1 H-NMR Metabolomic Biomarkers of Poor Outcome after

Background: Hemorrhagic shock (HS) following trauma is a leading cause of death among persons under the age of 40. During HS the body undergoes systemic warm ischemia followed by reperfusion during medical intervention. Ischemia/ reperfusion (I/R) results in a disruption of cellular metabolic processes that ultimately lead to tissue and organ dysfunction or failure. Resistance to I/R injury is a characteristic of hibernating mammals. The present study sought to identify circulating metabolites in the rat as biomarkers for metabolic alterations associated with poor outcome after HS. Arctic ground squirrels (AGS), a hibernating species that resists I/R injury independent of decreased body temperature (warm I/R), was used as a negative control. Methodology/principal findings: Male Sprague-Dawley rats and AGS were subject to HS by withdrawing blood to a mean arterial pressure (MAP) of 35 mmHg and maintaining the low MAP for 20 min before reperfusing with Ringers. The animals’ temperature was maintained at 3760.5uC for the duration of the experiment. Plasma samples were taken immediately before hemorrhage and three hours after reperfusion. Hydrophilic and lipid metabolites from plasma were then analyzed via 1 H–NMR from unprocessed plasma and lipid extracts, respectively. Rats, susceptible to I/R injury, had a qualitative shift in their hydrophilic metabolic fingerprint including differential activation of glucose and anaerobic metabolism and had alterations in several metabolites during I/R indicative of metabolic adjustments and organ damage. In contrast, I/R injury resistant AGS, regardless of season or body temperature, maintained a stable metabolic homeostasis revealed by a qualitative 1 H–NMR metabolic profile with few changes in quantified metabolites during HS-induced global I/R. Conclusions/significance: An increase in circulating metabolites indicative of anaerobic metabolism and activation of glycolytic pathways is associated with poor prognosis after HS in rats. These same biomarkers are absent in AGS after HS with warm I/R.

[1]  K. Drew,et al.  Resistance to Systemic Inflammation and Multi Organ Damage after Global Ischemia/Reperfusion in the Arctic Ground Squirrel , 2014, PloS one.

[2]  C. Vandevoort,et al.  Follicle growth, ovulation, and luteal formation in primates and rodents: A comparative perspective , 2013, Experimental biology and medicine.

[3]  C. Ince,et al.  The acute effects of acetate-balanced colloid and crystalloid resuscitation on renal oxygenation in a rat model of hemorrhagic shock. , 2012, Resuscitation.

[4]  L. Hunter,et al.  Kidney proteome changes provide evidence for a dynamic metabolism and regional redistribution of plasma proteins during torpor-arousal cycles of hibernation. , 2012, Physiological genomics.

[5]  David S. Wishart,et al.  MetaboAnalyst 2.0—a comprehensive server for metabolomic data analysis , 2012, Nucleic Acids Res..

[6]  K. Drew,et al.  Neuroprotection: lessons from hibernators. , 2012, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[7]  N. Serkova,et al.  Quantitative analysis in magnetic resonance spectroscopy: from metabolic profiling to in vivo biomarkers. , 2012, Bioanalysis.

[8]  G. Beilman,et al.  Treatment with beta-hydroxybutyrate and melatonin is associated with improved survival in a porcine model of hemorrhagic shock. , 2012, Resuscitation.

[9]  K. Raedschelders,et al.  The cellular and molecular origin of reactive oxygen species generation during myocardial ischemia and reperfusion. , 2012, Pharmacology & therapeutics.

[10]  C. Edelstein,et al.  Renal Protection From Prolonged Cold Ischemia and Warm Reperfusion in Hibernating Squirrels , 2011, Transplantation.

[11]  A. Cariou,et al.  Postcardiac arrest syndrome: from immediate resuscitation to long-term outcome , 2011, Annals of intensive care.

[12]  L. Hunter,et al.  Skeletal muscle proteomics: carbohydrate metabolism oscillates with seasonal and torpor-arousal physiology of hibernation. , 2011, American journal of physiology. Regulatory, integrative and comparative physiology.

[13]  Richard G Melvin,et al.  Deep Sequencing the Transcriptome Reveals Seasonal Adaptive Mechanisms in a Hibernating Mammal , 2011, PloS one.

[14]  Kathleen A Stringer,et al.  The emerging field of quantitative blood metabolomics for biomarker discovery in critical illnesses. , 2011, American journal of respiratory and critical care medicine.

[15]  L. Hunter,et al.  Metabolic cycles in a circannual hibernator. , 2011, Physiological genomics.

[16]  C Loren Buck,et al.  Hibernating above the permafrost: effects of ambient temperature and season on expression of metabolic genes in liver and brown adipose tissue of arctic ground squirrels , 2011, Journal of Experimental Biology.

