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目的:研究胰蛋白酶对IL-8释放的影响。方法:分离、培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)、倒置显微镜观察形态变化,流式细胞术检测内皮细胞标志和蛋白酶活化受体-2(proteinase-activated receptor-2,PAR-2)表达,ELISA检测HUVECs培养上清中IL-8水平。结果:HUVECs表达内皮细胞标志和PAR-2。刺激16 h,1 g/ml胰蛋白酶和100M PAR-2激活肽组HUVECs单层均匀性降低。胰蛋白酶能够显著刺激HUVECs释放IL-8,PAR-2激活肽也诱导IL-8水平升高。蛋白酶抑制剂和PAR-2抑制肽均能够显著抑制胰蛋白酶诱导的IL-8释放。PAR-2激活肽和胰蛋白酶诱导升高的IL-8水平之间成正相关性。结论:胰蛋白酶很可能通过PAR-2激活促进血管内皮细胞释放IL-8。
Objective: To study the effect of trypsin on the release of IL-8. Methods: Human umbilical vein endothelial cells (HUVECs) were isolated and cultured. The morphological changes were observed under inverted microscope. The expressions of endothelial cell markers and proteinase-activated receptor-2 (PAR) were detected by flow cytometry -2), and the level of IL-8 in the supernatant of HUVECs was detected by ELISA. RESULTS: HUVECs expressed endothelial markers and PAR-2. The monolayer homogeneity of HUVECs was decreased after 16 h stimulation, 1 g / ml trypsin and 100 M PAR-2 activating peptide. Trypsin significantly stimulated the release of IL-8 from HUVECs and PAR-2-activated peptide also induced an increase of IL-8. Both protease inhibitors and PAR-2 inhibitory peptides were able to significantly inhibit trypsin-induced IL-8 release. There is a positive correlation between the PAR-2 activating peptide and trypsin-induced elevated IL-8 levels. Conclusion: Trypsin may promote the release of IL-8 by vascular endothelial cells through PAR-2 activation.