Metabolomic Analysis of Hemorrhagic Shock

描述（由申请人提供）：尽管对复苏策略进行了数十年的研究，但平民或军事创伤引起的失血性休克可能发展为不可逆转的休克，每年夺去数千人的生命。向不可逆性发展的原因尚不完全清楚。这项研究的长期目标是更好地确定失血性休克的发病、进展和结局的机制。在海军研究办公室的资助下，我们开发了一种仪器，用于监测严重失血性休克猪模型的组织磷能量学，使用磷核磁共振（NMR）光谱和组织氧指数，使用近红外（NIR）光谱。这些研究表明，在体内磷核磁共振光谱中观察到的磷单酯区域的增加是与失血性休克相关的死亡率的一致的、早期的和独立的标志。结合这些研究，我们存档了动物在失血性休克90分钟后和复苏后的肝脏、肌肉、尿液和血浆样本。我们使用高分辨率核磁共振波谱分析了有限数量的尿液、血浆和组织提取物样本，并检测了在失血性休克和复苏反应中代谢物谱的潜在重要变化。然而，对这些存档的组织和生物流体样本进行全面调查超出了最初提议的范围。在这个R03小额拨款申请中，我们建议分析我们存档的标本的全部补充，并进行必要的对照实验，以确定这些核磁共振光谱中的关键代谢物。我们预计，本申请中提出的研究将使我们能够建立我们现有的全球生理和体内磷能数据之间的关系，这些数据在失血性休克中变化，并从存档组织标本的二次分析中获得更微妙但全面的代谢谱。

DESCRIPTION (provided by applicant): Despite decades of research on resuscitative strategies, hemorrhagic shock from civilian or military trauma can progress to irreversible shock and claims the lives of thousands annually. The cause of progression to irreversibility is incompletely understood. The long term goal of this research is to better define the mechanisms underlying the onset, progression, and outcome of hemorrhagic shock. With funding from the Office of Naval Research, we have developed instrumentation for monitoring tissue phosphoenergetics using phosphorous nuclear magnetic resonance (NMR) spectroscopy and tissue oxygen indices using near infrared (NIR) spectroscopy in a porcine model of severe hemorrhagic shock. These studies show that an observed increase in the phosphomonoester region of in vivo phosphorous NMR spectra is a consistent, early, and independent marker of mortality associated with hemorrhagic shock. In conjunction with these studies we have archived samples of liver, muscle, urine, and plasma from animals at baseline, after 90 minutes of hemorrhagic shock, and following resuscitation. We have analyzed a limited number urine, plasma, and tissue extract samples using high resolution NMR spectroscopy and have detected potentially important changes in metabolite profiles in response to hemorrhagic shock and resuscitation. However, a full investigation of these archived tissue and biofluid samples is beyond the scope of the original proposal. In this R03 Small Grant application we propose to analyze the full complement of our archived specimens and perform the control experiments necessary for identifying key metabolites in these NMR spectra. We anticipate that the studies proposed in this application will allow us to establish a relationship between our existing global physiologic and in vivo phosphoenergetic data that vary in hemorrhagic shock and more subtle but comprehensive metabolic profiles from the secondary analysis of the archived tissue specimens.