[17]  C. Nelson,et al.  Global analysis of circulating metabolites in hibernating ground squirrels. , 2010, Comparative biochemistry and physiology. Part D, Genomics & proteomics.

[18]  L. Drewes,et al.  Small-Volume d-&bgr;-Hydroxybutyrate Solution Infusion Increases Survivability of Lethal Hemorrhagic Shock in Rats , 2010, Shock.

[19]  N. Witowski,et al.  Liver metabolomic changes identify biochemical pathways in hemorrhagic shock. , 2010, The Journal of surgical research.

[20]  M. Cohen,et al.  1H-NMR-based metabolic signatures of clinical outcomes in trauma patients--beyond lactate and base deficit. , 2010, The Journal of trauma.

[21]  Sandra L. Martin,et al.  Seasonal protein changes support rapid energy production in hibernator brainstem , 2010, Journal of Comparative Physiology B.

[22]  Hannah V Carey,et al.  Seasonal proteomic changes reveal molecular adaptations to preserve and replenish liver proteins during ground squirrel hibernation. , 2010, American journal of physiology. Regulatory, integrative and comparative physiology.

[23]  I. Netuka,et al.  Gender Differences in Cardiac Ischemic Injury and Protection—Experimental Aspects , 2009, Experimental biology and medicine.

[24]  I. Saul,et al.  Protein kinase C epsilon activation delays neuronal depolarization during cardiac arrest in the euthermic arctic ground squirrel , 2009, Journal of neurochemistry.

[25]  K. Storey Out Cold: Biochemical Regulation of Mammalian Hibernation – A Mini-Review , 2009, Gerontology.

[26]  David S. Wishart,et al.  MetaboAnalyst: a web server for metabolomic data analysis and interpretation , 2009, Nucleic Acids Res..

[27]  Jong M. Rho,et al.  The neuroprotective properties of calorie restriction, the ketogenic diet, and ketone bodies , 2009, Brain Research Reviews.

[28]  Pierre-Gilles Henry,et al.  Adaptive mechanisms regulate preferred utilization of ketones in the heart and brain of a hibernating mammal during arousal from torpor. , 2009, American journal of physiology. Regulatory, integrative and comparative physiology.

[29]  Toru Takahashi,et al.  PREVENTION OF HEMORRHAGIC SHOCK-INDUCED INTESTINAL TISSUE INJURY BY GLUTAMINE VIA HEME OXYGENASE-1 INDUCTION , 2009, Shock.

[30]  S. Kim,et al.  Preliminary study on physiological changes of hemorrhagic shock in rats , 2008, 2008 30th Annual International Conference of the IEEE Engineering in Medicine and Biology Society.

[31]  I. Chaudry,et al.  Bench-to-bedside review: Latest results in hemorrhagic shock , 2008, Critical care.

[32]  Sandra L. Martin,et al.  Proteomic analysis of the winter-protected phenotype of hibernating ground squirrel intestine. , 2008, American journal of physiology. Regulatory, integrative and comparative physiology.

[33]  D. Truong,et al.  Ketogenic diet prevents cardiac arrest-induced cerebral ischemic neurodegeneration , 2008, Journal of Neural Transmission.

[34]  Huiwen W. Zhao,et al.  Arctic Ground Squirrel (Spermophilus Parryii) Hippocampal Neurons Tolerate Prolonged Oxygen—Glucose Deprivation and Maintain Baseline ERK1/2 and JNK Activation Despite Drastic ATP Loss , 2008, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[35]  N. Serkova,et al.  Quantitative analysis of liver metabolites in three stages of the circannual hibernation cycle in 13-lined ground squirrels by NMR. , 2007, Physiological genomics.

[36]  H. Dashti,et al.  Low carbohydrate ketogenic diet enhances cardiac tolerance to global ischaemia , 2007, Acta cardiologica.

[37]  M. T. Andrews,et al.  Advances in molecular biology of hibernation in mammals , 2007, BioEssays : news and reviews in molecular, cellular and developmental biology.

[38]  D. Morrison,et al.  Administration of glutamine after hemorrhagic shock restores cellular energy, reduces cell apoptosis and damage, and increases survival. , 2007, JPEN. Journal of parenteral and enteral nutrition.

[39]  M. Mangino,et al.  Hibernation confers resistance to intestinal ischemia-reperfusion injury. , 2006, American journal of physiology. Gastrointestinal and liver physiology.

[40]  R. Hirose,et al.  Mild hypothermia protects obese rats from fulminant hepatic necrosis induced by ischemia-reperfusion. , 2006, Surgery.

[41]  Ryutaro Hirose,et al.  Metabolic profiling of livers and blood from obese Zucker rats. , 2006, Journal of hepatology.

[42]  K. Drew,et al.  The Arctic Ground Squirrel Brain Is Resistant to Injury From Cardiac Arrest During Euthermia , 2006, Stroke.

