旋覆花内酯抑制炎性因子介导的内皮细胞与单核细胞的黏附

Acetylbritannilactone Inhibits Inflammatory Stimulus-mediated Adhesion between Endothelial Cells and Macrophages

为了观察旋覆花内酯(1-O-acetylbritannilactone,ABL)乙酰化衍生物ABLO2对脂多糖(lipopolysaccharide,LPS)/干扰素-γ(interferon-γ,IFN-γ)刺激的内皮细胞ECV304活化及其与RAW264.7单核/巨噬细胞相互作用的影响,采用Western印迹检测核因子-κB(nuclearfactor-κB,NF-κB)活化以及NF-κB依赖的黏附分子的表达水平,应用电泳迁移率改变分析(electrophoretic mobility shift assay,EMSA)检查ABLO2预处理及LPS/IFN-γ诱导对NF-κB与DNA结合活性的影响。结果显示,ABLO2显著抑制LPS/IFN-γ诱导的NF-κB核转位和DNA结合活性,同时ABLO2降低NF-κB抑制蛋白(IκB)激酶(IκBkinases,IKK)的活性,抑制IκB的磷酸化及降解;ABLO2还通过减少血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)、骨桥蛋白(osteopontin,OPN)、基质金属蛋白酶-9(metalloproteinase-9,MMP-9)、黏蛋白-c(tenascin-c)的表达,进而减弱单核细胞与内皮细胞之间的黏附作用。研究结果表明,ABLO2通过抑制IKK活性及IκB降解而抑制NF-κB活化,进而起到抑制NF-κB依赖的黏附分子表达及细胞黏附作用。

To investigate the effects of 1,6-O2-diacetylbritannilactone （ABLO2） on the activation of endothelial cell ECV304 and its interaction with macrophages treated with lipopolysaccharide （LPS）/interferon- （IFN-γ）. Western blot analysis was adopted to measure the nuclear translocation of nuclear factor-κb （NF-κB） and the expression of some adhesion molecules. Electrophoretic mobility shift assay （EMSA） was used to detect DNA- binding activity of NF-κB in ECV304 cells pretreated with ABLO2. The results showed that ABLO2 inhibited the NF- κB activation by blocking Ir, B kinase （IKK） activation, and suppressing inhibitor of NF-κB （I-κB） phosphorylation and degradation. In addition, ABLO2 could inhibit the adhesion between endothelial cells and macrophages by decreasing the expression of vascular cell adhesion molecule-1 （VCAM-1）, osteopontin （OPN）, matrix metalloproteinase-9 （MMP-9） and tenascin-c. These results suggest that ABLO2 is a potent agent to prevent vascular inflammatory disease in the vessel.