目前，尚无评估脓毒症内皮通透性的临床指标。内皮源性细胞外囊泡（EDEV）正在成为内皮损伤的生物标志物。血小板内皮细胞粘附分子（PECAM）和血管内皮细胞（VE）钙粘蛋白是组成型表达的内皮细胞间粘附分子，调节细胞间粘附和通透性。在此，我们研究了表达细胞间粘附分子（PECAM 或 VE-钙粘蛋白 EDEV）的 EDEV 与内皮通透性和脓毒症严重程度之间的可能关联。 人脐静脉内皮细胞（HUVEC）直接用肿瘤坏死因子 α（TNF-α）刺激或使用血管生成素-1、前列环素或血管内皮生长因子 (VEGF) 等通透性调节剂预处理后，以改变 TNF-α 诱导的内皮通透性过高。使用葡聚糖测定或跨内皮电阻测量内皮渗透性。此外，还进行了一项前瞻性横断面观察研究来分析脓毒症患者的循环 EDEV 水平。使用流式细胞术检查 HUVEC 培养上清液或患者血浆（非败血症，n = 30；败血症，n = 30；败血性休克，n = 42）中的 EDEV。 Wilcoxon 秩和检验用于两组之间的比较。使用 Steel-Dwass 检验进行 3 个或更多组之间的比较。 Spearman 检验用于相关分析。统计显着性设定为 P < .05。TNF-α 刺激 HUVEC 显着增加 EDEV 释放和内皮通透性。血管生成素-1或前列环素预处理可抑制TNF-α诱导的内皮通透性增加，并抑制PECAM和VE-钙粘蛋白EDEV的释放。相反，VEGF预处理增加了TNF-α诱导的内皮通透性以及PECAM和VE-钙粘蛋白EDEV的释放。然而，通透性调节剂预处理并不影响表达炎症刺激诱导的内皮粘附分子（例如E-选择素、细胞内粘附分子-1或血管细胞粘附分子-1）的EDEV的释放。感染性休克组入院时的 PECAM EDEV 数量 (232 [124, 590]/μL) 显着高于脓毒症组 (138 [77,267]/μL) (P = .043)，平均感染性休克组的治疗效果为 98/μL（95% 置信区间 [CI]，2-270/μL），并且 VE-cadherin EDEV 数量（173 [76,339]/μL）也显着更高（P = .004）高于脓毒症组（81 [42,159]/μL），平均治疗效果（ATE）为 79/μL（95% CI，19-171/μL）；这些 EDEV 水平一直保持升高，直到第 5 天。表达细胞间粘附分子（PECAM 或 VE-钙粘蛋白 EDEV）的 EDEV 可能反映内皮通透性增加，并且可能是脓毒症有价值的诊断和预后标志物。版权所有 © 2024 国际麻醉研究协会。

Currently, clinical indicators for evaluating endothelial permeability in sepsis are unavailable. Endothelium-derived extracellular vesicles (EDEVs) are emerging as biomarkers of endothelial injury. Platelet endothelial cell adhesion molecule (PECAM) and vascular endothelial (VE)-cadherin are constitutively expressed endothelial intercellular adhesion molecules that regulate intercellular adhesion and permeability. Herein, we investigated the possible association between EDEVs expressing intercellular adhesion molecules (PECAM+ or VE-cadherin+ EDEVs) and endothelial permeability and sepsis severity.Human umbilical vein endothelial cells (HUVECs) were stimulated with tumor necrosis factor alpha (TNF-α) directly or after pretreatment with permeability-modifying reagents such as angiopoietin-1, prostacyclin, or vascular endothelial growth factor (VEGF) to alter TNF-α-induced endothelial hyperpermeability. Endothelial permeability was measured using the dextran assay or transendothelial electrical resistance. Additionally, a prospective cross-sectional observational study was conducted to analyze circulating EDEV levels in patients with sepsis. EDEVs were examined in HUVEC culture supernatants or patient plasma (nonsepsis, n = 30; sepsis, n = 30; septic shock, n = 42) using flow cytometry. The Wilcoxon rank-sum test was used for comparisons between 2 groups. Comparisons among 3 or more groups were performed using the Steel-Dwass test. Spearman's test was used for correlation analysis. Statistical significance was set at P < .05.TNF-α stimulation of HUVECs significantly increased EDEV release and endothelial permeability. Pretreatment with angiopoietin-1 or prostacyclin suppressed the TNF-α-induced increase in endothelial permeability and inhibited the release of PECAM+ and VE-cadherin+ EDEVs. In contrast, pretreatment with VEGF increased TNF-α-induced endothelial permeability and the release of PECAM+ and VE-cadherin+ EDEVs. However, pretreatment with permeability-modifying reagents did not affect the release of EDEVs expressing inflammatory stimulus-inducible endothelial adhesion molecules such as E-selectin, intracellular adhesion molecule-1, or vascular cell adhesion molecule-1. The number of PECAM+ EDEVs on admission in the septic-shock group (232 [124, 590]/μL) was significantly higher (P = .043) than that in the sepsis group (138 [77,267]/μL), with an average treatment effect of 98/μL (95% confidence interval [CI], 2-270/μL), and the number of VE-cadherin+ EDEVs in the septic-shock group (173 [76,339]/μL) was also significantly higher (P = .004) than that in the sepsis group (81 [42,159]/μL), with an average treatment effect (ATE) of 79/μL (95% CI, 19-171/μL); these EDEV levels remained elevated until day 5.EDEVs expressing intercellular adhesion molecules (PECAM+ or VE-cadherin+ EDEVs) may reflect increased endothelial permeability and could be valuable diagnostic and prognostic markers for sepsis.Copyright © 2024 International Anesthesia Research Society.