海水对肺腺癌细胞株A549细胞蛋白酶激活受体2表达的作用研究  

The effect of seawater on expression of protease-activated receptor 2 in adenocarcinoma of lung cell line A549 cells

作　　者：邓朝霞[1] 钱桂生[1] 杨昱[1] 周长喜[1] 王关嵩[1] 

机构地区：[1]第三军医大学新桥医院全军呼吸内科研究所,重庆400038

出　　处：《中国危重病急救医学》2009年第11期648-651,共4页Chinese Critical Care Medicine

基　　金：国家自然科学基金项目（30770928）;全军“十一五”计划专项课题（06G083）;重庆市自然科学基金项目（2007BB5072）

摘　　要：目的观察海水对肺腺癌细胞株A549细胞的炎性损伤及对蛋白酶激活受体2（PAR-2）表达的影响，探讨海水导致急性肺损伤的作用机制。方法将培养的A549细胞接种于6孔板中，按随机数字表法把细胞分为对照组及海水处理2、4、8h组，相应时间点用实时荧光定量逆转录-聚合酶链反应（RT—PCR）和蛋白质免疫印迹法（Western blotting）检测PAR-2的mRNA和蛋白表达；收集细胞培养上清液，用酶联免疫吸附法（ELISA）检测肿瘤坏死因子-α（TNF—α）和白细胞介素-8（IL-8）含量。结果用海水处理后，A549细胞PAR-2 mRNA和蛋白表达均于2h显著增加（P均〈0．05），4h达高峰，分别为对照组的1．8倍和2．2倍，随后下降，但仍显著高于对照组（P均〈0．01）；TNF—α和IL-8含量则于2h即显著升高并达最高峰[分别为（214．35±20．85）ng／L，（55.86±5．65）ng／L]，随后下降，但仍显著高于对照组[分别为（25．86±3．85）ng／L，（6．97±1．77）ng／L，P均〈0．01]。结论海水可致A549细胞炎症反应明显，并可诱导A549细胞PAR-2表达增高。Objective To investigate the effect of seawater on expression of protease-activated receptor 2 （PAR-2） in adenocarcinoma of lung cell line A549 cells, and the inflammatory injury on A549 cells induced by seawater. Methods A549 cells were randomly divided into four groups： control group, 2 hours group, 4 hours group, 8 hours group, in which cells were treated with seawater for 2, 4 and 8 hours respectively. After seawater treatment, cells were collected for real-time quantitative reverse transcrition- polymerase chain reaction （RT-PCR） and Western blotting analysis to determine the expression of PAR-2 mRNA and its protein. Supernatant were collected for tumor necrosis factor-α （TNF-α） and interleukin-8 （IL-8） which were determined by enzyme-linked immunosobent assy （ELISA）. Results The expression of PAR-2 mRNA and protein of A549 cells increased in a time-dependent manner after seawater treatment, significantly so after 2 hours in all groups （both P〈0. 05）, and peaked at 4 hours after seawater treatment （1.8-fold and 2.2-fold respectively, both P〈0.01）, followed by a decrease though still higher than those of control group significantly （both P〈0. 01）. TNF-α and IL-8 in supernatant increased significantly after seawater treatment, peaking at 2 hours after seawater treatment （（214.35±20.85） ng/L, （55.86 ± 5.65） ng/L] and then followed by a slight decrease though still significantly higher than those of control group [（25.86±3.85） ng/L, （6.97±1.77） ng/L, all P〈0.01]. Conclusion Seawater can induce significant inflammation of A549 cells and up-regulate the expression of PAR-2 on A549 cells.

关 键 词：海水 蛋白酶激活受体2 炎症 

分 类 号：R686[医药卫生—骨科学]