论文部分内容阅读
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TNF α表达可能还与其他核转录因子有关
To investigate the effect of endotoxin stimulation on the activity of nuclear transcription factor AP 1 in rat alveolar macrophages (AM), and to explore the role of AP 1 in the regulation of TNFα production by alveolar macrophages stimulated by LPS in rats, The dynamic changes of DNA-binding activity of AP 1 in AM nucleoprotein of rats stimulated by LPS (Ecoli0 2 6: B6) were detected by EMSA. Transgenic technique was used to transfect the AP1 thio-oligonucleotide S ODNdecoy was transplanted into AM and the effect of AP 1 thio-oligonucleotide traps on the expression of TNF α in AMs stimulated by LPS was observed by ELISA. The results showed that the AP 1 was rapidly activated and peaked at 10 h after administration of 10 ng / ml LPS. The activity of AP 1 decreased slightly at 1 h and then gradually rose at 3 h. The activity of AP 1 decreased rapidly at 5 h and 8 h. At least for 8h, and showed a dose-effect relationship with LPS stimulation. The S ODN trap of AP 1 could significantly inhibit but not completely block LPS-induced TNFα expression. It indicates that LPS can activate AP1 in rat AM, which may play an important role in LPS-induced inflammation. AP1 may play an important regulatory role in LPS-induced TNFa expression in AM, but TNFα Expression may also be related to other nuclear transcription factors