title: Amplification of the proinflammatory transcription factor cascade increases with severity of uncontrolled hemorrhage in swine.
abstract: BACKGROUND
  Hypotension causes diffuse liver injury accompanied by increased local production of interleukin-6 (IL-6) in swine models of uncontrolled hemorrhagic shock (HS). IL-6 is transcriptionally up-regulated by nuclear factor (NF)-kappaB and results in activation of signal transducer and activator of transcription-3 (Stat3) in a murine model of controlled HS. Our objectives were: 1). to determine if increased IL-6 production and NF-kappaB and Stat3 activation occurs in a swine model of uncontrolled HS, and 2). to assess whether or not levels of IL-6 mRNA and activity of NF-kappaB and Stat3 correlate with shock severity.
  
  
  METHODS
  Swine were assigned to four groups: 1). control animals (n = 6): no intervention, 2). sham operation (n = 6): celiotomy and splenectomy, 3). uncontrolled hemorrhagic shock (UHS) (n = 6): sham plus grade V vascular liver injury and resuscitation, 4). profound uncontrolled hemorrhagic shock (PUHS) (n = 8): UHS after dilutional hypothermia. Following euthanasia at 2 h, livers were harvested, total RNA isolated, and IL-6 mRNA levels quantified by Q-RT-PCR (ABI Prism 7700, Applied Biosystems International, Foster City, CA). Protein was extracted for measurement of NF-kappaB and Stat3 activity by electrophoretic mobility shift assay (EMSA).
  
  
  RESULTS
  Compared to shams, IL-6 mRNA levels increased 4.5-fold in UHS and 90-fold in PUHS (P < 0.001). Compared with shams; NF-kappaB activity increased 2-fold in both UHS and PUHS (P < 0.05). Stat3 activity was equivalent (not significant) in UHS when compared with shams but increased 5.3-fold in PUHS. (P < 0.05).
  
  
  CONCLUSIONS
  These findings suggest that regional proinflammatory cytokine production results from and perpetuates a proinflammatory transcription factor cascade in a swine model of uncontrolled hemorrhagic shock and indicate that this process is proportional to the severity of shock.
authors.affiliation: Department of Surgery, Stanford University Medical Center, Palo Alto, California 94305, USA. sbrundage@stanford.edu
authors.forename: Susan I
authors.initials: SI
authors.lastname: Brundage
authors.forename: Martin A
authors.initials: MA
authors.lastname: Schreiber
authors.forename: John B
authors.initials: JB
authors.lastname: Holcomb
authors.forename: Nathan
authors.initials: N
authors.lastname: Zautke
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Mastrangelo
authors.forename: Xu
authors.initials: X
authors.lastname: Xq
authors.forename: Joe
authors.initials: J
authors.lastname: Macaitis
authors.forename: David J
authors.initials: DJ
authors.lastname: Tweardy
mesh_terms: D000818:Animals; D004268:DNA-Binding Proteins / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D050796:STAT3 Transcription Factor; D012720:Severity of Illness Index; D012771:Shock, Hemorrhagic / Q000276:immunology*; D013552:Swine; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000276:immunology*
chemical_list: D004268:DNA-Binding Proteins; D015850:Interleukin-6; D016328:NF-kappa B; D050796:STAT3 Transcription Factor; D015534:Trans-Activators; D014157:Transcription Factors