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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%(1Hz,0.76T)、60%(1Hz,1.14T)、80%(1Hz,1.52T)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5d;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。
Objective To observe the effects of magnetic stimulation on the morphous of primary hippocampal neurons and the expression of synapse (SYN), growth associated protein 43 (GAP43) and brain derived neurotrophic factor (BDNF) and to explore the effect of magnetic stimulation on synaptic plasticity Possible mechanism. Methods Primary cultured hippocampal neurons were randomly divided into control group, sham stimulation group and 40% (1Hz, 0.76T), 60% (1Hz, 1.14T), 80% (1Hz, 1.52T) The stimulation group received magnetic stimulation from day 2 to day 6 after inoculation for 5 days continuously. The sham stimulation group was placed in the same magnetic field device environment but did not receive magnetic stimulation. Each group of cells in the same time on the seventh day drawing. The morphological changes of neurons and the expression of SYN, GAP43 and BDNF protein and mRNA were observed by scanning electron microscopy, immunofluorescence, Western blot and RT-PCR. Results Compared with the control group, the immunoreactivity and protein expression of GAP43 were significantly increased (P <0.05, P <0.01), the neurite outgrowth in the 40% The levels of SYN, GAP43 and BDNF mRNA were higher than those of the control group (P <0.05). The length and number of the protrusions in the 60% strength group were increased (P <0.01) (P <0.01). The number and length of neurites in 80% intensity group increased with cell injury, GAP43 immunoreactivity, BDNF immunoreactivity and protein level were higher than those in control group Compared with the control group, the expression of SYN, GAP43 and BDNF mRNA increased (P <0.05, P <0.01) in the control group (P <0.05, P <0.01). There was no significant difference in the SYN immunoreactivity and protein expression between the two groups. Conclusion Magnetic stimulation plays an important role in the regulation of synaptic plasticity and neural network construction by promoting the synthesis or secretion of BDNF in primary hippocampal neurons, up-regulating the expression of SYN and GAP43, and affecting neuronal morphology. The effect of different stimulation parameters different.