No Evidence for Protective Erythropoietin Alpha Signalling in Rat Hepatocytes

Overview

Journal BMC Gastroenterol

Publisher Biomed Central

Specialty Gastroenterology

Date 2009 Apr 23

PMID 19383129

Citations 5

Authors

Thorsten Bramey

Patricia Freitag

Joachim Fandrey

Ursula Rauen

Katja Pamp

Jochen Erhard

Stilla Frede

Herbert de Groot

Frank Petrat

Affiliations

Soon will be listed here.

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 degrees 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.

Citing Articles

Pharmacological Benefits and Risk of Using Hormones in Organ Perfusion and Preservation Solutions in the Aspect of Minimizing Hepatic Ischemia-Reperfusion Injury during Storage.

Ostrozka-Cieslik A, Dolinska B Biomed Res Int. 2019; 2019:6467134.

PMID: 31828112 PMC: 6881579. DOI: 10.1155/2019/6467134.

Hepatocellular heme oxygenase-1: a potential mechanism of erythropoietin-mediated protection after liver ischemia-reperfusion injury.

Riehle K, Hoagland V, Benz W, Campbell J, Liggitt D, Langdale L Shock. 2014; 42(5):424-31.

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The effect of erythropoietin on normal and neoplastic cells.

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Glycine, a simple physiological compound protecting by yet puzzling mechanism(s) against ischaemia-reperfusion injury: current knowledge.

Petrat F, Boengler K, Schulz R, de Groot H Br J Pharmacol. 2011; 165(7):2059-72.

PMID: 22044190 PMC: 3413844. DOI: 10.1111/j.1476-5381.2011.01711.x.

Effect of recombinant erythropoietin on ischemia-reperfusion-induced apoptosis in rat liver.

Shawky H, Younan S, Rashed L, Shoukry H J Physiol Biochem. 2011; 68(1):19-28.

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