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

被引:10
作者
Fallsehr, Christine [1 ]
Zapletal, Christina [2 ]
Kremer, Michael [3 ]
Demir, Resit [4 ]
Doeberitz, Magnus von Knebel [1 ]
Klar, Ernst [5 ]
机构
[1] Heidelberg Univ, Inst Mol Pathol, D-69120 Heidelberg, Germany
[2] Goethe Univ Frankfurt, Dept Surg, D-6000 Frankfurt, Germany
[3] Heidelberg Univ, Dept Surg, D-6900 Heidelberg, Germany
[4] Univ Erlangen Nurnberg, Dept Surg, Erlangen, Germany
[5] Univ Rostock, Dept Surg, D-2500 Rostock 1, Germany
关键词
Organ injury; Liver ischemia; AP-1 transcription factor; Heat shock protein; Gadd45A; Suppression subtractive hybridization;
D O I
10.3748/wjg.v11.i9.1303
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
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. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved.
引用
收藏
页码:1303 / 1315
页数:13
相关论文
共 36 条
  • [1] REPERFUSION INJURY TO ENDOTHELIAL-CELLS FOLLOWING COLD ISCHEMIC STORAGE OF RAT LIVERS
    CALDWELLKENKEL, JC
    CURRIN, RT
    TANAKA, Y
    THURMAN, RG
    LEMASTERS, JJ
    [J]. HEPATOLOGY, 1989, 10 (03) : 292 - 299
  • [2] Recent insights on the mechanisms of liver preconditioning
    Carini, R
    Albano, E
    [J]. GASTROENTEROLOGY, 2003, 125 (05) : 1480 - 1491
  • [3] GENOMIC SEQUENCING
    CHURCH, GM
    GILBERT, W
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (07): : 1991 - 1995
  • [4] HEPATIC INTERCELLULAR COMMUNICATION IN SHOCK AND INFLAMMATION
    CLEMENS, MG
    BAUER, M
    GINGALEWSKI, C
    MIESCHER, E
    ZHANG, J
    [J]. SHOCK, 1994, 2 (01): : 1 - 9
  • [5] Lung and liver injury following hepatic ischemia/reperfusion in the rat is increased by exogenous lipopolysaccharide which also increases hepatic TNF production in vivo and in vitro
    Colletti, LM
    Green, M
    [J]. SHOCK, 2001, 16 (04): : 312 - 319
  • [6] Suppression subtractive hybridization: A method for generating differentially regulated or tissue-specific cDNA probes and libraries
    Diatchenko, L
    Lau, YFC
    Campbell, AP
    Chenchik, A
    Moqadam, F
    Huang, B
    Lukyanov, S
    Lukyanov, K
    Gurskaya, N
    Sverdlov, ED
    Siebert, PD
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (12) : 6025 - 6030
  • [7] Doi Y, 2001, HEPATO-GASTROENTEROL, V48, P533
  • [8] Therapeutic approaches for ischemia/reperfusion injury in the liver
    Fan, CG
    Zwacka, RM
    Engelhardt, JF
    [J]. JOURNAL OF MOLECULAR MEDICINE-JMM, 1999, 77 (08): : 577 - 592
  • [9] Flohe S, 1998, TRANSPL INT, V11, P89
  • [10] Mechanism of cell death during warm hepatic ischemia-reperfusion in rats: Apoptosis or necrosis?
    Gujral, JS
    Bucci, TJ
    Farhood, A
    Jaeschke, H
    [J]. HEPATOLOGY, 2001, 33 (02) : 397 - 405