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 Blood, 15 June 2008, Vol. 111, No. 12, pp. 5727-5733.
Prepublished online as a Blood First Edition Paper on March 7, 2008; DOI 10.1182/blood-2007-08-106195.

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RED CELLS

Brief Report

Erythropoietin mediates hepcidin expression in hepatocytes through EPOR signaling and regulation of C/EBP{alpha}

Jorge P. Pinto1, Sara Ribeiro1,*, Helena Pontes2,*, Shifaan Thowfeequ3,*, David Tosh3, Félix Carvalho2, and Graça Porto1,4,5

1 Iron Genes and Immune System, Instituto de Biologia Molecular e Celular (IBMC), University of Porto, Porto, Portugal; 2 REQUIMTE (Rede de Química e Tecnologia), Toxicology Department, Faculty of Pharmacy, University of Porto, Porto, Portugal; 3 Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath, Bath, United Kingdom; 4 Clinical Hematology, Santo António General Hospital, Porto, Portugal; and 5 Molecular Immunology and Pathology, Abel Salazar Institute for the Biomedical Science (ICBAS), University of Porto, Porto, Portugal

Hepcidin is the principal iron regulatory hormone, controlling the systemic absorption and remobilization of iron from intracellular stores. Recent in vivo studies have shown that hepcidin is down-regulated by erythropoiesis, anemia, and hypoxia, which meets the need of iron input for erythrocyte production. Erythropoietin (EPO) is the primary signal that triggers erythropoiesis in anemic and hypoxic conditions. Therefore, a direct involvement of EPO in hepcidin regulation can be hypothesized. We report here the regulation of hepcidin expression by EPO, in a dose-dependent manner, in freshly isolated mouse hepatocytes and in the HepG2 human hepatocyte cell model. The effect is mediated through EPOR signaling, since hepcidin mRNA levels are restored by pretreatment with an EPOR-blocking antibody. The transcription factor C/EBP{alpha} showed a pattern of expression similar to hepcidin, at the mRNA and protein levels, following EPO and anti-EPOR treatments. Chromatin immunoprecipitation experiments showed a significant decrease of C/EBP{alpha} binding to the hepcidin promoter after EPO supplementation, suggesting the involvement of this transcription factor in the transcriptional response of hepcidin to EPO.


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