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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)的调控机制。方法以100ng/ml LPS刺激PMVEC 0、0.5、1、2、4、6、8h或1ng/ml、10ng/ml1、00ng/ml LPS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-κB,1h达到高峰,后逐渐下降。PDTC能显著抑制NF-κB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-κB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 effects of lipopolysaccharide (LPS) on the expression of IL-8 in pulmonary microvascular endothelial cells (PMVEC) and the regulation of NF-κB by direct induction of lipopolysaccharide (LPS). Methods PMVEC was stimulated with 0, 0.5, 1, 2, 4, 6, 8h or 1ng / ml of LPS at 100ng / ml for 1h or 6h with 10ng / ml LPS for 1h and 6h. ELISA, in situ hybridization The levels of IL-8 and IL-8 mRNA in PMVEC secreted by supernatant of culture supernatant were detected respectively. The activation of NF-κB was detected by electrophoretic mobility shift assay (EMSA). The effect of NF-κB activation on the expression of IL-8 Impact. Results LPS can 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 IL-8 secretion. The direct induction of LPS can rapidly activate NF-κB, peaked at 1h and then decreased gradually. PDTC can significantly inhibit the activation of NF-κB 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-κB, and then initiate the high expression and secretion of IL-8, providing the necessary material conditions for the migration of polymorphonuclear neutrophils (PMNs) Lead to lung damage.