TRAUMA-DERIVED EXTRACELLULAR VESICLES ARE SUFFICIENT TO INDUCE ENDOTHELIAL DYSFUNCTION AND COAGULOPATHY.
TRAUMA-DERIVED EXTRACELLULAR VESICLES ARE SUFFICIENT TO INDUCE ENDOTHELIAL DYSFUNCTION AND COAGULOPATHY.
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创伤衍生的细胞外小泡足以诱导内皮功能障碍和凝血病。
DOI：
10.1097/shk.0000000000001950
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发表时间：
2022-07-01
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尽管许多研究已经证明了创伤后细胞外囊泡（EV）的释放增加及其起源差异的变化，但EV的生物学意义尚未得到很好的理解。我们假设创伤/失血性休克（创伤/HS）后释放的EV导致内皮病和凝血病。为了验证这一假设，进行过继转移实验以确定来自休克中严重受伤患者的EV是否足以诱导内皮功能障碍和凝血功能障碍。通过超离心从严重损伤创伤/HS或轻微损伤患者的血浆中富集总EV，并表征其大小和数量。在异氟烷麻醉下，向未受伤的未处理C57 BL/6 J小鼠施用不同浓度的EV，并与接受等体积媒介物（PBS）的小鼠或接受来自最小损伤患者的EV的小鼠进行比较。注射后30分钟，处死小鼠并收集血液用于凝血酶产生（凝血酶-抗凝血酶，达特测定）和通过ELISA测定多配体蛋白聚糖-1。收获肺用于检查组织病理学损伤，并用血管性血友病因子和纤维蛋白共染色以鉴定血管内凝血。从肺中吸出支气管肺泡（BAL）液用于蛋白质测量，作为内皮通透性的指标。数据表示为平均值±SD，p<0.05显著，t检验。在接受不同浓度（5 × 106至3.1 × 109/100 μl/小鼠）的从严重损伤的创伤/HS患者（损伤严重程度评分ISS 34±7）纯化的EV的未处理小鼠中进行初始概念验证实验，并与PBS（对照）小鼠进行比较.在EV输注后30分钟，两组之间的达特和syndecan-1水平均无显著差异。然而，EV组的肺血管通透性和组织病理学损伤显著较高，肺组织显示血管内纤维蛋白沉积。基于该数据，将来自严重创伤/HS患者（ISS 32±6）的EV或来自轻微损伤患者（ISS 8±3）的EV以较高浓度（1 × 109 - 1 × 1010 EV/100 μl/小鼠）给予未处理小鼠。与接受来自最小损伤患者的EV的小鼠相比，创伤/HS EV组的血浆达特和syndecan-1水平显著更高。同样，创伤/HS EV组的BAL蛋白和肺组织病理学损伤更高，肺组织显示血管内纤维蛋白沉积增强。这些数据表明，创伤/HS导致EV的全身释放，其能够诱导内皮病，如通过升高的多配体蛋白聚糖-1和增加的渗透性和凝血病所证明的，如通过增加的达特和血管内纤维蛋白沉积所证明的。针对创伤诱导的电动汽车可能代表了一种新型治疗策略。
Although a number of studies have demonstrated increased release of extracellular vesicles (EVs) and changes in their origin differentials after trauma, the biologic significance of EVs is not well understood. We hypothesized that EVs released after trauma/hemorrhagic shock (trauma/HS) contribute to endotheliopathy and coagulopathy. To test this hypothesis, adoptive transfer experiments were performed to determine whether EVs derived from severely injured patients in shock were sufficient to induce endothelial dysfunction and coagulopathy. Total EVs were enriched from plasma of severely injured trauma/HS or minimally-injured patients by ultracentrifugation and characterized for size and numbers. Under isoflurane anesthesia, non-injured naïve C57BL/6J mice were administered EVs at varying concentrations and compared to mice receiving equal volume vehicle (PBS) or to mice receiving EVs from minimally injured patients. Thirty minutes after injection, mice were sacrificed and blood collected for thrombin generation (thrombin-antithrombin, TAT assay) and syndecan-1 by ELISA. Lungs were harvested for examination of histopathologic injury and co-stained with von Willebrand Factor and fibrin to identify intravascular coagulation. Bronchoalveolar (BAL) fluid was aspirated from lungs for protein measurement as an indicator of the endothelial permeability. Data are presented as mean±SD, p<0.05 significant, t-test. An initial proof- of- concept experiment was performed in naïve mice receiving EVs purified from severely injured trauma/HS patients (injury severity score ISS 34±7) at different concentrations (5x106 to 3.1x109/100 μl/mouse) and compared to PBS (control) mice. Neither TAT nor syndecan-1 levels were significantly different between groups at 30 minutes after EV infusion. However, lung vascular permeability and histopathologic injury were significantly higher in the EV group and lung tissues demonstrated intra-vascular fibrin deposition. Based on this data, EVs from severely injured trauma/HS patients (ISS 32±6) or EVs from minimally injured patients (ISS 8±3) were administered to naïve mice at higher concentrations (1x109 - 1x1010 EV/100 μl/mouse). Compared to mice receiving EVs from minimally injured patients, plasma TAT and syndecan-1 levels were significantly higher in the trauma/HS EV group. Similarly, BAL protein and lung histopathologic injury were higher in the trauma/HS EV group and lung tissues demonstrated enhanced intra-vascular fibrin deposition. These data demonstrate that trauma/HS results in the systemic release of EVs which are capable of inducing endotheliopathy as demonstrated by elevated syndecan-1 and increased permeability and coagulopathy as demonstrated by increased TAT and intra-vascular fibrin deposition. Targeting trauma-induced EVs may represent a novel therapeutic strategy.