Intrahepatic nuclear factor-kappa B activity and alpha 1-acid glycoprotein transcription do not predict outcome after cecal ligation and puncture in the rat.
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.
Chen
J
J
Department of Anesthesia, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
Raj
N
N
Kim
P
P
Andrejko
K M
KM
Deutschman
C S
CS
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
D007375:Interleukin-1; D016328:NF-kappa B; D009961:Orosomucoid; D014409:Tumor Necrosis Factor-alpha