内质网应激对中性粒细胞趋化作用的影响

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。

The effect of endoplasmic reticulum stress in neutrophil migration

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.