Molecular mechanisms of neutrophil-endothelial cell adhesion induced by redox imbalance
Molecular mechanisms of neutrophil-endothelial cell adhesion induced by redox imbalance
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DOI：
10.1161/01.res.84.5.516
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发表时间：
1999-03-19
影响因子：
20.1
通讯作者：
Aw, TY
中科院分区：
文献类型：
--
作者：
Kokura, S;Wolf, RE;Aw, TY
先前的研究已经暗示了细胞内巯基在核因子-KB的活化和内皮细胞粘附分子的转录调节中的作用。本研究旨在确定内皮细胞谷胱甘肽（GSH）或氧化型谷胱甘肽（GSSG）的变化是否可以改变中性粒细胞的粘附性，并确定GSSG/GSH诱导的粘附反应的分子机制。用0.2mmol/L联胺和1 mmol/L丁硫氨酸亚砜（BSO）处理人脐静脉内皮细胞（HUVEC）6 h，可降低GSH水平，增加GSSG/GSH比值，但无细胞毒性。这些氧化还原变化与缺氧/复氧观察到的变化相似。二酰胺加BSO诱导的巯基/二硫键失衡与中性粒细胞粘附HUVEC的双相增加相关，在15分钟（第1阶段）和240分钟（第2阶段）观察到峰值反应。N-乙酰半胱氨酸处理减弱了两个阶段的中性粒细胞粘附，这表明GSH在粘附反应中的作用。有趣的是，I相粘附与GSH水平呈负相关，但与GSSG/GSH比值无关，而2相中性粒细胞粘附与GSSG/GSH比值呈正相关，但与GSH水平无关。细胞间粘附分子-1和P-选择素特异性单克隆抗体在两个阶段中减弱了增加的中性粒细胞粘附，而抗E-选择素单克隆抗体也减弱了2期反应。用放线菌素D和放线菌酮或含有核因子-κ B或激活蛋白-1同源DNA序列的竞争性双链寡核苷酸预处理显著减弱了2期反应，这涉及从头蛋白合成的作用。HUVECs表面细胞间粘附分子1、P-选择素和E-选择素的表达与1期和2期中性粒细胞粘附反应相关。这项研究表明，内皮细胞GSSG/GSH的变化引起不同内皮细胞粘附分子的转录非依赖性和转录依赖性表面表达，从而导致2相嗜中性粒细胞-内皮细胞粘附反应。
Previous studies have implicated a role for intracellular thiols in the activation of nuclear factor-KB and transcriptional regulation of endothelial cell adhesion molecules. This study was designed to determine whether changes in endothelial cell glutathione (GSH) or oxidized glutathione (GSSG) can alter neutrophil adhesivity and to define the molecular mechanism that underlies this GSSG/GSH-induced adhesion response. Treatment of human umbilical vein endothelial cell (HUVEC) monolayers for 6 hours with 0.2 mmol/L diamide and 1 mmol/L buthionine sulfoximine (BSO) decreased GSH levels and increased the ratio of GSSG to GSH without cell toxicity. These redox changes are similar to those observed with anoxia/reoxygenation. Diamide plus BSO-induced thiol/disulfide imbalance was associated with a biphasic increase in neutrophil adhesion to HUVECs with peak responses observed at 15 minutes (phase 1) and 240 minutes (phase 2). N-Acetylcysteine treatment attenuated neutrophil adhesion in both phases, which indicated a role for GSH in the adhesion responses. Interestingly, phase I adhesion was inversely correlated with GSH levels but not with the GSSG/GSH ratio, whereas phase 2 neutrophil adhesion was positively correlated with GSSG/GSH ratio but not with GSH levels. Intercellular adhesion molecule-1 and P-selectin-specific monoclonal antibodies attenuated the increased neutrophil adhesion during both phases, whereas an anti-E-selectin monoclonal antibody also attenuated the phase 2 response. Pretreatment with actinomycin D and cycloheximide or with competing ds-oligonucleotides that contained nuclear factor-kappa B or activator protein-1 cognate DNA sequences significantly attenuated the phase 2 response, which implicated a role for de novo protein synthesis. Surface expression of intercellular adhesion molecule-1, P-selectin, and E-selectin on HUVECs correlated with the phase 1 and 2 neutrophil adhesion responses. This study demonstrates that changes in endothelial cell GSSG/GSH cause transcription-independent and transcription-dependent surface expression of different endothelial cell adhesion molecules, which leads to a 2-phase neutrophil-endothelial adhesion response.