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BMC Gastroenterology
Published in: BMC Gastroenterology 1/2009

Open Access 01-12-2009 | Research article

No evidence for protective erythropoietin alpha signalling in rat hepatocytes

Authors: Thorsten Bramey, Patricia Freitag, Joachim Fandrey, Ursula Rauen, Katja Pamp, Jochen Erhard, Stilla Frede, Herbert de Groot, Frank Petrat

Published in: BMC Gastroenterology | Issue 1/2009

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Abstract

Background

Recombinant human erythropoietin alpha (rHu-EPO) has been reported to protect the liver of rats and mice from ischemia-reperfusion injury. However, direct protective effects of rHu-EPO on hepatocytes and the responsible signalling pathways have not yet been described. The aim of the present work was to study the protective effect of rHu-EPO on warm hypoxia-reoxygenation and cold-induced injury to hepatocytes and the rHu-EPO-dependent signalling involved.

Methods

Loss of viability of isolated rat hepatocytes subjected to hypoxia/reoxygenation or incubated at 4°C followed by rewarming was determined from released lactate dehydrogenase activity in the absence and presence of rHu-EPO (0.2–100 U/ml). Apoptotic nuclear morphology was assessed by fluorescence microscopy using the nuclear fluorophores H33342 and propidium iodide. Erythropoietin receptor (EPOR), EPO and Bcl-2 mRNAs were quantified by real time PCR. Activation of JAK-2, STAT-3 and STAT-5 in hepatocytes and rat livers perfused in situ was assessed by Western blotting.

Results

In contrast to previous in vivo studies on ischemia-reperfusion injury to the liver, rHu-EPO was without any protective effect on hypoxic injury, hypoxia-reoxygenation injury and cold-induced apoptosis to isolated cultured rat hepatocytes. EPOR mRNA was identified in these cells but specific detection of the EPO receptor protein was not possible due to the lack of antibody specificity. Both, in the cultured rat hepatocytes (10 U/ml for 15 minutes) and in the rat liver perfused in situ with rHu-EPO (8.9 U/ml for 15 minutes) no evidence for EPO-dependent signalling was found as indicated by missing effects of rHu-EPO on phosphorylation of JAK-2, STAT-3 and STAT-5 and on the induction of Bcl-2 mRNA.

Conclusion

Together, these results indicate the absence of any protective EPO signalling in rat hepatocytes. This implies that the protection provided by rHu-EPO in vivo against ischemia-reperfusion and other causes of liver injury is most likely indirect and does not result from a direct effect on hepatocytes.
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Metadata
Title
No evidence for protective erythropoietin alpha signalling in rat hepatocytes
Authors
Thorsten Bramey
Patricia Freitag
Joachim Fandrey
Ursula Rauen
Katja Pamp
Jochen Erhard
Stilla Frede
Herbert de Groot
Frank Petrat
Publication date
01-12-2009
Publisher
BioMed Central
Published in
BMC Gastroenterology / Issue 1/2009
Electronic ISSN: 1471-230X
DOI
https://doi.org/10.1186/1471-230X-9-26

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Keynote webinar | Spotlight on medication adherence

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WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

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