Neutrophil Extracellular Traps Promote Thrombin Generation Through Platelet-Dependent and Platelet-Independent Mechanisms
Neutrophil Extracellular Traps Promote Thrombin Generation Through Platelet-Dependent and Platelet-Independent Mechanisms
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DOI：
10.1161/atvbaha.114.304114
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发表时间：
2014-09-01
影响因子：
8.7
通讯作者：
Liaw, Patricia C.
中科院分区：
文献类型：
--
作者：
Gould, Travis J.;Vu, Trang T.;Liaw, Patricia C.
中性粒细胞被微生物或炎症刺激物激活，导致由DNA、组蛋白和抗菌蛋白组成的中性粒细胞胞外陷阱（NET）释放。在纯化的系统中，无细胞DNA（CFDNA）激活凝血的内在途径，而组蛋白通过血小板依赖性机制促进凝血酶生成。然而，CFDNA/组蛋白复合物作为完整NET的一部分的总体促凝血作用是未知的。在这项研究中，我们研究了从活化的中性粒细胞释放的完整NET的促凝血潜力。我们还确定了CFDNA和组蛋白的相对贡献，凝血酶产生血浆从patients with sepsis.Approach and Results-NET释放从佛波酯肉豆蔻酸激活的中性粒细胞增强凝血酶生成血小板贫血浆。这种效应是DNA依赖性的（通过DNA酶处理证实），并通过内源性凝血途径发生（用凝血因子XII和凝血因子XI耗竭血浆证实）。在用玉米胰蛋白酶抑制剂处理的富含血小板的血浆中，添加肉豆蔻酸佛波酯激活的中性粒细胞增加凝血酶的产生并缩短Toll样受体2和Toll样受体4依赖性机制的滞后时间。添加DNA酶进一步增强凝血酶生成，表明拆除NET支架增加组蛋白介导的血小板依赖性凝血酶生成。在血小板贫乏的血浆样本与败血症患者，我们发现内源性CFDNA和凝血酶生成之间的正相关性，并添加DNase衰减thrombin generation. Conclusions这些研究检查CFDNA和组蛋白的促凝血活性的背景下，NET。我们的研究还暗示了凝血的内在途径在脓毒症发病机制中的作用。
Objective-Activation of neutrophils by microbial or inflammatory stimuli results in the release of neutrophil extracellular traps (NETs) that are composed of DNA, histones, and antimicrobial proteins. In purified systems, cell-free DNA (CFDNA) activates the intrinsic pathway of coagulation, whereas histones promote thrombin generation through platelet-dependent mechanisms. However, the overall procoagulant effects of CFDNA/histone complexes as part of intact NETs are unknown. In this study, we examined the procoagulant potential of intact NETs released from activated neutrophils. We also determined the relative contribution of CFDNA and histones to thrombin generation in plasmas from patients with sepsis.Approach and Results-NETs released from phorbyl myristate-activated neutrophils enhance thrombin generation in platelet-poor plasma. This effect was DNA dependent (confirmed by DNase treatment) and occurred via the intrinsic pathway of coagulation (confirmed with coagulation factor XII- and coagulation factor XI-depleted plasma). In platelet-rich plasma treated with corn trypsin inhibitor, addition of phorbyl myristate-activated neutrophils increased thrombin generation and shortened the lag time in a toll-like receptor-2- and toll-like receptor-4-dependent mechanism. Addition of DNase further augmented thrombin generation, suggesting that dismantling of the NET scaffold increases histone-mediated, platelet-dependent thrombin generation. In platelet-poor plasma samples from patients with sepsis, we found a positive correlation between endogenous CFDNA and thrombin generation, and addition of DNase attenuated thrombin generation.Conclusions-These studies examine the procoagulant activities of CFDNA and histones in the context of NETs. Our studies also implicate a role for the intrinsic pathway of coagulation in sepsis pathogenesis.