Hepatic cryoablation-induced multisystem injury: bioluminescent detection of NF-kappaB activation in a transgenic mouse model.
Hepatic injury from cryoablation has been associated with multisystem injury, including adult respiratory distress syndrome, renal insufficiency, and coagulopathy; but the responsible mechanisms have not been well defined. In the present study we investigated the role of the transcription factor NF-kappaB in the multiorgan inflammatory response to hepatic cryoablation utilizing a novel in vivo system for determining NF-kappaB activity. Using transgenic mice expressing photinus luciferase under the control of the 5' HIV-LTR (an NF-kappaB-dependent promoter), we measured luciferase activity in the liver, lungs, and kidneys as a marker for NF-kappaB activity. Luciferase production was determined by in vivo bioluminescence and by luciferase assays of tissue homogenates. After measurement of basal luciferase activity, mice were treated with 35% hepatic cryoablation or sham laparotomy and injected with luciferin (0.75 mg/mouse). Photon emission from the liver, lungs, and kidneys was measured at multiple time points. Hepatic cryoablation induced a significant increase in photon emission by the liver, lungs, and kidneys, which correlated with markedly increased luciferase activity measured from each organ after death. Lung lavage 4 hours after cryoablation showed neutrophilic lung inflammation with increased MIP-2 levels compared with sham surgery. These findings demonstrate that 35% hepatic cryoablation is associated with NF-kappaB activation in the remnant liver and multiple distant sites, and may be causally related to the multisystem injury that is seen after direct liver injury.
Sadikot
Ruxanna T
RT
Department of Medicine, Division of Allergy, Pulmonary, and Critical Care Medicine, Nashville, TN, USA.
Wudel
L James
LJ
Jansen
Duco E
DE
Debelak
Jacob P
JP
Yull
Fiona E
FE
Christman
John W
JW
Blackwell
Timothy S
TS
Chapman
William C
WC
D000818:Animals; D003452:Cryosurgery / Q000009:adverse effects* / Q000379:methods; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D007668:Kidney / Q000378:metabolism / Q000503:physiopathology; D008099:Liver / Q000378:metabolism / Q000503:physiopathology / Q000601:surgery*; D008163:Luminescent Measurements; D008168:Lung / Q000378:metabolism / Q000503:physiopathology; D051379:Mice; D008822:Mice, Transgenic; D009102:Multiple Organ Failure / Q000209:etiology* / Q000503:physiopathology*; D016328:NF-kappa B / Q000032:analysis / Q000378:metabolism*; D018570:Risk Assessment; D012680:Sensitivity and Specificity
D016328:NF-kappa B