凝血酶引起人支气管上皮细胞内活性氧及血小板源生长因子-AB分泌的改变  

Changes of reactive oxygen species and platelet-derived growth factor-AB caused by thrombin in human bronchial epithelial cells

作　　者：曹守冬 肖伟[2] 

机构地区：[1]煤炭总医院急诊科,北京100028 [2]山东大学齐鲁医院呼吸科,济南250012

出　　处：《中华哮喘杂志（电子版）》2011年第3期19-22,共4页Chinese Journal of Asthma(Electronic Version)

摘　　要：目的探讨凝血酶对永生化人支气管上皮细胞(BEP2D细胞)内活性氧产生及血小板源生长因子-AB(PDGF-AB)分泌的影响。方法不同浓度凝血酶刺激指数生长期的BEP2D细胞,通过检测氧化型氢化乙啶及二氯荧光素荧光强度测定活性氧簇含量变化。采用双抗体夹心ELISA法检测BEP2D细胞分泌PDGF-AB的变化。结果随着凝血酶浓度增加,反应体系中活性氧明显升高,且有明显的剂量反应关系。凝血酶处理组BEP2D细胞上清液中PDGF-AB含量较对照组上清液显著升高[(770.33+24.29)ng/L vs(117.42±10.85)ng/L,P<0.01]。凝血酶在一定范围内有剂量反应关系。10 U/ml凝血酶刺激48 h后BEP2D细胞分泌PDGF-AB量最大[(817.63+22.53)ng/L]。结论凝血酶既可以引起细胞内氧化应激导致基因毒性,又可以通过刺激BEP2D细胞分泌PDGF-AB发挥自分泌及旁分泌作用。Objective To investigate the effects of thrombin on reactive oxygen species and platelet-derived growth factor-AB（PDGF-AB） in immortalized human bronchial epithelial cells（BEP2D cells）.Methods BEP2D cells at exponential growth phase were stimulated by different concentrations of thrombin.The content of active oxygen species was determined by detecting the fluorescence intensity of hydroxyethidium and dichlorofluorescein.The changes of PDGF-AB were measured by double antibody sandwich ELISA assay.Results With the increase of the thrombin concentration,reactive oxygen species significantly increased,and there was a dose-response relationship.The concentration of PDGF-AB in culture supernatants in thrombin-treated group was higher than that in control group [（770.33＋24.29）ng/L vs（117.42＋10.85） ng/L,P〈0.01],and there was a dose-response relationship.The concentration of PDGF-AB was maximal [（817.63＋22.53） ng/L] when BEP2D cells were stimulated with 10 U/ml thrombin for 48 hours.Conclusions Thrombin can induce cellular oxidative stress to lead to genotoxicity,and play autocrine and paracrine role by stimulating the secretion of PDGF-AB in BEP2D cells.

关 键 词：凝血酶 永生化人支气管上皮细胞 活性氧簇 血小板源生长因子-AB 

分 类 号：R99[医药卫生—毒理学]