Exogenous superoxide mediates pro-oxidative, proinflammatory, and procoagulatory changes in primary endothelial cell cultures
Exogenous superoxide mediates pro-oxidative, proinflammatory, and procoagulatory changes in primary endothelial cell cultures
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DOI：
10.1016/j.freeradbiomed.2005.06.010
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发表时间：
2005-11-01
影响因子：
7.4
通讯作者：
Sela, S
中科院分区：
文献类型：
--
作者：
Jacobi, J;Kristal, B;Sela, S
内皮功能障碍/活化是长期心血管并发症和动脉粥样硬化发展的基础。本研究的目的是研究外源性亚致死流量的超氧化物对内皮细胞功能障碍的直接作用。人脐静脉内皮细胞（HUVEC）暴露于由0.1 mM黄嘌呤和4 mU/ml黄嘌呤氧化酶产生的超氧化物15分钟，并检查基本的内皮功能。超氧化物歧化酶和/或过氧化氢酶被用作O-2（中心点-）/H2 O2的清除剂，以确定关键的罪魁祸首。HUVEC的脱落通过中性红摄取测定，凋亡通过膜联蛋白V结合测定。通过IL-8 mRNA表达和细胞粘附分子（CAM）来评估炎症。eNOS和iNOS信息和eNOS蛋白作为NO的间接测量。通过估计细胞内组织因子来评估凝血状态。光泽精化学发光法测定内皮细胞NADPH氧化酶活性。诱发的亚致死超氧化物剂量：（1）促炎状态，表现为内皮表面IL-8 mRNA表达和CAM增加，（2）HUVEC凋亡和激活内皮NADPH氧化酶，（3）细胞内组织因子增加，（4）eNOS mRNA和蛋白减少和iNOS mRNA上调。我们的结论是，细胞外低通量的超氧化物具有多效性的特点，触发激活/功能障碍的内皮细胞。(c)2005年爱思唯尔公司All rights reserved.
Endothelial dysfunction/activation underlies the development of long-term cardiovascular complications and atherosclerosis. The aim of this study was to examine a direct role for exogenous sublethal flux of superoxide on endothelial cell dysfunction. Human umbilical vein endothelial cells (HUVEC) were exposed to superoxide generated by 0.1 mM xanthine and 4 mU/ml xanthine oxidase for 15 min and essential endothelial functions were examined. Superoxide dismutase and/or catalase was used as scavenger for O-2(center dot-)/H2O2 to determine the key culprit. HUVEC detachment was determined by neutral red uptake and apoptosis by annexin V binding. Inflammation was estimated by IL-8 mRNA expression and cellular adhesion molecules (CAM). eNOS and iNOS message and eNOS protein served as an indirect measure for NO. Procoagulable state was evaluated by estimating the intracellular tissue factor. Activation of endothelial NADPH oxidase was determined by lucigenin chemiluminescence. Sublethal superoxide dose evoked: (1) proinflammatory state manifested by increased IL-8 mRNA expression and CAM on the endothelial surface, (2) HUVEC apoptosis and activated endothelial NADPH oxidase, (3) increase in intracellular tissue factor, and (4) decrease in eNOS mRNA and protein and up-regulation of iNOS mRNA. We conclude that extracellular low flux of superoxide exhibits pleiotropic characteristics, triggering activation/dysfunction of endothelial cells. (c) 2005 Elsevier Inc. All rights reserved.