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Recombinant human erythropoietin attenuates hepatic injury induced by ischemia/reperfusion in an isolated mouse liver model

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Abstract

Introduction

Apoptosis is a central mechanism of cell death following reperfusion of the ischemic liver. Recombinant human erythropoietin (rhEPO) have an important role in the treatment of myocardial ischemia/reperfusion (I/R) injury, by preventing apoptosis. The aim of the study was to investigate the effect of different regimens of rhEPO in preventing apoptosis following I/R-induced hepatic injury.

Material and methods

Isolated mouse livers were randomly divided into five groups: (1) control group, perfused for the whole study period (105 min); (2) 30-min perfusion followed by 90 min of ischemia and 15 min of reperfusion; (3), (4) and (5) like group 2, but with administration of rhEPO 5,000 units/kg i.p. at 30 min, 24 h, or both 30 min and 24 h respectively, before induction of ischemia. Perfusate liver enzyme levels and intrahepatic caspase-3 activity were measured, and apoptotic cells were identified by morphological criteria, TUNEL assay, and immunohistochemistry for caspase-3. Using immunoblot the expression of the proapoptotic JNK and inhibitor of NFκB (IκBα) were also evaluated. von Willebrand factor (vWF) immunohistochemistry was used as a marker of endothelial cells.

Results

Compared to the I/R livers, all 3 rhEPO pretreated groups showed: a significant reduction in liver enzyme levels (P < 0.05) and intrahepatic caspase-3 activity (P < 0.05), fewer apoptotic hepatocytes (P < 0.05) and positive vWF staining in numerous endothelial cells lining the sinusoids. EPO decreased JNK phosphorylation and the degradation of the inhibitor of NFκB (IκBα) during I/R. There was no added benefit of the multiple- over the single-dose rhEPO regimen.

Conclusion

Pretreatment with one dose of rhEPO can attenuate post-I/R hepatocyte apoptotic liver damage. NFκB and JNK activation is likely to play a pivotal role in the pathophysiology of I/R hepatic injury and might have a key role in EPO-mediated protective effects. This effect is associated with the increase in sinusoidal vWF immunostaining suggests an additional effect of rhEPO in liver angiogenesis recovery. These findings have important implications for the potential use of rhEPO in I/R injury during liver transplantation.

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Authors and Affiliations

  1. Cardiac Research Laboratory of Department of Cardiothoracic Surgery, Rabin Medical Center, Beilinson Campus, Petah Tiqva and Sackler School of Medicine,Tel Aviv University, Tel Aviv, Israel

    Edith Hochhauser, Efrat Kurtzwald & Yelena Cheporko

  2. Department of Histopathology, Rabin Medical Center, Beilinson Campus, Petah Tiqva and Sackler School of Medicine,Tel Aviv University, Tel Aviv, Israel

    Orit Pappo

  3. Department of Surgery B, Rabin Medical Center, Beilinson Campus, Petah Tiqva and Sackler School of Medicine,Tel Aviv University, Tel Aviv, Israel

    Evgeni Ribakovsky & Shlomo Lelchuk

  4. Endocrionology Immunology Laboratory,Felsenstein Medical Research Center, Rabin Medical Center, Beilinson Campus, Petah Tiqva and Sackler School of Medicine,Tel Aviv University, Tel Aviv, Israel

    Amiram Ravid

  5. Liver Institute and Department of Medicine D, Rabin Medical Center, Beilinson Campus, Petah Tiqva and Sackler School of Medicine,Tel Aviv University, Tel Aviv, Israel

    Ziv Ben-Ari

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  1. Edith Hochhauser
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  2. Orit Pappo
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  3. Evgeni Ribakovsky
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  8. Ziv Ben-Ari
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Correspondence to Ziv Ben-Ari.

Additional information

Edith Hochhauser and Orit Pappo are first two coauthors.

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Hochhauser, E., Pappo, O., Ribakovsky, E. et al. Recombinant human erythropoietin attenuates hepatic injury induced by ischemia/reperfusion in an isolated mouse liver model. Apoptosis 13, 77–86 (2008). https://doi.org/10.1007/s10495-007-0155-8

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  • DOI: https://doi.org/10.1007/s10495-007-0155-8

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