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. 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 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. MAIN 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.
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. 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 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. MAIN 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: Roman Pfeifer; Sophie Darwiche; Lauryn Kohut; Timothy R Billiar; Hans-Christoph Pape Journal: Clin Orthop Relat Res Date: 2013-09 Impact factor: 4.176