Klinik für Anäesthesiologie und Intensivmedizin, Universitätsklinikum Essen, Universität Duisburg-Essen, Hufelandstraße 55, 45122, Essen, Germany.
Institut für Physiologische Chemie, Universitätsklinikum Essen, Universität Duisburg-Essen, Hufelandstraße 55, 45122, Essen, Germany.
Inflammation. 2019 Oct;42(5):1767-1776. doi: 10.1007/s10753-019-01038-6.
In human sepsis, hemolysis is an independent predictor of mortality, but the mechanisms evoking hemolysis have not been fully elucidated. Therefore, we tested the hypotheses that (1) lipopolysaccharide (LPS)-induced hemolysis is dependent on thrombin generation or platelet aggregation and (2) red cell membranes are weakened by LPS. Anesthetized male Wistar rats were subjected to LPS or vehicle for 240 min. The effects of hemostasis inhibition on LPS-induced hemolysis were investigated by use of the thrombin inhibitor argatroban or the platelet function inhibitor eptifibatide. Free hemoglobin concentration, red cell membrane stiffness and red cell morphological changes were determined by spectrophotometry, atomic force microscopy, and light microscopy. Efficacy of argatroban and eptifibatide was assessed by rotational thrombelastometry and impedance aggregometry, respectively. LPS markedly increased free hemoglobin concentration (20.8 μmol/l ± 3.6 vs. 3.5 ± 0.3, n = 6, p < 0.0001) and schistocytes, reduced red cell membrane stiffness, and induced disseminated intravascular coagulation. Inhibition of thrombin formation with argatroban abolished the increase in free hemoglobin concentration, schistocyte formation, and disseminated intravascular coagulation in LPS-treated animals. Eptifibatide had no inhibitory effect. The LPS evoked decrease of red cell stiffness that was not affected by argatroban or eptifibatide. LPS causes hemolysis, schistocyte formation, and red cell membrane weakening in rats. The thrombin inhibitor argatroban but not the platelet inhibitor eptifibatide abolished hemolysis and schistocyte formation. Thus, LPS-induced hemolysis depends on disseminated intravascular coagulation, possibly enhanced by red cell membrane weakening. Clinical studies are necessary to investigate whether thrombin antagonists can decrease hemolysis and mortality in sepsis.
在人类败血症中，溶血是死亡率的独立预测因子，但引起溶血的机制尚未完全阐明。因此，我们检验了以下两个假设：（1）脂多糖（LPS）诱导的溶血依赖于凝血酶生成或血小板聚集；（2）LPS 会削弱红细胞膜。用 LPS 或载体处理麻醉雄性 Wistar 大鼠 240 分钟。通过使用凝血酶抑制剂 argatroban 或血小板功能抑制剂 eptifibatide 来研究止血抑制对 LPS 诱导的溶血的影响。通过分光光度法、原子力显微镜和相差显微镜测定游离血红蛋白浓度、红细胞膜刚性和红细胞形态变化。分别通过旋转血栓弹性测定法和阻抗聚集测定法评估 argatroban 和 eptifibatide 的疗效。LPS 显著增加游离血红蛋白浓度（20.8μmol/l±3.6 与 3.5±0.3，n=6，p<0.0001）和裂片细胞，降低红细胞膜刚性，并诱导弥散性血管内凝血。用 argatroban 抑制凝血酶形成可消除 LPS 处理动物中游离血红蛋白浓度增加、裂片细胞形成和弥散性血管内凝血。eptifibatide 无抑制作用。LPS 引起的红细胞刚性降低不受 argatroban 或 eptifibatide 的影响。LPS 导致大鼠溶血、裂片细胞形成和红细胞膜减弱。凝血酶抑制剂 argatroban 而非血小板抑制剂 eptifibatide 消除了溶血和裂片细胞形成。因此，LPS 诱导的溶血依赖于弥散性血管内凝血，可能通过红细胞膜减弱而增强。需要进行临床研究以探讨凝血酶拮抗剂是否可以降低败血症中的溶血和死亡率。
