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

作　　者：刘勇锋[1] 李咪咪[1] 刘芳[2] 邹奇锋 

机构地区：[1]福建医科大学附属第一医院药学部,福州350005 [2]福建医科大学附属第一医院高压氧科,福州350005 [3]莆田市第一医院药剂科,福建莆田351100

出　　处：《中国现代应用药学》2017年第2期186-190,共5页Chinese Journal of Modern Applied Pharmacy

基　　金：福建省自然科学基金资助项目(2016J01526)

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^(-1)β、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^(-1)β、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^(-1)β、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。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-1 β, 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-11）, 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-1 β, 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.

关 键 词：甲氨蝶呤 脂多糖 神经胶质细胞 pIκBα NF-ΚBP65 炎性因子 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学] R965.1[医药卫生—基础医学]