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，造成大量死亡率/发病率，并消耗大量的健康 关怀资源。氢化可的松，抗坏血酸和硫胺素（HAT）治疗已被假设为减少 细胞和器官损伤通过协同作用。一项小型临床HAT试验改善了脓毒症患者的生存率 并引发了对检查机制（S）的兴趣。临床前模型有局限性， 模型还没有产生有效的临床疗法。这些缺陷包括使用内毒素注射， 未能提供常规、有效的脓毒症治疗（抗生素和液体）和极高的死亡率。 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，并且已经预测失败的 干预措施。目前建议的前提来自这样一个假设，即HAT的三个组成部分 协同减少细胞和器官损伤，特别是肾脏。目前的R21提案解决了 通过两个具体目标来定义HAT疗法如何有效的未解决问题。第一个目标将 测量体内氧化应激（OS）的减少和吞噬细胞功能的改善。无创 在脓毒症发作后6小时获得的心率（HR）生理测量准确预测 盲肠结扎穿孔（CLP）模型中的死亡率。远系繁殖的小鼠，无论是雄性还是雌性， 根据HR分层预测存活或死亡，然后随机接受HAT或溶媒， 实验方法既有创新性又有意义。设计是创新的，因为它指导了 治疗对动物可能是有益的，而不是采用典型的一刀切 approach.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中的多个OS参数，以确定HAT是否降低脓毒症患者的OS 小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， 对于脓毒症患者来说，肾脏等重要器官将很难处理。OS将在这些初始 研究作为疗效的标志，但该提案的前提是所有三种HAT成分协同作用， 减少细胞损伤，而不仅仅是减少OS。 脓毒症中的OS和这些细胞将在目标1中用创新技术进行研究。替代治疗， 将测试双重治疗组合，以及不同的剂量和治疗时间后， 败血症由于肾损伤是脓毒症中最常见的功能障碍器官，因此第二个特异性 aim研究了HAT如何减少这种损伤的机制。先进的体内技术将定位 缺氧应激的个体细胞，并且将量化线粒体功能以确定HAT是否改善这一点。 细胞发电站成功完成拟议的研究提供了理解的基础 改善细胞和器官功能的机制。来自R21赠款的数据将用于支持 未来的研究，以确定驱动脓毒症诱导的器官损伤的途径。

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.