Project 2: Injury & Resuscitation Induced Inflammatory Activation of Innate Im

在重大创伤/失血性休克（T/HS）后，血液制品复苏一直是恢复氧合和促凝潜能的主要方法。由于损伤控制手术的实施提高了患者的总体生存率，急性创伤凝血病（ACoT）已成为导致死亡的主要原因，因为凝血因子和血小板的消耗以及丝氨酸蛋白酶的激活导致血管内皮（ECs）的促炎改变。综上所述，T/HS和ACoT的生理机制似乎是消耗抗蛋白酶，同时激活血浆蛋白酶，通过刺激ECs和中性粒细胞（pmn）上的蛋白酶活化受体（PARS），使损伤患者的表型转变为促炎。这些临床事件使患者易发生pmn介导的损伤后多器官衰竭（MOF）。不加选择地使用血液制品进行复苏可能会导致这种血腥的恶性循环，输血或血浆的数量是死亡率的主要预测指标。储存的血液制品含有生物活性脂质，在体外和体内激活ECs和主要pmn，是急性肺损伤（ALI）的病因，是MOF的一部分。我们的总体假设是，使用储存的血液成分进行的标准复苏忽略了ACoT，并且通过进一步扰乱患者的蛋白质组和脂质组增加了pivin介导的患者发病率。这一假设将通过完成以下具体目标来验证：目的1：研究损伤后患者血浆中积累的蛋白质或复苏过程中输入的血液成分，如蛋白酶和金属酶、抗蛋白酶、磷脂酶、脂质载体和凝血因子。目的2：研究损伤患者复苏过程中输注的促炎脂质、花生四烯酸（AA）及其代谢物存在于输注的血液成分和损伤患者血浆中。目的3：在ALI体内双事件模型中使用这些蛋白和脂质作为第一事件，如果它们激活HMVECs，或者第二个事件，如果它们引起PMN启动。目的4：通过多重细胞因子/趋化因子/生长因子阵列或商用ELISA检测损伤患者复苏前、期间和之后的细胞因子、趋化因子和生长因子，以确定损伤和复苏对体内这些促炎介质浓度的作用。完成这些目标可能会发现改进的复苏方法和更好的受伤患者输血方法，使输血更安全，并优化需要大量输血的受伤患者的生存。

Resuscitation with blood products has been the mainstay of restoring oxygenation and pro-coagulant potential following major trauma/hemorrhagic shock (T/HS). With better overall survival due to the implementation of damage control surgery, the acute coagulopathy of trauma (ACoT) has emerged as a leading cause of mortality through the consumption of clotting factors and platelets and pro-inflammatory changes in the vascular endothelium (ECs) via activation of serine proteases. In sum, the physiologic mechanisms of T/HS and ACoT appear to deplete anti-proteases, while activating plasma proteases, changing the injured patients' phenotype to pro-inflammatory, mediated through stimulation of protease-activated receptors (PARS) on ECs and neutrophils (PMNs). These clinical events predispose patients to PMN-mediated post-injury multiple organ failure (MOF). Indiscriminate resuscitation with blood products may contribute to this bloody vicious cycle and the numbers of blood or plasma transfusions are leading predictors of mortality. Stored blood products contain bioactive lipids which activate ECs and prime PMNs in vitro and in vivo and are etiologic in acute lung injury (ALI), a part of MOF. Our global hypothesis is that standard resuscitation with stored blood components ignores ACoT and by further perturbing the patient's proteome and lipidome increases PIVIN-mediated patient morbidity. This hypothesis will be tested by completion of the following specific aims: Aim 1: Investigate the proteins that accumulate post-injury in patients: plasma or which are infused during resuscitation in transfused blood components, e.g.: proteases and metalloenzymes, anti-proteases, phospholipases, lipid carriers, and coagulation factors. Aim 2: Investigate the pro-inflammatory lipids, arachidonic acid (AA) and it metabolites infused during the resuscitation of injured patients that are present in the transfused blood components and in the injured patient: plasma. Aim 3: Employ these proteins and lipids in a two-event in vivo model of ALI as either the first event, if they activate HMVECs, or the second event, if they cause PMN priming. Aim 4: Measure the cytokines, chemokines, and growth factors in injured patients before, during, and after resuscitation by multiplex cytokine/chemokine/growth factor arrays or commercial ELISA to determine the role of injury and resuscitation on the concentrations of these pro-inflammatory mediators in vivo. Completion of these aims may discover improved methods of resuscitation and better ways to transfuse injured patients to make transfusions safer and to optimize survival for injured patients requiring massive transfusion.