Section of General Surgery, Department of Surgery, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Section of General Surgery, Department of Surgery, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Thromb Res. 2020 Jan;185:160-166. doi: 10.1016/j.thromres.2019.11.028. Epub 2019 Nov 26.
During storage, packed red blood cells undergo a series of physical, metabolic, and chemical changes collectively known as the red blood cell storage lesion. One key component of the red blood cell storage lesion is the accumulation of microparticles, which are submicron vesicles shed from erythrocytes as part of the aging process. Previous studies from our laboratory indicate that transfusion of these microparticles leads to lung injury, but the mechanism underlying this process is unknown. In the present study, we hypothesized that microparticles from aged packed red blood cell units induce pulmonary thrombosis.
Leukoreduced, platelet-depleted, murine packed red blood cells (pRBCS) were prepared then stored for up to 14 days. Microparticles were isolated from stored units via high-speed centrifugation. Mice were transfused with microparticles. The presence of pulmonary microthrombi was determined with light microscopy, Martius Scarlet Blue, and thrombocyte stains. In additional studies microparticles were labelled with CFSE prior to injection. Murine lung endothelial cells were cultured and P-selectin concentrations determined by ELISA. In subsequent studies, P-selectin was inhibited by PSI-697 injection prior to transfusion.
We observed an increase in microthrombi formation in lung vasculature in mice receiving microparticles from stored packed red blood cell units as compared with controls. These microthrombi contained platelets, fibrin, and microparticles. Treatment of cultured lung endothelial cells with microparticles led to increased P-selectin in the media. Treatment of mice with a P-selectin inhibitor prior to microparticle infusion decreased microthrombi formation.
These data suggest that microparticles isolated from aged packed red blood cell units promote the development of pulmonary microthrombi in a murine model of transfusion. This pro-thrombotic event appears to be mediated by P-selectin.
在储存过程中，浓缩红细胞会经历一系列物理、代谢和化学变化，这些变化统称为红细胞储存损伤。红细胞储存损伤的一个关键组成部分是微粒的积累，微粒是红细胞衰老过程中脱落的亚微米囊泡。我们实验室的先前研究表明，输注这些微粒会导致肺损伤，但这一过程的机制尚不清楚。在本研究中，我们假设来自陈旧的浓缩红细胞单位的微粒会引起肺血栓形成。
制备去白细胞、血小板 depleted、鼠浓缩红细胞（pRBCS），然后储存最多 14 天。通过高速离心从储存的单位中分离微粒。用微粒给小鼠输血。用光学显微镜、Martius Scarlet Blue 和血小板染色法确定肺微血栓的存在。在其他研究中，用 CFSE 标记微粒，然后进行注射。培养鼠肺内皮细胞，通过 ELISA 测定 P-选择素浓度。在随后的研究中，在输血前用 PSI-697 注射抑制 P-选择素。
与对照组相比，接受来自陈旧浓缩红细胞单位的微粒的小鼠肺部血管中微血栓形成增加。这些微血栓含有血小板、纤维蛋白和微粒。用微粒处理培养的肺内皮细胞会导致培养基中 P-选择素增加。在微粒输注前用 P-选择素抑制剂处理小鼠可减少微血栓形成。
这些数据表明，从陈旧的浓缩红细胞单位中分离的微粒在输血的鼠模型中促进了肺微血栓的形成。这种促血栓形成事件似乎是由 P-选择素介导的。
