白介素-10对内毒素诱导肺泡巨噬细胞核因子-κB活化及肿瘤坏死因子释放的调节

摘要

Objective 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 Interleukin-10.Methods PAM collected by BALT were cultured and divided into three groups:control group;LPS-stimulated group;IL-10＋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 (EMSA) and ELISA. Results The activity of NF-κB and the level of TNF-α significantly increased from 0.5 to 4 hour after LPS stimulation;Compared with LPS-stimulated group,both NF-κB activity and concentration of TNF-α were significantly lowered in IL-10＋LPS group.Conclusions LPS might activity NF-κB in the PAMs and induce the increase of transcription and expression of TNF-α gene;IL-10 could inhibit the activation of NF-κB and reduce the release of TNF-α.%目的 探讨白介素-10对内毒素(LPS)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，为临床运用白介素-10提供理论依据。方法 用支气管肺泡灌洗法收集肺泡巨噬细胞(PAM)进行培养，分正常对照组、LPS组、IL-10＋LPS组。用凝胶电泳迁移率改变分析(EMSA)法和ELISA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 LPS组NF-κB活性和TNF-α含量在刺激后0.5～4h明显高于正常对照组；IL-10＋LPS组NF-κB活性和TNF-α含量均显著低于LPS组。结论 LPS诱导PAM的NF-κB活化，导致TNF-α基因表达增强；白介素-10可抑制NF-κB活化而减少TNF-α的释放。