Laboratory of Tissue Immunopharmacology, Department of Internal Diseases and Clinical Pharmacology, Medical University of Lodz, Kniaziewicza 1/5, 91-347 Lodz, Poland.
Department of Biophysics of Environmental Pollution, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland.
Int J Mol Sci. 2020 Mar 13;21(6):1953. doi: 10.3390/ijms21061953.
Atherosclerotic plaques are unstable, and their release may result in thrombosis; therefore, currently, antiplatelet therapy with anticoagulants is recommended for the treatment of acute coronary syndrome. The aim of this study was to assess the effect of oxidized cholesterol on human umbilical vascular endothelial cells (HUVECs). The study also examines the protective and repairing effect of dabigatran and rivaroxaban in a model of vascular endothelial damage with 25-hydroxycholesterol (25-OHC).
HUVECs were treated with compounds induce DNA single-strand breaks (SSBs) using the comet assay. Oxidative DNA damage was detected using endonuclease III (Nth) or human 8 oxoguanine DNA glycosylase (hOOG1). Reactive oxygen species (ROS) formation was determined using flow cytometry.
25-hydroxycholesterol caused DNA SSBs, induced oxidative damage and increased ROS in the HUVECs; ROS level was lowered by dabigatran and rivaroxaban. Only dabigatran was able to completely repair the DNA SSBs induced by oxysterol. Dabigatran was able to reduce the level of oxidative damage of pyrimidines induced by oxysterol to the level of control cells.
Observed changes strongly suggest that the tested anticoagulants induced indirect repair of DNA by inhibiting ROS production. Furthermore, dabigatran appears to have a higher antioxidant activity than rivaroxaban.
动脉粥样硬化斑块不稳定，其释放可能导致血栓形成；因此，目前建议对急性冠脉综合征患者采用抗血小板治疗联合抗凝治疗。本研究旨在评估氧化胆固醇对人脐静脉内皮细胞（HUVEC）的影响。本研究还研究了达比加群酯和利伐沙班在 25-羟胆固醇（25-OHC）诱导的血管内皮损伤模型中的保护和修复作用。
采用彗星试验检测化合物诱导 HUVEC 产生 DNA 单链断裂（SSB），用内切酶 III（Nth）或人 8-氧鸟嘌呤 DNA 糖基化酶（hOOG1）检测氧化 DNA 损伤。采用流式细胞术检测活性氧（ROS）的形成。
25-羟胆固醇导致 HUVEC 产生 DNA SSB，诱导氧化损伤并增加 ROS；达比加群酯和利伐沙班降低 ROS 水平。只有达比加群酯能够完全修复由胆甾醇氧化物诱导的 DNA SSB。达比加群酯能够将胆甾醇氧化物诱导的嘧啶氧化损伤水平降低至对照细胞水平。
观察到的变化强烈表明，所测试的抗凝剂通过抑制 ROS 产生诱导了 DNA 的间接修复。此外，达比加群酯的抗氧化活性似乎高于利伐沙班。
