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目的 探讨LPS的直接诱导作用对肺微血管内皮细胞(PMVEC)IL-8表达的影响及通过核因子ΚB(NF-ΚB)的调控机理。方法 以100ng/ml LPS刺激PMVEC 0,0.5,1,2,4,6,8h或1,10,100ng/mlLPS刺激1 h或6 h为检测时相点,ELISA、原位杂交试验分别检测的培养液上清中分泌的IL-8及PMVEC内IL-8mRNA的表达;凝胶电泳迁移率分析(EMSA)检测NF-ΚB的活化;并观察NF-ΚB活化抑制对IL-8表达的影响。结果 LPS能显著促进PMVEC表达IL-8,包括促进IL-8mRNA的表达及IL-8的分泌,在时间上mRNA的表达先于IL-8分泌;且LPS的直接诱导能迅速活化NF-KB,1h达到高峰,后逐渐下降。PDTC能显著抑制NF-KB的活化及IL-8的表达(P<0.01)。结论 表明细菌致病因子LPS的直接诱导确能通过促进NF-B的活化,从而启动IL-8的高效表达和分泌,为多形核中性粒细胞(PMN)的迁移提供必需的物质条件,导致肺损伤。
Objective To investigate the effect of direct induction of LPS on the expression of IL-8 in pulmonary microvascular endothelial cells (PMVEC) and the regulation of nuclear factor ΚB (NF-ΚB). Methods The cultures were incubated with LPS at 100ng / ml for 0, 0.5, 1, 2, 4, 6, 8h or 1, 10 and 100ng / ml LPS stimulation for 1 h or 6 h for detection, ELISA and in situ hybridization The expression of IL-8 and IL-8 mRNA in PMVEC secreted in the supernatant of the liquid were measured. The activation of NF-κB was detected by electrophoretic mobility shift assay (EMSA). The effect of NF-κB activation on IL-8 expression was also observed. Results LPS could significantly promote the expression of IL-8 in PMVEC, including promoting the expression of IL-8 mRNA and the secretion of IL-8. The mRNA expression of IL-8 was earlier than the secretion of IL-8. LPS direct induction of NF- 1h reached its peak, then gradually decreased. PDTC can significantly inhibit the activation of NF-KB and the expression of IL-8 (P <0.01). The results showed that the direct induction of bacterial pathogenic factor LPS can indeed activate NF-B, and then initiate the high expression and secretion of IL-8, providing the necessary material conditions for the migration of polymorphonuclear neutrophils (PMN) Lead to lung damage.