AP-1在内毒素刺激大鼠肺泡巨噬细胞中的动态变化及其在TNF-α表达中的调控作用  

KINETICS OF AP-1 IN LPS-STIMULATED RAT ALVEOLAR MACROPHAGES AND ITS REGULATION ON TNF-α EXPRESSION

作　　者：李春穴[1] 蒋建新[1] 单佑安[1] 张良[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所,重庆400042

出　　处：《解放军医学杂志》2003年第3期209-211,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家重点基础研究发展规划资助项目 (编号G1 9990 542 0 0 );高等学校全国优秀博士学位论文作者专项资金;教育部优秀年轻教师基金项目资助课题

摘　　要：为观察内毒素刺激对大鼠肺泡巨噬细胞(AM)中核转录因子AP 1活性的影响 ,探讨AP 1在LPS刺激大鼠肺泡巨噬细胞表达TNF α中的基因调控作用 ,应用凝胶迁移阻滞技术(EMSA)检测LPS(Ecoli0 2 6∶B6)刺激大鼠AM核蛋白中AP 1的DNA结合活性的动态变化 ;在LPS刺激前 12h ,应用转基因技术将AP 1的硫代寡核苷酸圈套 (S ODNdecoy)转入大鼠AM ,应用ELISA法观察AP 1硫代寡核苷酸圈套对LPS刺激大鼠AM表达TNF α的影响。结果发现 ,应用 10 0ng/mlLPS刺激大鼠AM 0 5h,AP 1即迅速出现活化并达到峰值 ,在 1h活性略有下降 ,3h活性又逐渐回升 ,5h、8h又迅速回落 ,AP 1的活化状态至少可以持续 8h ,且与LPS刺激呈明显的量效关系 ;AP 1的S ODN圈套能明显抑制但不能完全阻断LPS诱导的TNF α表达。说明LPS刺激可使大鼠AM内AP 1活化 ,其在LPS介导的炎症反应中可能发挥重要作用 ;AP 1在LPS诱导的大鼠AM表达TNF α中可能起重要的调控作用 ,但TNFTo investigate the kinetics of activation of the activator protein-1(AP-1) and elucidate its role in TNF-α expression induced by lipopolysaccharide(LPS) in rat alveolar macrophages(AM), dynamic changes of the activity of AP-1 were detected with electrophoretic mobility shift assay(EMSA). The phosphorothioate oligodeoxynucleotide (S-ODN) decoy was transfected into AM prior to LPS stimulation. The level of TNF-α in culture supernatants was measured with an ELISA kit. The results showed that after LPS stimulation for 0.5 hour, remarkable activation of AP-1 could be detected and reached the highest level. The activation of AP-1 rapidly decreased at 1 hour, then increased at 3 hours again and reduced at 5 and 8 hours after LPS stimulation. The activation of AP-1 could persist at least 8 hours and showed a dose-dependent manner to LPS within 1000ng/ml. AP-1 S-ODN decoys could markedly inhibit the LPS-induced TNF-α production by rat AM, but it could not completely inhibit the production of TNF-α induced by LPS in rat AM. It is suggested that LPS could induce activation of AP-1 in rat AM; AP-1 played an important role in LPS-induced inflammatory response. It is also suggested that AP-1 involved in the regulation on LPS-induced TNF-α production by rat AM, however, entirely inhibition of the activity of AP-1 could not completely prevent TNF-α production by rat AM. It is also implied that other nuclear factors might also play an important role in the regulation on LPS-induced TNF-α expression by rat AM.

关 键 词：转录因子AP-1 内毒素类 肿瘤坏死因子 硫代寡核苷酸圈套 S-ODN 巨噬细胞 肺泡 

分 类 号：R378[医药卫生—病原生物学] R392.11[医药卫生—基础医学]