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Identification of differentially expressed genes after partial rat liver ischemia/reperfusion by suppression subtractive hybridization.

ABSTRACT: AIM: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. CONCLUSION: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.

SUBMITTER: Fallsehr C

PROVIDER: S-EPMC4250677 | biostudies-literature | 2005 Mar

REPOSITORIES: biostudies-literature

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Identification of differentially expressed genes after partial rat liver ischemia/reperfusion by suppression subtractive hybridization.

Fallsehr Christine C Zapletal Christina C Kremer Michael M Demir Resit R von Knebel Doeberitz Magnus M Klar Ernst E

World journal of gastroenterology 20050301 9

<h4>Aim</h4>To identify potential diagnostic target genes in early reperfusion periods following warm liver ischemia before irreversible liver damage occurs.<h4>Methods</h4>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 ...[more]

PMID: 15761968

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Identification of differentially expressed genes after partial rat liver ischemia/reperfusion by suppression subtractive hybridization.

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Identification of differentially expressed genes after partial rat liver ischemia/reperfusion by suppression subtractive hybridization.

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