title: Identification of differentially expressed genes after partial rat liver ischemia/reperfusion by suppression subtractive hybridization.
abstract: OBJECTIVE
  To identify potential diagnostic target genes in early reperfusion periods following warm liver ischemia before irreversible liver damage occurs.
  
  
  METHODS
  We used two strategies (SSH suppression subtractive hybridization and hybridization of cDNA arrays) to determine early changes in gene expression profiles in a rat model of partial WI/R, comparing postischemic and adjacent nonischemic liver lobes. Differential gene expression was verified (WI/R; 1 h/2 h) and analyzed in more detail after warm ischemia (1 h) in a reperfusion time kinetics (0, 1, 2 and 6 h) and compared to untreated livers by Northern blot hybridizations. Protein expression was examined on Western blots and by immunohistochemistry for four differentially expressed target genes (Hsp70, Hsp27, Gadd45a and IL-1rI).
  
  
  RESULTS
  Thirty-two individual WI/R target genes showing altered RNA levels after confirmation by Northern blot analyzes were identified. Among them, six functionally uncharacteristic expressed sequences and 26 known genes (12 induced in postischemic liver lobes, 14 with higher transcriptional expression in adjacent nonischemic liver lobes). Functional categories of the verified marker genes indicate on the one hand cellular stress and tissue damage but otherwise activation of protective cellular reactions (AP-1 transcription factors, apoptosis related genes, heat shock genes). In order to assign the transcriptional status to the biological relevant protein level we demonstrated that Hsp70, Hsp27, Gadd45a and IL-1rI were clearly up-regulated comparing postischemic and untreated rat livers, suggesting their involvement in the WI/R context.
  
  
  CONCLUSIONS
  This study unveils a WI/R response gene set that will help to explore molecular pathways involved in the tissue damage after WI/R. In addition, these genes especially Hsp70 and Gadd45a might represent promising new candidates indicating WI/R liver damage.
authors.affiliation: Institute of Molecular Pathology, University of Heidelberg, Heidelberg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Fallsehr
authors.forename: Christina
authors.initials: C
authors.lastname: Zapletal
authors.forename: Michael
authors.initials: M
authors.lastname: Kremer
authors.forename: Resit
authors.initials: R
authors.lastname: Demir
authors.forename: Magnus
authors.initials: M
authors.lastname: von Knebel Doeberitz
authors.forename: Ernst
authors.initials: E
authors.lastname: Klar
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods*; D055551:HSP27 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006360:Heat-Shock Proteins / Q000235:genetics; D006358:Hot Temperature; D007700:Kinetics; D008099:Liver / Q000473:pathology / Q000502:physiology*; D009363:Neoplasm Proteins / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D017472:Receptors, Interleukin-1 / Q000235:genetics; D053573:Receptors, Interleukin-1 Type I; D015427:Reperfusion Injury / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology*; D018808:Transcription Factor AP-1 / Q000502:physiology; D014158:Transcription, Genetic / Q000502:physiology; D000097642:GADD45 Proteins
chemical_list: D018797:Cell Cycle Proteins; D055551:HSP27 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D017472:Receptors, Interleukin-1; D053573:Receptors, Interleukin-1 Type I; D018808:Transcription Factor AP-1; C499073:Gadd45a protein, rat; D000097642:GADD45 Proteins; C486114:Hspb1 protein, rat