摘要
目的:探讨胰蛋白酶对人上皮细胞白细胞介素-8(IL-8)分泌的影响.方法:人肺上皮细胞系A549细胞分别接种于12孔培养板各孔内,并分别用不同浓度的胰蛋白酶和/或胰蛋白酶抑制剂进行刺激.刺激时间为2h、8h和16h.用ELISA方法检测上清液中的IL-8水平.结果:经过16h的培养,胰蛋白酶可引起浓度相关性IL-8释放,胰蛋白酶在浓度1μg/L时就可引起IL-8的释放量增加,3μg/L时诱导IL-8的释放量达高峰,为基础分泌量的5倍,但随着胰蛋白酶浓度的增加,IL-8的释放量反而下降.胰蛋白酶抑制剂可以抑制胰蛋白酶诱导IL-8释放的作用.时间相关曲线表明,胰蛋白酶从2h起即可引起IL-8释放,16h达高峰.结论:胰蛋白酶可促进人肺上皮细胞分泌IL-8,表明它能积极参与呼吸道的炎症过程.
AIM： To investigate the actions of trypsin on the secretion of interleukin-8 （IL-8） from human lung epithelial cells. METHODS： A549 cells were cultured in a 12well culture plate. The challenge was performed by adding various concentrations of trypsin or trypsin inhibitor into each well, respectively. After 2 h, 8 h or 16 h, the reactions were terminated by removing the supernatant from each well. A sandwich ELISA was used to determine the levels of IL-8 in supernatants. RESULTS： Following 16 h incubation, trypsin induced the secretion of IL-8 in a concentration dependent manner. As low as 1μg/L trypsin was able to induce IL-8 release from epithelial cells and the maximum of accumulated release of IL-8 was observed with 3μg/L trypsin, which was 5-fold more than the baseline release. However, when trypsin concentration increased over 3μg/L, the extent of increased secretion of IL-8 decreased. Soybean trypsin inhibitor （SBTI） and αl-antitrypsin （α1-AT） inhibited trypsin induced secretion of IL-8. The time course showed that the actions of trypsin initiated at 2 h and reached their peak at 16 h. CONCLUSION： Trypsin is a potent secretogogue of IL-8 release from cultured human lung epithelial cells, indicating that it is likely to be involved in the pathophysiological process of airway inflammation.
出处
《第四军医大学学报》
CAS
北大核心
2005年第16期1445-1447,共3页
Journal of the Fourth Military Medical University
基金
广东省科技计划重点项目(2003B31502)
李嘉诚基金资助项目(C0200001)
