LPS的直接诱导对肺微血管内皮细胞内NF-κB的影响

Effect of the direct induction of LPS on the nuclear factor-κappa B in pulmonary micro vascular endothelial cells

目的 :探讨LPS的直接诱导作用对肺微血管内皮细胞 (PMVEC)核因子kB (NF -κB)的影响。方法 :10 0ng/mlLPS刺激PMVEC 0h、 0 5h、 1h、 2h、 4h、 6h、 8h或 10ng/ml、 5 0ng/ml、 10 0ng/mlLPS刺激 1h,凝胶电泳迁移率试验检测NF -κB的活化 ,免疫细胞化学观察其亚基p5 0、p6 5的核转位。并通过加入活化阻断剂TPCK观察对诱导的NF -κB活化的影响。结果 :LPS的刺激迅速诱导p5 0、p6 5亚基核转位 ,活化NF -κB ,1h达到高峰 ,且呈剂量依赖关系 ,后逐渐下降。PDTC能显著抑制其活化 ,P <0 0 1。结论 :LPS的直接刺激诱导NF -κB的活化 。

Objective:To study the effect of the direct induction of LPS on the nuclear factor-κappa B(NF-κ B)in pulmonary micro vascular endothelial cells(PMVEC).Methods:LPS with the concentration of 100ng/ml was used to stimulate PMVEC for 0h?0 5h?1h?2h?4h?6h?8h or LPS was used to stimulate PMVED for 1h at different concentration of 10ng/ml,50ng/ml and 100ng/ml.Electrophoretic mobility shift assay(EMSA)was employed to detect the activation of NF-κ B and immunocytochemistry was used to observe the nuclear translocation of two subunits p50 and 65.Activation blocking agent TPCK was used to observe its effect on the activation of NF-κ B.Results:LPS stimulation can quickly induce the nuclear translocation of p50 and p65 and activate NF-κ B which peaked at 1h in dose-dependent manner and then decreased.PDTC can significantly inhibit the activation of NF-κ B induced by LPS(P<0 01).Conclusion:The direct simulation of LPS can induce the activation of NF-κ B,which might play an important role in the inflammatory response induced by LPS.