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目的:探讨体外不同条件的磁刺激对神经干细胞细胞增殖、细胞周期、细胞凋亡和细胞分化等方面的影响。方法:对离体人胚神经干细胞进行磁刺激处理,采用频率0.5 Hz,脉冲刺激波宽为72μs,刺激强度为阈上刺激1.44 T,每天1次,连续刺激3 d。按刺激次数分为A组(30个脉冲刺激/次)、B组(60个脉冲刺激/次)、C组(90个脉冲刺激/次)、D组(对照组,不进行磁刺激处理);用四甲基偶氮唑盐(MTT)法检测磁刺激对神经干细胞增殖的作用;用流式细胞术检测磁刺激对神经干细胞的细胞周期、细胞凋亡和细胞分化的作用。结果:A、B、C组神经干细胞在接受磁刺激后24～48 h的D值较D组明显增高(P<0.05),提示磁刺激对神经干细胞有轻度促增殖作用;A、B、C组G0/G1期细胞比例稍低于D组,而相应的G2/M比例较高,但无统计学差异;A、B、C组β-tuberlin阳性细胞比例较D组高,神经元比例由21.70%上升至34.17%(P<0.05)。结论:磁刺激在轻度促进细胞增殖的条件下,可诱导神经干细胞向神经元方向分化,有利于神经功能重建。
Objective: To investigate the effects of magnetic stimulation in vitro on the proliferation, cell cycle, apoptosis and cell differentiation of neural stem cells. Methods: Human embryonic neural stem cells (NSCs) were stimulated by magnetic stimulation at a frequency of 0.5 Hz with a pulse width of 72 μs and a stimulus intensity of 1.44 T at the top of the threshold, once a day for 3 days. The rats were divided into three groups according to the number of stimulation: group A (30 pulses), group B (60 pulses), group C (90 pulses), group D (control group without magnetic stimulation) The effect of magnetic stimulation on the proliferation of neural stem cells was detected by MTT assay. The effect of magnetic stimulation on the cell cycle, apoptosis and cell differentiation of neural stem cells was detected by flow cytometry. Results: The D values of neural stem cells in group A, B and C increased significantly from 24 to 48 h after magnetic stimulation (P <0.05), suggesting that magnetic stimulation had a slight effect on proliferation of neural stem cells; A, B, The proportion of cells in G0 / G1 phase in group C was slightly lower than that in group D, while the corresponding proportion of G2 / M was higher but there was no statistical difference. The proportion of β-tuberlin positive cells in group A, B and C was higher than that in group D, From 21.70% to 34.17% (P <0.05). CONCLUSION: Magnetic stimulation can induce neural stem cells to differentiate into neurons under mild stimulation of cell proliferation, which is good for neural function reconstruction.