核因子-κB在内毒素刺激大鼠肺泡巨噬细胞中的动态变化及其在肿瘤坏死因子-α表达中的作用  

Kinetics of nuclear factor κB in LPS stimulated rat alveolar macrophages and its regulative effect on tumor necrosis factor α expression

作　　者：李春穴[1] 蒋建新[1] 姜燕平 单佑安[1] 冯凯[1] 张良[1] 

机构地区：[1]400042,重庆,第三军医大学附属大坪医院野战外科研究所第四研究室 [2]解放军第三零七医院耳鼻喉科

出　　处：《中华创伤杂志》2003年第5期308-311,共4页Chinese Journal of Trauma

基　　金：国家重点基础研究发展规划资助项目(G19990 5 42 0 3);高等学校全国优秀博士学位论文作者专项资金资助项目;教育部优秀年轻教师基金资助项目

摘　　要：目的　观察内毒素 (LPS)刺激对大鼠肺泡巨噬细胞 (alveolarmacrophages,AM)中核因子 -κB(NF -κB)活性的影响 ,探讨NF -κB在LPS刺激大鼠AM表达肿瘤坏死因子 -α(TNF -α)中的作用。　方法　体外观察LPS刺激大鼠AM中NF -κB活性的动态变化 ,应用圈套策略观察NF-κB在LPS刺激大鼠AM表达TNF -α中的作用。　结果　LPS刺激可使大鼠AM内NF -κB明显活化 ,并与LPS剂量正相关 ;抗体超迁移率实验结果显示p5 0、p6 5参与了NF -κB的活化 ;NF -κB的圈套硫代寡核苷酸 (S -ODN)能明显抑制LPS刺激大鼠AM表达TNF -α ,但不能完全阻断LPS诱导的TNF -α表达。　结论　NF -κB(p5 0 p6 5 )在LPS介导的炎症反应中发挥重要作用 ,LPS刺激大鼠AM表达TNF -α受NF -κB调控 ,但其他核转录因子可能也在TNF -α的表达中发挥重要作用。Objective To investigate the activation of nuclear factor κB (NF-κB) induced by lipopolysaccharides (LPS) in rat alveolar macrophages (AM) and its regulative role in tumor necrosis factor (TNF-α) expression. Methods The dynamic activity changes of NF-κB DNA induced by LPS (E.coli 026:B6) were determined with electrophoretic mobility shift assay (EMSA). The phosphorothioate oligodeoxynucleotides (S-ODN) decoy was transfected into AM 12 hours prior to LPS stimulation. The effect of NF-κB S-ODN decoy on expression of TNF-α in AM stimulated by LPS were measured with enzyme-linked immunosorbent assay (ELISA) kit. Results NF-κB could be activated remarkably after 0.5 hour of LPS stimulation at concentration of 100 ng/ml, reached the highest level 1 hour after LPS stimulation and gradually decreased. But the activation of NF-κB could last at least 8 hours. The dose for LPS stimulation was related to activation of NF-κB in a dose-dependent fashion, ie, the activation of NF-κB gradually strengthened with dose increase of LPS. Supershift assays proved that p50 and p65 were involved in the activation of NF-κB. NF-κB S-ODN decoy could markedly (not completely) inhibit LPS-induced TNF-α production. Conclusions NF-κB plays an important role in LPS induced inflammatory response. However, entire inhibition of the activity of NF-κB can not completely prevent TNF-α expression induced by LPS in rat AM, which implies that other nuclear factors may participate in TNF-α expression.

关 键 词：核因子-ΚB 内毒素 炎症 巨噬细胞 肿瘤坏死因子-Α 圈套硫代寡核苷酸 

分 类 号：R363[医药卫生—病理学]