Clemastine promotes oligodendrocyte precursor cell differentiation and myelination after acute radiation injury

Abstract

A rat radiation injury model was constructed, in which myelin basic protein (MBP) expression was detected via western blot. Primary oligodendrocyte precursor cells (OPCs) and neuronal cells were isolated from Sprague-Dawley rats, and expression of neurofilament protein and chondroitin sulphate proteoglycan was detected via immunofluorescence to identify isolated primary cells. Cell counting kit-8 (CCK8) experiments were used to assess the effects of different concentrations of clemastine on OPC proliferation. Five groups of lentiviral shRNA vector targeting muscarinic receptor were constructed, and their transfection efficiencies were quantified using RT-qPCR and western blot. To explore the effect of clemastine on OPC differentiation and myelination, MBP and OLIG2 expression were detected via immunofluorescence and western blot. Clemastine was found to promote MBP formation in rats and inhibit OPC proliferation in a concentration-dependent manner. Compared with the control group, CHRM1, CHRM2, CHRM3, CHRM4, and CHRM5 shRNA expression significantly decreased mRNA and protein expression levels (p<0.01). Among these, CHRM2 displayed the best interference efficiency.CHRM2 interference (M2R) significantly increased MBP and OLIG2 expression in a manner that was further enhanced by clemastine treatment (p<0.05). Clemastine enhances the antimuscarinic effect, promotes the differentiation of OPCs into OLs, and plays an important role in myelination by up-regulating the expression of MBP.

Keywords

• clemastine
• cell differentiation
• myelination
• oligodendrocyte precursor cells