in each group was detected by CCK-8 method at 3 and 7 days after virus intervention. The expression of TREK-1 gene and BDNF in the super- natant was detected by QRT-PCR or Elisa at 7 days after virus intervention respectively. Results: (1) Compared with sham group, the mRNA level of TREK1 was up-regulated and the volume of neurospheres, cell activity and cell proliferation as well as the level of BDNF in the supernatant were reduced in Plenti-TREK-1 group; (2) the mRNA level of TREK1 was down-regulated and the volume of neuro- spheres, cell activity and cell proliferation as well as the level of BDNF in the supernatant were increased in sh-TREK-1 group when compared with sham group; (3) There were no significant differences between sham and Ctrl group in the above parameters. Conclusion: The proliferation of neural stem cells is mediated by the expression of TREK1 channel. Up-regulation of TREK-1 inhibits the prolifera- tion of neural stem cells and the expression of BDNF, and down-regulation of TREK-1 promotes the proliferation of neural stem cells and up-regulate the expression of BDNF. |
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