水蛭素对凝血酶诱导血管内皮细胞与中性粒细胞表达黏附分子的影响

Effects of hirudin for the thrombin-induced expression of adhesive molecules in neutrophils and human umbilical vein endothelial cells

目的:研究水蛭素对凝血酶诱导人脐静脉内皮细胞(HUVEC)表达细胞黏附分子(ICAM-1)和中性粒细胞表达膜整合素CD11b的影响。方法:用酶消化法获得HUVEC,密度梯度离心和重力沉降法分离人外周血细胞中性粒细胞。用酶联免疫吸附测定HUVEC在凝血酶、水蛭素作用下ICAM-1的变化,同时以免疫荧光流式细胞术检测中性粒细胞表达CD11b水平的变化。结果:与对照组相比,凝血酶诱导的HUVEC表达ICAM-1显著增多,随时间延长,ICAM-1的表达递增,高峰在24h。凝血酶使中性粒细胞CD11b的表达也显著上调,6h平均荧光强度明显增加,随时间延长逐渐升高,高峰在24h。水蛭素和肝素对凝血酶诱导的HUVEC表达ICAM-1和中性粒细胞表达CD11b均有明显的抑制作用,且水蛭素抑制ICAM-1的表达强于肝素,尤其在ICAM-1表达的高峰期,对CD11b表达的抑制作用水蛭素与肝素无明显差别。结论:水蛭素可以明显抑制凝血酶诱导的HUVEC表达ICAM-1和中性粒细胞表达CD11b。

Objective： To investigate effects of hirudin for the thrombin-induced expression of intercellular adhesive molecules-1 （ICAM-1 ） in human umbilical vein endothelial cells （HUVEC） and cluster of differentiation （CD） lib in neutrophil granucytes. Methods： The HUVEC was purified by digestion with collagenase type I. An enzyme linked immunosorbent assay was used to analyze the expression of ICAM-1 thrombin promoted in the HUVEC. The neutrophil granucytes were purified by density gradient centrifugation and gravimetric sedimentation. An immunofluorence flow cytometery was utilized to observe the expression of CD11b thrombin induced in the neutrophil granucytes. Hirudin was incubated with the HUVEC and neutrophil granucytes, and the changes of ICAM-1 and CDllb were monitored, respectively. Results： Thrombin significantly increased both CD11b and ICAM-1 levels with the maximal peak values at 24 hours, which were positively proportional to the time thrombin exposed to the HUVEC and neutrophil granucytes. Hirudin inhibited the expression of ICAM-1 in the HUVEC more greatly than heparin, and CD11b in the neutrophil granucytes the same as heparin. Conclusion： Hirudin inhibited the generation of both ICAM-1 and CD11b by altering leukocyte adhesion and migration, which suggested affecting the atherosclerotic process.