Erythropoietin mediates hepcidin expression in hepatocytes through EPOR signaling and regulation of C/EBPalpha.
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/EBPalpha 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/EBPalpha binding to the hepcidin promoter after EPO supplementation, suggesting the involvement of this transcription factor in the transcriptional response of hepcidin to EPO.
Pinto
Jorge P
JP
Iron Genes and Immune System, Instituto de Biologia Molecular e Celular, University of Porto, Porto, Portugal.
Ribeiro
Sara
S
Pontes
Helena
H
Thowfeequ
Shifaan
S
Tosh
David
D
Carvalho
Félix
F
Porto
Graça
G
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000235:genetics*; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004920:Erythropoiesis / Q000502:physiology; D004921:Erythropoietin / Q000378:metabolism* / Q000494:pharmacology; D015870:Gene Expression / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology*; D064451:Hepcidins; D006801:Humans; D008113:Liver Neoplasms; D051379:Mice; D008810:Mice, Inbred C57BL; D011401:Promoter Regions, Genetic / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D017467:Receptors, Erythropoietin / Q000276:immunology / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
D000906:Antibodies; D023181:Antimicrobial Cationic Peptides; D022763:CCAAT-Enhancer-Binding Protein-alpha; C578702:HAMP protein, human; C579599:Hamp protein, mouse; C474320:Hamp2 protein, mouse; D064451:Hepcidins; D012333:RNA, Messenger; D017467:Receptors, Erythropoietin; D004921:Erythropoietin