Mechanism of GSK-3 β signaling regulating CX3CL1/CX3CR1 involved in the development of migraine in rats
Objective To explore the regulatory mechanism of the interaction between trigeminal ganglion neurons and satellite glial cells(SGCs)in migraine and to identify key factors mediating this interaction.Methods The trigeminal ganglia of eight 2-to 3-day-old Wistar rats were cultured in vitro for 3 days.A migraine cell model was established using sen-sory nerve action potential(SNAP)with nitric oxide(NO)donor.The in vitro migraine simulation group was treated with SNAP at concentrations of 0.25 mmol/L,0.5 mmol/L,and 1 mmol/L for 2 hours.The inhibitor group received intervention with AR-A014418(10 µmol/L),followed by SNAP(1 mmol/L)30 min later,allowing for 2 hour intervention.The intervention-free group served as a control.The levels of calcitonin gene-related peptide(CGRP),substance P(SP),and prostaglandin E2(PGE2)in the cultured cells from each group were measured using enzyme-linked immunosorbent assay(ELISA).The positive and negative regulatory sites of glycogen synthase kinase-3β(GSK-3β)and the changes in the ex-pression of CX3CL1/CX3CR1 proteins were evaluated using immunofluorescence assay.The expression of GSK-3β and CX3CL1/CX3CR1 was semi-quantitatively analyzed using Western blotting.Results The expression levels of CGRP,PGE2,and SP in the SNAP(1 mmol/L)group were significantly higher than those in the control group,while the expres-sion levels in the inhibitor group was significantly lower than those in the SNAP(1 mmol/L)group(both P<0.05).Immu-nofluorescence assay showed that the SNAP(1 mmol/L)group exhibited decreased expression of pGSK-3β Ser9 and in-creased expression of GSK-3β Try216 and CX3CL1/CX3CR1 in trigeminal ganglion cells when compared with the control group.In contrast,the inhibitor group exhibited increased expression of pGSK-3β Ser9 and decreased expression of pGSK-3β Try216 and CX3CL1/CX3CR1 when compared with the SNAP(1 mmol/L)group.Conclusion NO enhances the ex-pression of GSK-3β in trigeminal ganglia,up-regulates CX3CL1/CX3CR1,and activates SGCs to produce inflammatory fac-tors.This signaling pathway represents a regulatory mechanism of the development and progression of migraine.
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