From the Department of Surgery (M.E.D.), University of Wisconsin-Madison School of Medicine, Madison, Wisconsin; Michael and Marian Ilitch Department of Surgery (L.N.D., D.M.L.), Wayne State University School of Medicine, Detroit, Michigan.
J Trauma Acute Care Surg. 2019 Nov;87(5):1061-1069. doi: 10.1097/TA.0000000000002446.
Plasma is an important component of resuscitation after trauma and hemorrhagic shock (T/HS). The specific plasma proteins and the impact of storage conditions are uncertain. Utilizing a microfluidic device system, we studied the effect of various types of plasma on the endothelial barrier function following T/HS.
Human umbilical vein endothelial cells (HUVEC) were cultured in microfluidic plates. The microfluidic plates were subjected to control or shock conditions (hypoxia/reoxygenation + epinephrine, 10 μM). Fresh plasma, 1 day thawed plasma, 5-day thawed plasma and lyophilized plasma were then added. Supplementation of sphingosine-1 phosphate (S-1P) was done in a subset of experiments. Effect on the endothelial glycocalyx was indexed by shedding of syndecan-1 and hyaluronic acid. Endothelial injury/activation was indexed by soluble thrombomodulin, tissue plasminogen activator, plasminogen activator inhibitor-1. Vascular permeability determined by the ratio of angiopoietin-2 to angiopoietin-1. Concentration of S-1P and adiponectin in the different plasma groups was measured.
Human umbilical vein endothelial cells exposed to shock conditions increased shedding of syndecan-1 and hyaluronic acid. Administration of the various types of plasma decreased shedding, except for 5-day thawed plasma. Shocked HUVEC cells demonstrated a profibrinolytic phenotype, this normalized with all plasma types except for 5-day thawed plasma. The concentration of S-1P was significantly less in the 5-day thawed plasma compared with the other plasma types. Addition of S-1P to 5-day thawed plasma returned the benefits lost with storage.
A biomimetic model of the microcirculation following T/HS demonstrated endothelial glycocalyx and endothelial cellular injury/activation as well as a profibrinolytic phenotype. These effects were abrogated by all plasma products except the 5-day thawed plasma. Plasma thawed longer than 5 days had diminished S1-P concentrations. Our data suggest that S1-P protein is critical to the protective effect of plasma products on the endothelial-glycocalyx barrier following T/HS.
血浆是创伤和失血性休克（T/HS）后复苏的重要组成部分。具体的血浆蛋白和储存条件的影响尚不确定。利用微流控装置系统，我们研究了各种类型的血浆对 T/HS 后内皮屏障功能的影响。
将人脐静脉内皮细胞（HUVEC）培养在微流控板上。微流控板接受对照或休克条件（缺氧/复氧+肾上腺素，10 μM）。然后加入新鲜血浆、1 天解冻的血浆、5 天解冻的血浆和冻干血浆。在部分实验中添加了鞘氨醇-1-磷酸（S-1P）。通过硫酸乙酰肝素和透明质酸的脱落来评估内皮糖萼的影响。通过可溶性血栓调节蛋白、组织型纤溶酶原激活物、纤溶酶原激活物抑制剂-1 来评估内皮损伤/激活。血管通透性通过血管生成素-2 与血管生成素-1 的比值来确定。测量不同血浆组中 S-1P 和脂联素的浓度。
暴露于休克条件的人脐静脉内皮细胞增加了硫酸乙酰肝素和透明质酸的脱落。除了 5 天解冻的血浆外，各种类型的血浆均能减少脱落。受冲击的 HUVEC 细胞表现出纤维蛋白溶解的表型，除了 5 天解冻的血浆外，所有血浆类型均使其正常化。与其他血浆类型相比，5 天解冻的血浆中 S-1P 的浓度显著降低。向 5 天解冻的血浆中添加 S-1P 可恢复因储存而丧失的益处。
T/HS 后微循环的仿生模型显示出内皮糖萼和内皮细胞损伤/激活以及纤维蛋白溶解的表型。除了 5 天解冻的血浆外，所有血浆产品都消除了这些作用。解冻时间超过 5 天的血浆中 S1-P 浓度降低。我们的数据表明，S1-P 蛋白对于 T/HS 后血浆产品对内皮糖萼屏障的保护作用至关重要。
