| 李伟坤,白文芳,许伟成,白利明,李新平,朱洪翔,张 冲,李媚慧,刘晓涵,张鸣生.50Hz电磁场促进骨髓源神经祖细胞向神经元分化作用的研究[J].中国康复医学杂志,2016,(7):723~728 |
| 50Hz电磁场促进骨髓源神经祖细胞向神经元分化作用的研究 点此下载全文 |
| 李伟坤 白文芳 许伟成 白利明 李新平 朱洪翔 张 冲 李媚慧 刘晓涵 张鸣生 |
| 暨南大学第一临床医学院，广州，510630 |
| 基金项目：国家青年自然科学基金项目（81301680）；广东省科技计划项目（2012B061700059）；广东省科技厅科技发展专项资金项目（2016A020214014） |
| DOI： |
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| 摘要： |
| 摘要
目的：通过分析50Hz电磁场干预骨髓源神经祖细胞向神经细胞分化前后相关神经细胞标志物mRNA表达水平，探讨低频电磁场对骨髓源神经祖细胞诱导分化的作用。
方法：利用全骨髓培养法获取骨髓间充质干细胞，将第2代骨髓间充质干细胞在无血清神经干细胞培养环境中悬浮诱导，获取骨髓源神经祖细胞。将第3代骨髓源神经祖细胞分为2组贴壁培养：电磁场组与对照组。电磁场组干预方法为正弦波磁场、频率50Hz、强度5mT，60min/d，共15天，对照组置于无磁场干预的同等环境中。骨髓源神经祖细胞诱导后，采用免疫细胞荧光检测巢蛋白（Nestin）与微管蛋白抗体（Tuj-1）的表达变化；采用实时定量基因扩增荧光检测系统（Q-PCR）检测Nestin，唾液酸-神经细胞粘附分子（PSA-NCAM）和β-微管蛋白-Ⅲ（β-Ⅲ tubulin），乙酰胆碱酯酶（ACHE），5-羟色胺（5-HT），γ-氨基丁酸（GABA）mRNA表达水平变化。
结果：骨髓间充质干细胞在无血清神经干细胞培养环境中可形成骨髓源神经祖细胞，表达Nestin阳性产物；两组诱导后的神经元样细胞免疫细胞荧光检测Tuj-1均呈阳性表达，Q-PCR结果示Nestin，PSA-NCAM，β-Ⅲ tubulin，ACHE，5-HT，GABA mRNA表达水平与诱导前比较显著下降（P＜0.01），电磁场组β-Ⅲ tubulin mRNA表达水平显著高于对照组（P＜0.05）。
结论：50Hz电磁场可以促进骨髓源神经祖细胞向神经元分化。 |
| 关键词：骨髓间充质干细胞 骨髓源神经祖细胞 神经细胞 低频电磁场 分化 |
| 50Hz electromagnetic fields facilitate bone marrow stroma cells-derived neural progenitor cells to differentiate into neurons Download Fulltext |
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| The First School of Clinical Medicine, Jinan University, Guangzhou, 510632 |
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| Abstract: |
| Abstract
Objective: By analyzing the mRNA expression level of nerve cell markers and neurotransmitters before and after the intervention of 50Hz electromagnetic fields, we investigated the effects of low frequency electromagnetic fields on the differentiation ability of bone marrow-derived neural progenitor cells (BM-NPCs)differentiation ability.
Method: Firstly, bone mesenchymal stem cells (BMSCs) were gained by the whole bone marrow culture method, and the second generation BMSCs were cultured and induced under neural stem cells (NSCs) culture conditions to obtain the BM-NPCs. Secondly, the third generation BM-NPCs were randomly divided into two groups: an electromagnetic field group and a control group. The electromagnetic field group was exposured to a sine wave electromagnetic fields with the frequency of 50Hz and magnetic induction of 5mT, and 60min per day for fifteen days. The control group was treated under the same condition without exposure. After BM-NPCs induced differentiation, the cell marker of Nestin and Tuj-1 were analyzed by immunofluorescence and the mRNA expression of Nestin, PSA-NCAM, β-III tubulin, ACHE, GABA and 5-HT were tested by real-time quantitative reverse transcriptase polymerase chain reaction method.
Result:Under serum-free neural stem cells suspension culture conditions, the BMSCs could be aggregated into BM-NPCs with positive Nestin. After the induction of BM-NPCs, the results of immunofluorescence showed the positive expression of Tuj-1 for neuron-like cells in two groups. After induction, the mRNA expression of Nestin, PSA-NCAM, β-Ⅲ tubulin, ACHE, GABA and 5-HT was significantly decreased (P＜0.01). Further more, the β-Ⅲ tubulin mRNA level in the electromagnetic field group was higher than in the control group (P＜0.05).
Conclusion: 50Hz electromagnetic fields could induce neural progenitor cells differentiation into neurons. |
| Keywords:bone mesenchymal stem cells bone marrow-derived neural progenitor cells neurons low-frequency electromagnetic fields differentiation |
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