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

血液产品复苏一直是恢复严重创伤/失血性休克(T/HS)后氧合和促凝血潜能的主要方法。随着损伤控制手术的实施，创伤的急性凝固性病变(ACoT)已经成为导致死亡的主要原因，它通过消耗凝血因子和血小板，以及通过激活丝氨酸蛋白酶导致血管内皮细胞(ECs)的促炎变化。综上所述，T/HS和ACoT的生理机制似乎是通过刺激内皮细胞和中性粒细胞上的蛋白酶激活受体(PARs)，在激活血浆蛋白水解酶的同时，将损伤患者的表型转变为促炎状态。这些临床事件使患者容易发生PMN介导的损伤后多器官功能衰竭(MOF)。不分青红皂白地用血液制品复苏可能会导致这种血腥的恶性循环，而输血或血浆的数量是死亡率的主要预测因素。储存的血液制品含有生物活性脂质，在体外和体内激活内皮细胞和启动PMN，是MOF的一部分，是急性肺损伤(ALI)的病因。我们的全球假设是，标准的储存血液成分复苏忽略了ACoT，并通过进一步干扰患者的蛋白质组和脂肪组会增加Pivin介导的患者发病率。这一假说将通过完成以下特定目标来验证：目的1：调查在患者受伤后积聚的蛋白质：血浆或在复苏过程中在输血成分中输注的蛋白质，例如：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：研究创伤患者复苏过程中输注的促炎性脂质、花生四烯酸(AA)及其代谢物在输血成分和受伤患者血浆中的分布。目的3：在ALI的两事件体内模型中，使用这些蛋白质和脂质作为第一事件(如果它们激活HMVECs)或第二事件(如果它们引起PMN启动)。目的：采用复合细胞因子/趋化因子/生长因子芯片或商业酶联免疫吸附试验检测创伤患者复苏前、复苏中和复苏后细胞因子、趋化因子和生长因子的变化，以确定损伤和复苏对体内这些促炎介质浓度的影响。完成这些目标可能会发现更好的复苏方法和更好的方法来为受伤患者输血，以使输血更安全，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.