title: Intrahepatic nuclear factor-kappa B activity and alpha 1-acid glycoprotein transcription do not predict outcome after cecal ligation and puncture in the rat.
abstract: OBJECTIVE
  Sepsis is the leading cause of death in critically ill surgical patients. Septic hepatic dysfunction, an important determinant of outcome, is poorly understood but includes inappropriate transcriptional down-regulation. This may be modulated by proinflammatory cytokines. We hypothesized that intrahepatic changes in tumor necrosis factor (TNF)/interleukin (IL)-1-linked processes, such as the activation of the p50 homodimeric and the p65/p50 heterodimeric isoforms of the transcription factor nuclear factor (NF)-kappaB or transcription of the acute phase reactant alpha1-acid glycoprotein (AGP), would correlate with recovery from sepsis.
  
  
  METHODS
  Prospective experimental comparison of sham operation and nonlethal and lethal sepsis in male Sprague-Dawley rats.
  
  
  METHODS
  Nonlethal sepsis was induced by using single-puncture cecal ligation and puncture (CLP). Lethal sepsis was induced via double-puncture CLP. NF-kappaB DNA binding activity was determined by using electrophoretic mobility shift analysis with differentiation of p50/p50 and p50/p65 isoforms by using appropriate antibodies. AGP transcription was assessed with transcription elongation analysis, intrahepatic IL-1beta, and TNF-alpha abundance by using immunohistochemistry, and serum IL-1beta was assessed by using ELISA.
  
  
  RESULTS
  Overall NF-kappaB activity increased equivalently over time after both single- and double-puncture CLP, with a peak occurring 3 hrs after intervention. In single-puncture CLP, there was an increase in the binding of the p50 homodimer form over time. After double-puncture CLP, no such change was observed. AGP transcription was increased equivalently in both models. Intrahepatic IL-1beta was detected 16 and 24 hrs after single-puncture CLP and 6 hrs after double-puncture CLP. After double-puncture CLP, intrahepatic TNF-alpha was detected at 6, 16, and 24 hrs. Serum IL-1beta was undetectable after both single- and double-puncture CLP.
  
  
  CONCLUSIONS
  Although AGP transcription was similar in mild and fulminant sepsis, double-puncture CLP increased the binding activity of the p50 homodimer relative to binding of the p50/p65 NF-kappaB heterodimer. These results imply that transcriptional activity not linked to acute phase responses is an important determinant of outcome in sepsis.
authors.affiliation: Department of Anesthesia, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.forename: N
authors.initials: N
authors.lastname: Raj
authors.forename: P
authors.initials: P
authors.lastname: Kim
authors.forename: K M
authors.initials: KM
authors.lastname: Andrejko
authors.forename: C S
authors.initials: CS
authors.lastname: Deutschman
mesh_terms: D000818:Animals; D002423:Cause of Death; D002432:Cecum / Q000601:surgery*; D004195:Disease Models, Animal; D015536:Down-Regulation / Q000235:genetics* / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D007150:Immunohistochemistry; D007249:Inflammation; D007375:Interleukin-1 / Q000235:genetics / Q000276:immunology; D008026:Ligation; D008099:Liver / Q000276:immunology*; D008297:Male; D016328:NF-kappa B / Q000235:genetics* / Q000276:immunology*; D009961:Orosomucoid / Q000235:genetics* / Q000276:immunology*; D011379:Prognosis; D011446:Prospective Studies; D011677:Punctures; D051381:Rats; D017207:Rats, Sprague-Dawley; D020127:Recovery of Function; D018805:Sepsis / Q000235:genetics* / Q000276:immunology* / Q000401:mortality; D012720:Severity of Illness Index; D015533:Transcriptional Activation / Q000235:genetics* / Q000276:immunology*; D016896:Treatment Outcome; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology
chemical_list: D007375:Interleukin-1; D016328:NF-kappa B; D009961:Orosomucoid; D014409:Tumor Necrosis Factor-alpha