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    甲氨蝶呤对脂多糖诱导的大鼠脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路的影响

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

      摘要
    • 摘要: 目的 观察甲氨蝶呤对脂多糖（lipopolysaccharide，LPS）诱导的大鼠脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路的影响。方法 脊髓组织块法培养神经胶质细胞。将分离的神经胶质细胞接种于多孔板培养48 h后，分为空白对照组、LPS组、LPS+pIκBα抑制剂组、LPS+甲氨喋呤组。随后应用免疫印迹法测定各组分的pIκBα与胞核及胞浆NF-κBp65水平变化，酶免疫法（ELISA）测定炎性因子TNF-α、IL-1β、IL-6含量。结果 神经胶质细胞经LPS诱导后，pIκBα、胞核NF-κBp65和细胞上清液炎性因子TNF-α、IL-1β、IL-6水平均显著增加（P<0.05或P<0.01）。甲氨喋呤可明显抑制经LPS诱导的神经胶质细胞pIκBα水平，显著降低胞核NF-κBp65水平和细胞上清液炎性因子TNF-α、IL-1β、IL-6的含量（P<0.05）。结论 甲氨喋呤对LPS诱导的脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路有显著的抑制作用。

       

      Abstract: 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.

       

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