Mechanisms of improved organ function following sepsis treatment with vitamin c, thiamine and hydrocortisone (triple therapy)

脓毒症仍然是一种毁灭性的疾病，会导致大量死亡和发病率，并消耗大量健康 护理资源。氢化可的松、抗坏血酸和硫胺(HAT)疗法被认为可以减少 通过协同作用造成细胞和器官损伤。一项小型临床HAT试验提高了败血症患者的存活率 并引发了人们对审查机制的兴趣(S)。临床前模型有局限性和缺陷的小鼠 模型还没有产生有效的临床治疗方法。这些缺陷包括使用内毒素注射， 未能提供常规、有效的败血症治疗(抗生素和液体)，死亡率非常高。 然而，适当的脓毒症模型确实对已证实的脓毒症治疗有反应，并预测失败。 干预措施。目前提议的前提是假设HAT的三个组成部分 协同作用，减少细胞和器官损伤，特别是肾脏损伤。当前的R21提案针对的是 通过两个具体目标定义HAT疗法如何有效的悬而未决的问题。第一个目标是 在体内测量氧化应激(OS)的减少和吞噬细胞功能的改善。非侵入性 在脓毒症发作后6小时对心率(HR)的生理测量可以准确地预测 盲肠结扎和穿孔(CLP)模型的死亡率。近亲繁殖的老鼠，无论是雄性还是雌性，都将是 根据HR分层预测生存或死亡，然后随机接受帽子或车辆，以及 实验方法既有创新性，又有重要意义。这一设计具有创新性，因为它引导了 对可能有益的动物进行治疗，而不是采用典型的一刀切 接近。该计划意义重大，因为临床试验可以采用同样的方法对患者进行分层。我们 将测量几个不同器官的多个OS参数，以确定HAT是否能减少败血症患者的OS 老鼠。这样的研究在患者中并不真正可行，因为需要实际的组织样本，并且 像肾脏这样的重要器官在败血症患者身上是很难做的。操作系统将在这些初始版本中进行量化 研究作为疗效的标志，但该提议的前提是所有三个HAT组件协同作用以 减少细胞损伤，而不仅仅是减少OS。吞噬细胞功能被认为是由 败血症中的OS和这些细胞将在AIM 1中用创新技术进行研究。 将测试双重治疗组合，以及在发病后不同剂量和时间的治疗 败血症。由于肾损伤是脓毒症中最常见的功能障碍器官，因此第二个特殊的 AIM研究HAT减轻这种损伤的机制(S)。先进的体内技术将本地化 缺氧应激的单个细胞和线粒体功能将被量化，以确定HAT是否改善了这一点 蜂窝发电站。成功完成拟议的研究为理解 改善细胞和器官功能的机制。此R21拨款中的数据将用于支持 未来的研究将确定导致脓毒症所致器官损伤的途径。

Sepsis remains a devastating disease exacting substantial mortality\morbidity and consuming significant health care resources. Hydrocortisone, Ascorbic acid and Thiamine (HAT) therapy has been hypothesized to reduce cell and organ injury through synergistic actions. A small clinical HAT trial improved survival in septic patients and sparked interest into examining mechanism(S). Pre-clinical models have limitations and flawed murine models have not produced effective clinical therapies. These deficiencies include use of endotoxin injections, failure to provide routine, effective sepsis treatments (antibiotics and fluids) and very high mortality rates. However, appropriate sepsis models do respond to proven sepsis treatments and have predicted failed interventions. The premise for the current proposal derives from the hypothesis that the three HAT components synergize to reduce cell and organ injury, especially in the kidney. The current R21 proposal addresses the unresolved issues of defining how HAT therapy is effective through two specific aims. The first aim will measure in vivo reductions in oxidative stress (OS) and improvements in phagocytic cell function. Non-invasive physiologic measurements of heart rate (HR) obtained six hours after the onset of sepsis accurately predict mortality in the cecal ligation and puncture (CLP) model. Outbred mice, both males and females, will be stratified into predicted to live or die based on HR and then randomized to receive HAT or vehicle, an experimental approach with both innovation and significance. The design is innovative since it directs the therapy towards the animals where it may be beneficial, rather than employing the typical one-size-fits-all approach. The plan is significant, since clinical trials could adopt the same approach of stratifying patients. We will measure multiple OS parameters in several different organs to determine if HAT reduces OS in septic mice. Such studies are not really feasible in patients since actual tissue samples are required, and sampling of vital organs such as the kidney would be difficult to do in a septic patient. OS will be quantified in these initial studies as a marker of efficacy, but the premise for the proposal is that all three HAT components synergize to reduce cell injury and not merely reduce OS. Phagocytic cell function has been postulated to be decreased by OS in sepsis and these cells will be studied with innovative techniques in aim 1. Alternative treatments with double therapy combinations will be tested, as well as different doses and times of treatment after the onset of sepsis. Since renal injury is the most frequently observed dysfunctional organ in sepsis, the second specific aim examines the mechanism(s) of how HAT reduces this injury. Sophisticated in vivo techniques will localize hypoxia-stressed individual cells, and mitochondrial function will quantified to determine if HAT improves this cellular powerhouse. Successful completion of the proposed studies provides a foundation for understanding the mechanisms of improved cell and organ function. The data from this R21 grant will be used to support future studies to identify pathways driving sepsis induced organ injury.