title: Effects of rewarming on nuclear factor-kappaB and interleukin 8 expression in cold-preserved alveolar epithelial cells.
abstract: BACKGROUND
  Nuclear factor-kappaB (NF-kappaB) and interleukin (IL)-8 play important roles in the pathophysiology of acute lung injury after lung transplantation. Because alveolar epithelium is one of the most important sites at which IL-8 production takes place after reperfusion of donor lungs, we examined the effects of cold/rewarming on NF-kappaB and IL-8 expression in alveolar epithelial cells.
  
  
  METHODS
  A549 cells were preserved at 4 degrees C for 5 hr and then rewarmed for up to 20 hr. NF-kappaB was analyzed by electrophoretic mobility shift assay. IL-8 mRNA expression was examined by reverse transcription-polymerase chain reaction. IL-8 concentration in the cell culture medium after rewarming was measured by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  NF-kappaB was increased in the nuclear extracts as early as 30 min after rewarming. There was a marked increase in the IL-8 mRNA expression at 1 and 3 hr after rewarming. IL-8 concentration in the cell culture medium was progressively increased during 20 hr following rewarming. The cell culture medium inhibited apoptosis of neutrophils significantly. The cold/rewarming-induced IL-8 production was reduced to approximately 50% by introducing an antisense oligonucleotide for the p65 subunit of NF-kappaB and by treatment with N-acetyl-leucinyl-leucinyl-norleucinal and pyrrolidine dithiocarbamate. The effect of dexamethasone treatment was dose dependent (reduced to approximately 30% at 10-5 M dexamethasone).
  
  
  CONCLUSIONS
  Our results indicate that rewarming of cold-preserved alveolar epithelial cells itself may be an important initiator of the inflammatory cascades, including NF-kappaB activation and IL-8 release. Inhibition of NF-kappaB would be worth trying to control unnecessary IL-8 production and the inflammatory response in the donor lungs.
authors.affiliation: Department of Thoracic Surgery, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan.
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Inoue
authors.forename: Satoshi
authors.initials: S
authors.lastname: Suzuki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kubo
authors.forename: Itaru
authors.initials: I
authors.lastname: Ishida
authors.forename: Shinsaku
authors.initials: S
authors.lastname: Ueda
authors.forename: Takashi
authors.initials: T
authors.lastname: Kondo
mesh_terms: D017209:Apoptosis / Q000276:immunology; D002460:Cell Line; D003080:Cold Temperature; D015870:Gene Expression / Q000276:immunology; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics*; D016040:Lung Transplantation / Q000276:immunology*; D016328:NF-kappa B / Q000378:metabolism*; D009504:Neutrophils / Q000166:cytology; D011650:Pulmonary Alveoli / Q000166:cytology*; D012333:RNA, Messenger / Q000032:analysis; D015427:Reperfusion Injury / Q000276:immunology; D020545:Respiratory Mucosa / Q000166:cytology; D017694:Rewarming; D014021:Tissue Preservation
chemical_list: D016209:Interleukin-8; D016328:NF-kappa B; D012333:RNA, Messenger