Intrahepatic nuclear factor-kappa B activity and alpha(1)-acid glycoprotein transcription do not predict outcome after cecal ligation and puncture inthe 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-regula tion. This may be modulated by proinflammatory cytokines. We hypothesized t hat 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 alpha (1)-acid glycopr otein (AGP), would correlate with recovery from sepsis. Design: prospective experimental comparison of sham operation and nonlethal and lethal sepsis in male Sprague-Dawley rats. Interventions Nonlethal sepsis was induced by using single-puncture cecal l igation and puncture (CLP), Lethal sepsis was induced via double-puncture C LP, NF-kappaB DNA binding activity was determined by using electrophoretic mobility shift analysis with differentiation of p50/p50 and p50/p65 isoform s by using appropriate antibodies. AGP transcription was assessed with tran scription elongation analysis, intrahepatic IL-1 beta, and TNF-alpha abunda nce by using immunohistochemistry, and serum IL-1 beta was assessed by usin g ELISA. Main Results: Overall NF-kappaB activity increased equivalently over time a fter both single- and double-puncture CLP, with a peak occurring 3 hrs afte r intervention. In single-puncture CLP, there was an increase in the bindin g of the p50 homodimer form over time. After double-puncture CLP, no such c hange was observed. AGP transcription was increased equivalently in both mo dels. Intrahepatic IL-1 beta was detected 16 and 24 hrs after single-punctu re CLP and 6 hrs after double-puncture CLP. After double-puncture CLP, intr ahepatic TNF-alpha was detected at 6, 16, and 24 hrs. Serum IL-1 beta was u ndetectable after both single- and double-puncture CLP. Conclusions: Although AGP transcription was similar in mild and fulminant s epsis, double-puncture CLP increased the binding activity of the p50 homodi mer 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.