LPS的直接诱导对肺微血管内皮细胞IL-8的表达及对PMN趋化作用影响的研究

目的探讨LPS的直接诱导作用对肺微血管内皮细胞(PMVEC)IL -8表达的影响及诱导发生的PMVEC对多形核中性粒细胞 (PMN)趋化作用的影响。方法100ng/mlLPS刺激PMVEC0、1、2、4、6、8h或1、10、100ng/mlLPS刺激6h,ELISA和原位杂交试验分别检测培养液上清中分泌的IL -8及PMVEC内IL-8mRNA的表达 ,同时琼脂糖平板法检测对PMN的趋化作用 ,并通过抗IL -8的抗体抑制试验观察对趋化作用的影响。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。LPS能促进PMVEC对PMN的趋化 ,随刺激作用的持续 ,诱导的PMVEC对PMN的趋化作用加强。抗IL -8抗体能显著抑制对PMN的趋化作用(P<0.01)。结论表明细菌致病因子LPS的直接诱导确能促进PMVEC高效表达和分泌IL -8 ,从而为PMN的迁移提供必需的物质条件,发挥对PMN的趋化作用而参与导致肺损伤。

An Research on Expression of IL-8 and Attraction to PMN of PMVEC Directly- Induced by LPS

Objective To study the changes of pulmonary microvascular endothelial cells'(PMVEC)ex-pression of IL-8and its attraction to PMN directly induced by LPS.Methods With methods of ISH,ELISA re-spectively to examine the expressions of IL-8mRNA in PMVEC and its protein secretion in the supernatant of culture medium,when PMVEC is directly excited by100ng/ml LPS at0h,1h,2h,4h,6h,8h or1ng/ml,10ng/ml,100ng/ml LPS at6h.At the same time,to examine the attraction index of PMVEC to PMN with the agarose plate assay and the inhibited effects on it by using anti-IL-8antibody.Results LPS could directly act on the PMVEC to promote the IL-8expression,including promoting the expression of IL-8mRNA and secretion of IL-8pro-tein,and the mRNA expressions increased before the protein secreted in terms of time.The LPS excitement could also significantly promote the attraction of PMVEC to PMN,while anti-IL-8antibody could significantly restrain this attraction.Conclusions The directly-induction of LPS could promote PMVEC to the expression and secretion of IL-8.So providing the required physical condition for the PMN migration,and then intensifying the attraction to PMN.It could play an important role in inflammatory response induced by LPS.