性因子通路的影响.PDFVIP

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  1a 1a* 1b 2 刘勇锋 ，李咪咪 ，刘芳 ，邹奇锋 (1.福建医科大学附属第一医院，a.药学部，b.高压氧科，福州 350005 ；2.莆田市第一医 院药剂科，福建 莆田 351100) 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(P0.05 或P0.01) 。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(P0.05) 。 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。 关键词：甲氨蝶呤；脂多糖；神经胶质细胞；pIκBα；NF-κBp65 ；炎性因子 中图分类号：R322.81 ；R965.1 文献标志码：A 文章编号：1007-7693(2017)02-0186-05 DOI: 10.13748/ki.issn1007-7693.2017.02.008 Effect of Methotrexate on the Pathway of LPS-induced pIκBα-NF-κBp65-Inflammatory Cytokine in the Rat Spinal Cord Glial Cells 1a 1a* 1b 2 LIU Yongfeng , LI Mimi , LIU Fang , ZOU Qifeng (1.The First Affiliated Hospital of Fujian Medical University, a. The Department of Pharmacy, b.The Department of Hyperbaric Oxygen, Fuzhou 350005, China; 2.The Department of Pharmacy, The First Hospital of Putian City, Putian 351100, China) 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(includ