Antioxidative treatment inhibits the release of thrombogenic tissue factor from irradiation- and cytokine-induced endothelial cells
Antioxidative treatment inhibits the release of thrombogenic tissue factor from irradiation- and cytokine-induced endothelial cells
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DOI：
10.1016/j.cardiores.2006.12.018
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发表时间：
2007-03-01
影响因子：
10.8
通讯作者：
Rauch, Ursula
中科院分区：
文献类型：
--
作者：
Szotowski, Bjoern;Antoniak, Silvio;Rauch, Ursula
目的：研究抗氧化剂吡咯烷二硫代氨基甲酸酯(PDTC)和N-乙酰半胱氨酸(NAC)对电离辐射(IR)和肿瘤坏死因子-α(TNF-α)诱导的人脐静脉内皮细胞(HUVECs)组织因子(TF)表达和释放的影响。进行实时定量聚合酶链式反应、酶联免疫吸附试验和转铁蛋白活性检测，包括培养上清液中转铁蛋白活性。通过流式细胞仪检测活性caspase-3的表达和化学发光法检测细胞内活性氧自由基(ROS)的形成。结果：IR和TNF-α处理HUVEC后，Tf的表达持续存在，至今尚未见报道。IR和TNF-α联合刺激导致微粒子相关因子大量脱落，并与细胞凋亡和ROS形成呈正相关。抗氧化前处理不仅减少了细胞的凋亡和ROS的形成，而且还减少了血栓形成微粒的释放。结论：抗氧化处理抑制了微粒的凋亡和脱落，从而降低了血栓形成能力。因此，当抗氧化剂与抗凝剂联合使用时，可能有助于防止抗增殖治疗后的晚期血栓形成。(C)2006年欧洲心脏病学会。爱思唯尔出版，版权所有。
Objectives: The aim of this study was to investigate the effect of the antioxidants pyrrolidine dithiocarbamate (PDTC) and N-acetylcysteine (NAC) on the ionizing radiation (IR)- and tumor necrosis factor-alpha (TNF-alpha) induced tissue factor (TF) expression and its release from human umbilical vein endothelial cells (HUVECs).Methods: HUVECs were irradiated with a single dose of either 5 Gy or 10 Gy and stimulated with TNF-alpha (10 ng/mL) in the presence or absence of PDTC and NAC, respectively. Quantitative real-time PCR, ELISA, and TF activity measurements were performed, including TF activity in the supernatant. Apoptosis was detected by flow cytometric active caspase-3 measurement and formation of reactive oxygen species (ROS) by chemiluminescence.Results: We demonstrated a thus far uninvestigated persistent induction of TF expression in HUVECs after treatment with IR and TNF-alpha. Combined stimulation with IR and TNF-alpha led to an immense shedding of microparticle-associated TF which was positively correlated with apoptosis and ROS formation. Antioxidative pre-treatment reduced not only apoptosis and ROS formation, but also the release of thrombogenic microparticles.Conclusions: Antioxidative treatment inhibited apoptosis and shedding of microparticles, thereby reducing thrombogenicity. Thus, antioxidants may help to prevent late thrombosis after antiproliferative treatment when used in combination with anticoagulants. (c) 2006 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.