NO对内毒素诱导的肺泡巨噬细胞核因子-κB活化和TNF-α的调节

Effect of Nitric Oxide on activation of nuclear factor-κB and production of tumor necrosis factor in lipopolysaccharide-induced pulmonary alveolar macrophages

目的 探讨一氧化氮对内毒素 (lipopolysaccharide,L PS)诱导肺泡巨噬细胞核因子 -κB(NF-κB)活化及肿瘤坏死因子α(TNF-α)基因表达的调节 .方法 用支气管肺泡灌洗法收集肺泡巨噬细胞 (pulmonary alveolar Macrophages,PAM)进行培养 ,分正常对照组 ,L PS组 ,NO+L PS组 .用凝胶电泳迁移率改变分析 (EMSA)法和 EL ISA法分别检测核提取物中 NF- κB活性和细胞培养上清中 TNF- α含量 .结果 L PS组 NF-κB活性和 TNF-α含量在刺激 2 4h后明显高于正常对照组 (P<0 .0 1) ;NO+L PS组 NF-κB活性和 TNF-α含量均显著低于 L PS组 (P<0 .0 1) .结论 L PS诱导 PAM的 NF- κB活化 ,导致 TNF- α基因表达增强 ;NO可抑制 NF-κB活化而减少 TNF-

AIM To investigate activation of nuclear factor κB (NF κB) and the expression of tumor necrosis factor α (TNF α) in lipopolysaccharide induced pulmonary alveolar Macrophages (PAM) stimulated with Nitric Oxide. METHODS PAM collected by BALT were cultured and divided into three groups: Control group, LPS stimulated group, and NO+LPS group. The NF κB activity of nuclear protein extract from the PAM and the concentration of TNF α in the supernatant were measured by electrophoretic mobility shift assay and ELISA. RESULTS The activity of NF κB and level of TNF α significantly increased 24 h after LPS stimulation; Compared with LPS stimulated group, both NF κB activity and concentration of TNF α were significantly lowered in NO+LPS group. CONCLUSION LPS might activate NF κB in the PAMs and induce the increase of transcription and expression of TNF α gene; NO could inhibit the activation of NF κB and reduce the release of TNF α.