[43]  Hannah V. Carey,et al.  Hibernating mammals have enhanced survival and reduced gut damage after hemorrhage , 2006 .

[44]  R. Hirose,et al.  Ischemic Preconditioning Improves Energy State and Transplantation Survival in Obese Zucker Rat Livers , 2005, Anesthesia and analgesia.

[45]  J. Dark,et al.  Annual lipid cycles in hibernators: integration of physiology and behavior. , 2005, Annual review of nutrition.

[46]  Elaine Holmes,et al.  Metabonomic deconvolution of embedded toxicity: application to thioacetamide hepato- and nephrotoxicity. , 2005, Chemical research in toxicology.

[47]  M. Mangino,et al.  Natural resistance to liver cold ischemia-reperfusion injury associated with the hibernation phenotype. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

[48]  Jost Klawitter,et al.  H-NMR-based metabolic signatures of mild and severe ischemia/reperfusion injury in rat kidney transplants. , 2005, Kidney international.

[49]  F. Geiser,et al.  Metabolic rate and body temperature reduction during hibernation and daily torpor. , 2004, Annual review of physiology.

[50]  Sandra L Martin,et al.  Mammalian hibernation: cellular and molecular responses to depressed metabolism and low temperature. , 2003, Physiological reviews.

[51]  P. Rhee,et al.  Ketone and pyruvate Ringer's solutions decrease pulmonary apoptosis in a rat model of severe hemorrhagic shock and resuscitation. , 2003, Surgery.

[52]  Raul Coimbra,et al.  Diagnosis and monitoring of hemorrhagic shock during the initial resuscitation of multiple trauma patients: a review. , 2003, The Journal of emergency medicine.

[53]  C. Buck,et al.  Androgen in free-living arctic ground squirrels: seasonal changes and influence of staged male-male aggressive encounters , 2003, Hormones and Behavior.

[54]  Takashi Sato,et al.  β-hydroxybutyrate, a cerebral function improving agent, protects rat brain against ischemic damage caused by permanent and transient focal cerebral ischemia , 2002 .

[55]  J. McNelis,et al.  Prolonged lactate clearance is associated with increased mortality in the surgical intensive care unit. , 2001, American journal of surgery.

[56]  Kazunori Sato,et al.  Effect of β-Hydroxybutyrate, a Cerebral Function Improving Agent, on Cerebral Hypoxia, Anoxia and Ischemia in Mice and Rats , 2001 .

[57]  K. Frerichs,et al.  Hibernation in Ground Squirrels Induces State and Species-Specific Tolerance to Hypoxia and Aglycemia: An In Vitro Study in Hippocampal Slices , 1998, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[58]  M. Spraul,et al.  750 MHz 1H and 1H-13C NMR spectroscopy of human blood plasma. , 1995, Analytical chemistry.

[59]  L. Sokoloff,et al.  Local Cerebral Blood Flow during Hibernation, a Model of Natural Tolerance to “Cerebral Ischemia” , 1994, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[60]  J. Vion-Dury,et al.  Analysis of plasma lipids by NMR spectroscopy: application to modifications induced by malignant tumors. , 1993, Journal of lipid research.

[61]  B. Barnes Freeze avoidance in a mammal: body temperatures below 0 degree C in an Arctic hibernator. , 1989, Science.

[62]  I. Zucker,et al.  Plasma androgen and gonadotropin levels during hibernation and testicular maturation in golden-mantled ground squirrels. , 1988, Biology of reproduction.

[63]  C. Long,et al.  BIOCHEMICAL STUDIES ON SHOCK : I. THE METABOLISM OF AMINO ACIDS AND CARBOHYDRATE DURING HEMORRHAGIC SHOCK IN THE RAT , 1943 .

[64]  S. Lendemans,et al.  Superiority of acetate compared with lactate in a rodent model of severe hemorrhagic shock. , 2014, The Journal of surgical research.

[65]  Robert Paine,et al.  Metabolic consequences of sepsis-induced acute lung injury revealed by plasma ¹H-nuclear magnetic resonance quantitative metabolomics and computational analysis. , 2011, American journal of physiology. Lung cellular and molecular physiology.

[66]  Daniel S Waterman,et al.  The influence of EDTA and citrate anticoagulant addition to human plasma on information recovery from NMR-based metabolic profiling studies. , 2010, Molecular bioSystems.

[67]  M. Fujimura,et al.  Sputum eosinophilia, airway hyperresponsiveness and airway narrowing in young adults with former asthma. , 2008, Allergology international : official journal of the Japanese Society of Allergology.

[68]  R. Hirose,et al.  Ischemia-reperfusion injury is more severe in older versus young rat livers. , 2007, The Journal of surgical research.

[69]  F. Geiser,et al.  Reduction of metabolism during hibernation and daily torpor in mammals and birds: temperature effect or physiological inhibition? , 2004, Journal of Comparative Physiology B.