title: Alterations in the proteome of pulmonary alveolar type II cells in the rat after hepatic ischemia-reperfusion.
abstract: OBJECTIVE
  Hepatic ischemia-reperfusion can be associated with acute lung injury. Alveolar epithelial type II cells (ATII) play an important role in maintaining lung homeostasis in acute lung injury.
  
  
  METHODS
  To study potentially new mechanisms of hepatic ischemia-reperfusion-induced lung injury, we examined how liver ischemia-reperfusion altered the proteome of ATII.
  
  
  METHODS
  Laboratory investigation.
  
  
  METHODS
  Spontaneously breathing male Zucker rats.
  
  
  METHODS
  Rats were anesthetized with isoflurane. The vascular supply to the left and medial lobe of the liver was clamped for 75 mins and then reperfused. Sham-operated rats were used as controls. After 8 hrs, rats were killed.
  
  
  RESULTS
  Bronchoalveolar lavage and differential cell counts were performed, and tumor necrosis factor-alpha and cytokine-induced neutrophil chemotactic factor-1 in plasma were determined by enzyme-linked immunosorbent assay. ATII were isolated, lysed, tryptically digested, and labeled using isobaric tags (iTRAQ). The samples were fractionated by cation exchange chromatography, separated by high-performance liquid-chromatography, and identified using electrospray tandem mass spectrometry. Spectra were interrogated and quantified using ProteinProspector. Quantitative proteomics provided quantitative data for 94 and 97 proteins in the two groups. Significant changes in ATII protein content included 30% to 40% increases in adenosine triphosphate synthases, adenosine triphosphate/adenosine diphosphate translocase, and catalase (all p < .001). Following liver ischemia-reperfusion, there was also a significant increase in the percentage of neutrophils in bronchoalveolar lavage (48% +/- 26%) compared with sham-operated controls (5% +/- 3%) (p < .01), and plasma tumor necrosis factor-alpha levels were also significantly increased.
  
  
  CONCLUSIONS
  The proteins identified by quantitative proteomics indicated significant changes in moderators of cell metabolism and host defense in ATII. These findings provide new insights into possible mechanisms responsible for hepatic ischemia-reperfusion-related acute lung injury and suggest that ATII cells in the lung sense and respond to hepatic injury.
authors.affiliation: Anesthesia and Perioperative Care, Mass Spectrometry Facility, University of California, San Francisco, CA, USA. hirschj@anesthesia.ucsf.edu
authors.forename: Jan
authors.initials: J
authors.lastname: Hirsch
authors.forename: Claus U
authors.initials: CU
authors.lastname: Niemann
authors.forename: Kirk C
authors.initials: KC
authors.lastname: Hansen
authors.forename: SooJinNa
authors.initials: S
authors.lastname: Choi
authors.forename: Xiao
authors.initials: X
authors.lastname: Su
authors.forename: James A
authors.initials: JA
authors.lastname: Frank
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Fang
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Hirose
authors.forename: Pierre
authors.initials: P
authors.lastname: Theodore
authors.forename: Anil
authors.initials: A
authors.lastname: Sapru
authors.forename: Alma L
authors.initials: AL
authors.lastname: Burlingame
authors.forename: Michael A
authors.initials: MA
authors.lastname: Matthay
mesh_terms: D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D002452:Cell Count; D004734:Energy Metabolism / Q000502:physiology; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D008099:Liver / Q000098:blood supply*; D008297:Male; D040901:Proteomics; D011650:Pulmonary Alveoli / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D011924:Rats, Zucker; D012016:Reference Values; D015427:Reperfusion Injury / Q000235:genetics* / Q000473:pathology