Effect of Methotrexate on the Pathway of LPS-induced pIκBα-NF-κBp65-Inflammatory Cytokine in the Rat Spinal Cord Glial Cells

OBJECTIVE To study the effect of methotrexate on the pathway of lipolysaccharide(LPS)-induced pIκBα-NF-κBp65-inflammatory cytokine in glial cells of the rat spinal cord.METHODS The glial cells were obtained from the spinal cord tissue nubbles, then were inoculated into multi-well plate and cultured for 48 h. Cells were divided into 4 groups:control group, LPS group, LPS+pIκBα inhibitor group and LPS+methotrexate group. The expression of pIκBα, NF-κBp65 in each groups were detected by Western blot, and the concentrations of inflammatory factors(including TNF-α, IL^-1β, IL-6) were measured by ELISA.RESULTS The LPS group showed significantly higher expressions of pIκBα, NF-κBp65 protein in the nucleus and the concentrations of inflammatory factors(including TNF-α, IL^-1β, IL-6) compared to the normal control group (P<0.05 or P<0.01). Compared with LPS group, the expressions of pIκBα, NF-κBp65 protein and concentration of TNF-α, IL^-1β, IL-6 were significantly decreased in LPS plus methotrexate group and pIκBα inhibitor group(P<0.05).CONCLUSION Methotrexate can markedly inhibit the pathway of LPS-induced pIκBα-NF-κBp65-inflammatory cytokines in glial cells.