Plasma obtained during human endotoxemia increases endothelial albumin permeability in vitro
Plasma obtained during human endotoxemia increases endothelial albumin permeability in vitro
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DOI：
10.1097/01.shk.0000209527.35743.b0
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发表时间：
2006-04-01
期刊：
影响因子：
3.1
通讯作者：
Pickkers, P
中科院分区：
文献类型：
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作者：
van Eijk, LT;Nooteboom, A;Pickkers, P
为了深入了解败血症期间血管通透性增加的发病机制，我们研究了人类实验性内毒素血症期间获得的血浆对培养的单层内皮细胞通透性的影响。八名健康受试者接受了静脉注射。剂量为 2 ng/kg 大肠杆菌 O:113 脂多糖 (LPS)。随着时间的推移，测量了可能诱导血管通透性的各种血浆介质的浓度。将LPS给药之前、2小时和4小时后获得的血浆添加到在半透膜上培养的人脐静脉内皮细胞中。测定内皮单层对异硫氰酸荧光素标记的牛血清白蛋白的渗透性，并表示为与跨空 Transwell-COL（Corning Life Sciences B.V.，Schiphol-Rijk，荷兰）膜（即没有内皮单层）测量的相对浓度相比，异硫氰酸荧光素-牛血清白蛋白的相对浓度。渗透性水平与各种介质的浓度相关。实验性内毒素血症导致肿瘤坏死因子 a、白细胞介素 (IL) 1β、IL-6、IL-8、IL-10 和血管内皮生长因子水平升高，以及 IL-12 和 IFN-γ 适度增加（所有 P 值 < 0.01）。给予LPS后2和4小时获得的血浆与人脐静脉内皮细胞一起孵育，使相对渗透性从基线水平（中值）17%（范围，14% - 31%）分别增加到23%（范围，12% - 39%；P=不显着）和28%（范围，11%-40%；P < 0.05）。血管内皮生长因子和 IL-10（而非 TNF-α 或任何其他介质）的血浆水平与内皮通透性的增加显着相关（分别为 r = 0.47，P = 0.038；r = 0.43，P = 0.038）。这里提供的数据表明，从实验性人内毒素血症中获得的血浆可增加体外内皮白蛋白的通透性。因此，培养的人内皮单层提供了研究脓毒症相关血管变化的模型。
To gain insight in the pathogenesis of increased vascular permeability during sepsis, we studied the effect of plasma obtained during human experimental endotoxemia on the permeability of cultured endothelial monolayers. Eight healthy subjects received an i.v. dose of 2 ng/kg Escherichia coli O:113 lipopolysaccharide (LPS). The concentration of various plasma mediators that supposedly induce vascular permeability was measured over time. Plasmas that were obtained before, and 2 and 4 h after the administration of LPS were added to human umbilical venular endothelial cells that were cultured on semipermeable membranes. The permeability of the endothelial monolayers to fluorescein isothiocyanate-labeled bovine serum albumin was determined and expressed as the relative concentration of fluorescein isothiocyanate-bovine serum albumin when compared with that measured across empty Transwell-COL (Corning Life Sciences B.V., Schiphol-Rijk, The Netherlands) membranes (i.e., without endothelial monolayers). The permeability levels were correlated with the concentrations of various mediators. Experimental endotoxemia resulted in elevated levels of tumor necrosis factor a, interleukin (IL) 1 beta, IL-6, IL-8, IL-10, and vascular endothelial growth factor and a moderate increase of IL-12 and IFN-gamma (all Pvalues < 0.01). Incubation of human umbilical venular endothelial cells with plasma obtained 2 and 4 h after the administration of LPS increased the relative permeability from a baseline level (median) of 17% (range, 14% - 31%) to 23% (range, 12% - 39%; P = not significant) and 28% (range, 11%-40%; P < 0.05), respectively. Plasma levels of vascular endothelial growth factor and IL-10, but not TNF-alpha or any other mediators, significantly correlated with the increase in endothelial permeability (r = 0.47, P = 0.038; r = 0.43, P = 0.038, respectively). The data presented here demonstrate that plasmas obtained from experimental human endotoxemia increase endothelial albumin permeability in vitro. Thus, cultured human endothelial monolayers provide a model to study sepsis-associated vascular changes.