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目的 探讨脂多糖 (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 10 0ng/mlLPS刺激PMVEC 0 ,0 .5 ,1,2 ,4,6,8h或 1ng/ml、10ng/ml、10 0ng/mlLPS刺激 1h或6h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)检验NF κΒ的活化 ;并观察NF κΒ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 κΒ ,1h达到高峰 ,后逐渐下降。PDTC能显著抑制NF κΒ的活化及IL 8的表达 (P <0 .0 1)。结论 表明细菌致病因子LPS的直接诱导确能通过促进NF κΒ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 lipopolysaccharide (LPS) on the expression of IL-8 in pulmonary microvascular endothelial cells (PMVEC) and its regulation by NF-κB. Methods PMVEC was stimulated with 10 0 ng / ml LPS for 0, 0.5, 1, 2, 4, 6, 8 h or 1 ng / ml, 10 ng / ml and 10 0 ng / ml LPS for 1 h or 6 h. The expression of IL-8 mRNA in IL-8 and PMVEC secreted in the culture supernatant was detected by hybridization assay. 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. MRNA expression over time precedes secretion of IL8. And direct induction of LPS can rapidly activate NF κΒ, peaked at 1h, and then decreased gradually. PDTC significantly inhibited NF-κB activation and IL-8 expression (P <0.01). The results showed that the direct induction of bacterial pathogenic factor LPS can indeed promote the activation of NF-κB, and then initiate the high expression and secretion of IL-8, providing the necessary material conditions for the migration of polymorphonuclear neutrophils (PMNs) damage.