In Vitro Study On Promotion Of Oligodendrocyte Progenitor Transformation And Re-myelination By Neuregulin-1/ErbB4 Mediated Electrical Stimulation

Objective(s):To explore the changes of electrical stimulation parameters of re-myelination and the expression of basic myelin protein(MBP)in a co-culture model of neuron/oligodendrocyte re-myelination promoted by electrostimulation(ES)in vitro;To explore the molecular mechanism of electrical stimulation promoting re-myelination of in vitro co-culture system,so as to provide more theoretical basis for the clinical treatment of electrical stimulation.Methods:(1)Primary neurons and oligodendrocyte progenitors were extracted from SD rats within 24 hours of birth,and Neurobasal/B27/N2 medium was used for co-culture of primary neurons/oligodendrocyte progenitors.(2)Electrical stimulation intervention was performed on the co-culture model from three aspects of action events,stimulus intensity and stimulus frequency.After cell collection,the expression level of MBP was analyzed by Western Bloting,and effective parameters were screened for further experiments.(3)P0 was used to label SCs-like cells and PLP was used to label mature OLs cells to observe whether electrical stimulation promoted proliferation and transformation of SCs-like cells and OLs cells.(4)Exogenous NRG-1 was added into the medium to observe the changes in the expressions of MBP,P0 and PLP in the neuron-oligodendrocyte myelination co-culture model,and the changes in the number of SCs-like cells and OLs were recorded.(5)After adding Erb B4 receptor antagonist PD158780 into the medium,electrical stimulation was performed to observe the changes in the expressions of MBP,P0 and PLP in the neuron-oligodendrocyte myelination co-culture model,and the changes in the number of SCs-like cells and OLs were recorded.Results:(1)After 2W of electrical stimulation,the combination of electrical stimulation parameters of 10min/d,intensity of 10,frequency of 10 Hz could effectively promote the expression of MBP;(2)Electrical stimulation can increase the number of SCs-like and the expression of P0 in neuron-oligodendrocyte myelination coculture model;(3)Exogenous NRG-1 can promote the expression of MPB and P0 and increase the number of SCs-like cells in neuron-oligodendrocyte myelination co-culture model.(4)Erb B4 receptor antagonist PD158780 can reduce the expressions of MBP and P0 and reduce the number of Scs-like in the co-culture system after electrical stimulation.Conclusion(s):(1)ES can promote the expression of MBP in co-culture models,possibly mediated by SCs-like cells.(2)ES can activate the expression of MBP in NRG-1/Erb B4 signaling pathway and increase the number of SCs-like cells.