Antibiotics improve survival and alter the inflammatory profile in a murine model of sepsis from Pseudomonas aeruginosa pneumonia.

Department of Surgery, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Shock (Impact Factor: 2.73). 05/2003; 19(5):408-14. DOI: 10.1097/
Source: PubMed

ABSTRACT Differing antibiotic regimens can influence both survival and the inflammatory state in sepsis. We investigated whether the addition and/or type of antimicrobial agent could effect mortality in a murine model of Pseudomonas aeruginosa pneumonia-induced sepsis and if antibiotics altered systemic levels of cytokines. FVB/N mice were subjected to intratracheal injection of pathogenic bacteria and were given gentamicin, imipenem, or 0.9% NaCl 2 h after surgery, which continued every 12 h for a total of six doses. Survival at 7 days (n = 24 in each group) was 100% for mice given gentamicin, 88% for mice given imipenem, and 8% for sham mice treated with 0.9% NaCl (P < 0.0001). Systemic interleukin (IL) 6 levels were assayed 6 h postoperatively on all mice to see if they were predictive of outcome. Plasma IL-6 levels above 3,600 pg/mL were associated with a 100% mortality, levels under 1,200 pg/mL were associated with a 100% survival, and levels between 1,200 and 3,600 pg/mL had no utility in predicting mortality. In a separate experiment, mice were sacrificed at 3, 6, 12 or 24 h after instillation of P. aeruginosa and were assayed for levels of TNF-alpha, IL-6, IL-10, and IL-12. Significant alterations in the proinflammatory cytokines TNF-alpha and IL-6 were present at all time points except 3 h between mice treated with antibiotics and sham controls. In contrast, statistically significant differences in the anti-inflammatory cytokine IL-10 were present between the groups only at 6 h, and levels of IL-12 were similar at all time points. These results indicate that both gentamicin and imipenem increase survival at least 10-fold in a model of pneumonia-induced monomicrobial sepsis, and this is predominantly associated with a down-regulation of proinflammatory cytokines.

  • [Show abstract] [Hide abstract]
    ABSTRACT: Sepsis models are frequently based on induction of peritonitis, with cecal ligation and puncture reflecting the prototypical model. However, there is an ongoing discussion about the limitations of these models due to their variability in progression and outcome. Since standardization is a cornerstone of experimental models, we aimed to develop a reliable and reproducible procedure for induction of peritonitis. A human stool batch was processed for -80° storage. For induction of peritonitis in fluid-resuscitated rats, a defined volume of stool suspension from this batch was injected intraperitoneally. For characterization of the model, physiologic and inflammatory changes were evaluated after sepsis induction. Survival analyses with the same batch were repeated in four independent experiments over a time period of 16 mo. The polymicrobial infection resulted in severe peritoneal inflammation with a systemic increase in cytokines. The mortality rate at 15 h was 29% and this was reproducible over a 16 mo time period. If antibiotic treatment was applied, a 50% survival was achieved. Laboratory markers indicated a progressive multi-organ dysfunction, while blood gas analysis showed respiratory compensation of a metabolic acidosis, and maintenance of PaO(2). Intravital microscopy of the liver revealed an impaired microcirculation. A decreased hemostatic potential was demonstrated by rotational thromboelastometry. Despite clinical recovery within 3 d, surviving animals showed laboratory and histologic signs of persisting inflammation even after 2 wk. This model reflects many features of human sepsis. Application of an infectious focus that is both quantitatively and qualitatively defined assures high reproducibility. Moreover, the procedure is simple and can be easily standardized.
    Journal of Surgical Research 09/2011; 170(1):e123-34. DOI:10.1016/j.jss.2011.05.019 · 2.12 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: To determine the relationship between sublingual and intestinal mucosal microcirculatory perfusion. Observational, experimental study. University-affiliated large animal laboratory. Ten fasted, anesthetized, mechanically ventilated, male pigs randomized to a sham group (n = 3) or to a hyperdynamic septic shock group (n = 7) in which cholangitis was induced by direct infusion of Escherichia coli into the common bile duct. This model was developed because it is not accompanied by changes in intra-abdominal pressure. The sublingual and intestinal microcirculations were simultaneously assessed at 4-hr intervals for up to 12 hrs with a modified orthogonal polarization spectral device and functional microvessel density and erythrocyte velocity were measured quantitatively. In sham animals, both regions maintained a stable functional microvessel density and erythrocyte velocity throughout the study period. In contrast, in septic animals, already after 4 hrs of sepsis, functional microvessel density was markedly decreased (>50%) in the sublingual and gut regions; mean erythrocyte velocity decreased dramatically and similarly in both regions, from 1022 +/- 80 to 265 +/- 43 mum/sec in the sublingual region and from 1068 +/- 45 to 243 +/- 115 mum/sec in the gut (p < 0.001, at T12). There was a significant correlation between the sublingual and gut microcirculations in septic animals (r = 0.92, p < 0.0001). The severity and the time course of microcirculatory changes were similar in the sublingual and in the gut region in this clinically relevant model of severe sepsis. These findings support the sublingual region as an appropriate region to monitor the microcirculation in sepsis.
    Critical care medicine 09/2009; 37(11):2875-81. DOI:10.1097/CCM.0b013e3181b029c1 · 6.15 Impact Factor