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| 磁刺激对小鼠原代海马神经元突触素、生长相关蛋白及脑源性神经营养因子表达的影响 |
| 马隽1,2,3,张展翅1,崔慧先1,王铭维2*,王彦永2,马芹颖2 |
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(1.河北医科大学基础医学院,石家庄 050017
2.河北省脑老化与认知神经科学实验室,石家庄 050017
3.河北体育学院,石家庄050001
*通信作者) |
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| 摘要: |
| 目的 观察磁刺激对原代海马神经元形态及突触素(SYN)、生长相关蛋白43(GAP43)、脑源性神经营养因子(BDNF)表达的影响, 探讨磁刺激对突触可塑性的影响及可能机制。方法 原代海马神经元随机分为对照组、假刺激组及40%(1 Hz,0.76 T)、60%(1 Hz,1.14 T)、80%(1 Hz,1.52 T)最大磁刺激输出强度组, 各刺激组自细胞接种后第2~6 天接受磁刺激, 连续5 d; 假刺激组置于相同磁场装置环境, 但不接受磁刺激。各组细胞于第7天相同时间取材。采用扫描电镜、细胞免疫荧光法、蛋白质印迹及RT-PCR等方法 观察神经元形态变化及SYN、GAP43与BDNF蛋白、mRNA的表达。结果 40%强度组神经元突起长度增长、胞间神经联系增多, SYN免疫反应性、BDNF免疫反应性及蛋白表达均高于对照组(P<0.05, P<0.01),GAP43免疫反应性及蛋白水平未见明显升高, SYN、GAP43与BDNF mRNA表达量高于对照组(P<0.05); 60%强度组突起长度与数量增加(P<0.01)、交织成密集网络, SYN、GAP43与BDNF免疫反应性、蛋白及mRNA表达量均高于对照组(P<0.01); 80%强度组突起数目、长度增加, 同时有细胞损伤现象, GAP43免疫反应性、BDNF免疫反应性及蛋白水平高于对照组(P<0.05,P<0.01), SYN免疫反应性及蛋白表达与对照组相比差异无统计学意义, SYN、GAP43及BDNF mRNA表达增加(P<0.05, P<0.01)。结论 磁刺激通过促进原代海马神经元BDNF的合成或分泌, 上调SYN、GAP43的表达, 影响神经元形态, 可能在调控突触可塑性、促进神经网络构建方面发挥重要作用; 不同磁刺激参数刺激效果不同。 |
| 关键词: 磁刺激 突触 神经网 原代海马神经元 |
| DOI：10.3724/SP.J.1008.2011.01096 |
| 投稿时间：2011-07-26修订日期：2011-09-16 |
| 基金项目:国家重点基础研究发展计划(“973”计划)前期研究专项(2010CB535005). |
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| Effects of magnetic stimulation on expression of SYN, GAP43, and BDNF in primary mouse hippocampal neurons |
| MA Jun1,2,3,ZHANG Zhan-chi1,CUI Hui-xian1,WANG Ming-wei2*,WANG Yan-yong2,MA Qin-ying2 |
(1. College of Basic Medical Sciences, Hebei Medical University,Shijiazhuang 050017, Hebei, China
2. Brain Aging and Cognitive Neuroscience Laboratory of Hebei Province,Shijiazhuang 050017, Hebei, China
3. Hebei Institute of Physical Education, Shijiazhuang 050001, Hebei, China
*Corresponding author.) |
| Abstract: |
| Objective To observe the effects of magnetic stimulation on the expression of synaptophysin (SYN), growth associated protein43(GAP43), brain-derived neurotrophic factor (BDNF) in primary mouse hippocampal neurons, so as to explore the possible mechanism of magnetic stimulation in mediating synaptic plasticity. Methods Primary mouse hippocampal neurons were randomly divided into control group, sham stimulation group, and magnetic stimulation groups with various output intensities: 40% (1 Hz, 0.76 T), 60% (1 Hz, 1.14 T), and 80% (1 Hz, 1.52 T) . Cells were stimulated from the 2nd d to the 6th d after plantation; cells in the sham stimulation group was placed in the same environment, but did not accept magnetic stimulation, and cells were drawn at the same time on 7th d. Scanning electron microscopy, immune cell fluorescence, Western blotting analysis and RT-PCR Methods were used to study the morphological changes of neurons and the expressions of SYN, GAP43, and BDNF at protein and mRNA levels. Results In the 40% intensity group the neurite length of neurons and neural connections between cells were both increased, and the SYN-ir, BDNF-ir COD values and protein expressions were significantly higher than those in control group(P<0.05, P<0.01), but GAP43-ir COD value and protein level were not significantly increased; SYN, GAP43, and BDNF mRNA expressions were also markedly higher than those in the control group(P<0.05). In the 60% intensity group, the length and number of neuritis (woven into dense network) were also increased significantly compared with those in the control group (P<0.01) ; COD values of immune reactivity for SYN, GAP43, and BDNF, and their expressions at protein and mRNA were all significantly higher (P<0.01). The neuritis in the 80% intensity group had increased length and number, but with cell damage; GAP43-ir and BDNF-ir COD values and protein levels were also significantly increased (P<0.05, P<0.01), but SYN-ir COD value and protein expression were not significantly different from those of the control group; and SYN, GAP43, and BDNF mRNA expressions were significantly increased compared with control group (P<0.05, P<0.01). Conclusion Magnetic stimulation may increase SYN, GAP43 expression by promoting BDNF synthesis and secretion, and thus plays a very important role in affecting neuronal morphology, regulation of synaptic plasticity, promoting neural network construction; magnetic stimulation at different parameters Results in different stimulus Results . |
| Key words: magnetic stimulation synapses nerve net primary hippocampal neurons |
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