电刺激对背根神经节和雪旺细胞髓鞘化的影响

Effects of electrical stimulation on the myelination of dorsal root ganglia and Schwann cells

目的:建立体外的背根神经节(DRG)—雪旺细胞(SCs)—电刺激模型,为研究电刺激促周围神经成髓鞘机制提供基础。方法:以函数发生器和电刺激小室构建电刺激细胞培养系统,DRG/SCs共培养24 h后分为对照(Ctrl)组和电刺激(ES)组。ES组给予矩形波电刺激,6 Vp-p,10 Hz,1 h/d,7 d。Ctrl组无电刺激。利用CCK8观测电刺激对细胞毒性的影响,利用髓磷脂碱性蛋白(MBP)免疫荧光检测电刺激对DRG/SCs髓鞘化的影响。结果:CCK8细胞增殖毒性实验显示,ES组光密度值略高于Ctrl组,但两组之间无统计学意义。免疫荧光检测结果显示ES组MBP的荧光强度明显高于Ctrl组(P<0.01)。结论:本文设计的电刺激细胞培养系统对神经细胞安全无毒,连续7 d的电刺激可提高神经髓鞘化率。

Objective To establish an in vitro dorsal root ganglion(DRG)-Schwann cells(SCs)-electrical stimulation(ES)model for providing a basis for the investigation on the mechanism of ES in promoting peripheral nerve myelination.Methods A function generator and an ES chamber were used to construct an ES cell culture system.After being co-cultured for 24 h,DRG/SCs were divided into control group and ES group.The ES group was given rectangular wave ES,6 Vp-p,10 Hz,1 h/d,for 7 days;and no ES was applied in control group.The effect of ES on cytotoxicity was observed using CCK8,and the effect of ES on the myelination of DRG/SCs was detected by immunofluorescence using myelin basic protein.Results The CCK8 cell proliferation and toxicity assay showed that the optical density of ES group was slightly higher than that of control group,but there was no statistical significance between two groups.The immunofluorescence assay revealed that the fluorescence intensity of myelin basic protein in ES group was significantly higher than that in control group(P<0.01).Conclusion The ES cell culture system designed in the study is safe and non-toxic to neuronal cells,and ES for 7 days in succession can increase the rate of neural myelination.