RESUMO
Evidence suggests that light and circadian rhythms profoundly influence the physiologic capacity with which an organism responds to stress. However, the ramifications of light spectrum on the course of critical illness remain to be determined. Here, we show that acute exposure to bright blue spectrum light reduces organ injury by comparison with bright red spectrum or ambient white fluorescent light in two murine models of sterile insult: warm liver ischemia/reperfusion (I/R) and unilateral renal I/R. Exposure to bright blue light before I/R reduced hepatocellular injury and necrosis and reduced acute kidney injury and necrosis. In both models, blue light reduced neutrophil influx, as evidenced by reduced myeloperoxidase (MPO) within each organ, and reduced the release of high-mobility group box 1 (HMGB1), a neutrophil chemotactant and key mediator in the pathogenesis of I/R injury. The protective mechanism appeared to involve an optic pathway and was mediated, in part, by a sympathetic (ß3 adrenergic) pathway that functioned independent of significant alterations in melatonin or corticosterone concentrations to regulate neutrophil recruitment. These data suggest that modifying the spectrum of light may offer therapeutic utility in sterile forms of cellular injury.
Assuntos
Cromoterapia/métodos , Cor , Corticosterona/sangue , Traumatismo por Reperfusão/prevenção & controle , Traumatismo por Reperfusão/fisiopatologia , Animais , Relação Dose-Resposta à Radiação , Proteína HMGB1/sangue , Testes de Função Renal , Testes de Função Hepática , Masculino , Melatonina/sangue , Camundongos , Camundongos Endogâmicos C57BL , Peroxidase/sangue , Doses de Radiação , Traumatismo por Reperfusão/diagnóstico , Índice de Gravidade de Doença , Resultado do TratamentoRESUMO
BACKGROUND: Mortality in sepsis is most often attributed to the development of multiple organ failure. In sepsis, inflammation-mediated endothelial activation, defined as a proinflammatory and procoagulant state of the endothelial cells, has been associated with severity of disease. Thus, the objective of this study was to test the hypothesis that adenosine monophosphate-activated protein kinase (AMPK) activation limits inflammation and endothelium activation to protect against organ injury in sepsis. 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), which is an adenosine monophosphate analog, has been used to upregulate activity of AMPK. Compound C is a cell-permeable pyrrazolopyrimidine compound that inhibits AMPK activity. METHODS: Wild-type mice underwent cecal ligation and puncture (CLP) or sham surgery. Mice were randomized to vehicle, AICAR, or compound C. Mouse kidney endothelial cells were used for in vitro experiments. Renal and liver function were determined by serum cystatin C, blood urea nitrogen (BUN), creatinine, and alanine aminotransferase. Serum cytokines were measured by enzyme-linked immunosorbent assay. Microvascular injury was determined using Evans blue dye and electron microscopy. Immunohistochemistry was used to measure protein levels of phospho-AMPK (p-AMPK), microtubule-associated protein 1A/1B-light chain 3 (LC3), and intracellular adhesion molecule. LC3 levels were used as a measure of autophagosome formation. RESULTS: AICAR decreased liver and kidney injury induced by CLP and minimized cytokine elevation in vivo and in vitro. CLP increased renal and hepatic phosphorylation of AMPK and autophagic signaling as determined by LC3. Inhibition of AMPK with compound C prevented CLP-induced autophagy and exacerbated tissue injury. Additionally, CLP led to endothelial injury as determined by electron microscopy and Evans blue dye extravasation, and AICAR limited this injury. Furthermore, AICAR limited CLP and lipopolysaccharide (LPS)-induced upregulation of intracellular adhesion molecule in vivo and in vitro and decreased LPS-induced neutrophil adhesion in vitro. CONCLUSIONS: In this model, activation of AMPK was protective, and AICAR minimized organ injury by decreasing inflammatory cytokines and endothelial activation. These data suggest that AMPK signaling influences sepsis or LPS-induced endothelial activation and organ injury.
Assuntos
Proteínas Quinases Ativadas por AMP/fisiologia , Inflamação/prevenção & controle , Insuficiência de Múltiplos Órgãos/prevenção & controle , Sepse/complicações , Proteínas Quinases Ativadas por AMP/antagonistas & inibidores , Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/farmacologia , Animais , Autofagia/fisiologia , Adesão Celular , Células Cultivadas , Citocinas/fisiologia , Células Endoteliais/fisiologia , Leucócitos/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ribonucleotídeos/farmacologiaRESUMO
BACKGROUND: Increasing evidence suggests that high fresh frozen plasma:packed red blood cell (FFP:PRBC) and platelet:PRBC (PLT:PRBC) transfusion ratios may prevent or reduce the morbidity associated with early coagulopathy which complicates massive transfusion (MT). We sought to characterize changes in resuscitation which have occurred over time in a cohort severely injured patients requiring MT. METHODS: Data were obtained from a multicenter prospective cohort study evaluating outcomes in blunt injured adults with hemorrhagic shock. MT was defined as requiring ≥10 units PRBCs within 24 hours postinjury. Mean PRBC, FFP, and PLT requirements (per unit; 6 hours, 12 hours, and 24 hours) were determined over time (2004-2009). Sub-MT, those patients just below the threshold for MT, were defined as requiring ≥7 and <10 units PRBCs in the initial 24 hours. The percent of resuscitation given at 6 hours relative to 24 hours total (6 of 24%) was determined and compared across "early" (admission until December 2007) and "recent" (after December 2007) periods for each component. RESULTS: Over the study time period (2004-2009) for the MT group (n = 526), initial base deficit and presenting international normalized ratio were unchanged, while Injury Severity Score was significantly higher. The percent of patients who required MT overall significantly decreased over time. No significant differences were found over time for six-hour, 12-hour, or 24-hour FFP:PRBC and PLT:PRBC transfusion ratios in MT patients. Sub-MT patients (n = 344) had significantly higher six-hour FFP:PRBC ratios and significantly higher six-hour,12-hour, and 24-hour PLT:PRBC ratios in the recent time period. The six h/24 h% total for FFP and PLT transfusion was significantly greater in the recent time period. (FFP: 54% vs.70%; p = 0.004 and PLT 46% vs. 61%; p = 0.048). CONCLUSION: In a severely injured cohort requiring MT, FFP:PRBC and PLT:PRBC ratios have not changed over time, whereas the rate of MT overall has significantly decreased. During the recent time period (after December 2007), significantly higher transfusion ratios and a greater percent of 6-hour/24-hour FFP and PLT were found in the sub-MT group, those patients just below the PRBC transfusion threshold definition of MT. These data suggest early, more aggressive attainment of high transfusions ratios may reduce the requirement for MT and may shift overall blood requirements below those which currently define MT. Further prospective evidence is required to verify these findings.