摘要
目的：探讨香烟烟雾提取物（cigarette smoke extract,CSE）诱导脐静脉内皮细胞内质网应激（endoplasmic reticulum stress,ERS）对中性粒细胞趋化作用的影响。方法实验分4组：正常对照组、CSE刺激组、ERS抑制剂组和阴性对照组。培养人脐静脉内皮细胞（human umbilical vein endothelial cell,HUVECs）,给予不同浓度的 CSE 刺激24 h,Real time-PCR 法检测糖调节蛋白78（glucose regulated protein 78,GRP78）、趋化因子8（CXCL-8）、细胞间黏附分子1（intercellular adhesion molecule-1, ICAM-1）、E-selectin mRNA 的表达水平；用 ERS 抑制剂4-苯基丁酸（4-phenylbutyrate,4-PBA）干预,ELISA法检测各组 HUVECs 的 CXCL-8蛋白表达水平,Transwell趋化实验检测各组培养上清对正常人外周血中性粒细胞的趋化作用。结果 CSE 增强 GRP78及CXCL-8、ICAM-1、E-selectin mRNA的表达,并在一定范围内具有浓度依赖性；ERS 抑制剂4-PBA减少CXCL-8的分泌（q=8.370,P〈0.05）,并减弱中性粒细胞的趋化作用（q=4.808,P〈0.05）。结论 CSE诱导 HUVECs ERS,并上调 CXCL-8、ICAM-1、E-selectin的基因转录；4-PBA 减弱中性粒细胞的趋化作用,可能与减少CXCL-8分泌作用有关。
Objective To investigate the effect of endoplasmic reticulum stress （ERS） induced by cigarette smoke extract （CSE） in neutrophil migration.Methods Human umbilical vein endothelial cell （HUVECs） were cultured and divided into 4 groups：normal control group,CSE-stimulated group,CSE-stimulated with 4-PBA groups,CSE-stimulated with negative control groups.HUVECs were cultured and stimulated with CSE at concentrations（0%,5%,10% and 20%）respectively for 24 h,mRNA of CXCL-8,ICAM-1,E-selectin and GRP78 were detected by Real-time PCR.ELISA was performed to test the expression of CXCL-8 protein,and neutrophil migration was detected by Transwell board test.Results The mRNA of CXCL-8,ICAM-1,E-selectin and GRP78 increased in CSE-stimulated HUVECs （P 〈0.05）,furthermore,it was concentration-dependent.4-PBA significantly reduced the expression of CXCL-8 protein （P 〈0.05） and neutrophil migration （P 〈0.05）.Conclusions Cigarette smoke extract induces ER stress in HUVECs,which may play a part in increasing the expressions of CXCL-8,ICAM-1 and E-selectin.The reduction of neutrophil migration may relate to the lower expression of CXCL-8.
出处
《国际呼吸杂志》
2016年第12期936-940,共5页
International Journal of Respiration
