白黎芦醇对fMLP诱导中性白细胞与人脐静脉内皮细胞粘附的抑制作用及机制

The inhibitive effects of resveratrol on adhesion of neutrophils to HUVECs induced by fMLP

目的研究白藜芦醇（resveratrol,Res）对趋化肽N-甲酰-甲硫氨酰-亮氨酰苯丙氨酸（fMLP）诱导中性白细胞与人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVECs）的粘附及HUVECs释放可溶性粘附分子的影响。方法观察不同浓度Res对fMLP（10μmol·L^-1）刺激HU-VECs与中性白细胞粘附的影响。以ELISA法测定可溶性细胞间粘附分子,血管细胞粘附分子和E-选择素。以Fura-3和海萤萤光素诱导剂为荧光指示剂分别测定中性白细胞内[Ca^2＋]i水平和O2^-·浓度。结果Res（1-50μmol·L^-1）明显抑制fMLP诱导的中性白细胞与HUVECs的粘附反应,同时,明显抑制fMLP诱导粘附分子的释放效应。Res抑制中性白细胞中O2^-·生成和[Ca^2＋]i升高,其中Res5μmol·L^-1和50μmol·L^-1与fMLP诱导组相比具有统计学意义（P〈0.01）。结论白藜芦醇通过影响中性白细胞内O2^-·生成和[Ca^2＋]i水平,抑制中性白细胞与内皮细胞的粘附以及细胞间粘附分子表达,在阻断炎性反应的发生与发展过程中发挥着重要作用。

Aim To study the effects of resveratrol （Res） on the release of soluble adhesion molecules from human umbilical vein endothelial cells （HUVECs） induced by N-formyl-methionyl-leucyl-phenylalanine （fMLP） and the adhesion between neutrophils and HUVECs. Methods The effects of Res on neutrophils adhesion to human umbilical endothelial cell （HUVECs）triggered by fMLP were examined. The soluble intercellular cell adhesive molecule-1 （ ICAM-1 ） , the soluble vascular cell adhesive molecule-1 （VCAM-1） and E-selectin release from fMLP （ 10 μmol·L^-1 ） stimulated HUVECs were determined by ELISA kits. The neutrophils isolated from human vein blood were loaded with Fluo-3, a fluorescent indicator, to detect intracellular free calcium concentration （ [ Ca^2＋] i ） , and CLA was used as a chemiluminescent indicator to determine superoxide production in neutrophils. Results Res（ 1 -50 μmol·L^-1） significantly inhibited neutrophil adhesion to fMLP-stimulated HUVECs and also obviously downregulated the levels of ICAM-1, VCAM- 1 and E-selectin in supernatant of HUVECs culture stimulated by fMLP in the dose-dependent pattern. Res also suppressed fMLP-activated superoxide generation and [ Ca^2＋ ]i increase in neutrophils. Conclusions Res involved in neutrophil adhesion to HUVECs intermediated by cell adhesive molecules expression trigged by [ Ca^2＋ ]i and superoxide production in neutrophils, which means a lot to prevent inflammatory diseases.