REGULATION OF HEPCIDIN BY ERYTHROPOIETIN IN MACROPHAGES

IOLSTER J; VITTORI D; NESSE A; MALTANERI R; CHAMORRO ME

Congreso; LXVI Reunión Científica de la Sociedad Argentina de Investigación Clínica; 2021

Sociedad Argentina de Investigación Clínica

The role of erythropoietin (Epo) as a growth factor in erythropoiesis is well known. Many defects, both congenital and acquired, lead to ineffective erythropoiesis, thus establishing the condition of anemia. The demand for iron (Fe), necessary in biological processes, including erythropoiesis, gives importance to its controlled regulation. Hepcidin (Hep) is a key regulator of systemic iron metabolism and its expression responds to Fe levels. The participation of Epo in the regulation of Hep has been investigated in hepatocytes but not in macrophages, fundamental cells in Fe homeostasis that act as reservoirs of senescent erythrocytes. Therefore, the aim of this work was to investigate the regulation of Hep by Epo in macrophages.Standardization of the differentiation of monocytic THP-1 cells to macrophages was carried out with different concentrations of phorbol 12-myristate-13-acetate (PMA). According to cell morphology (visible light microscopy), results of viability, proliferation and mRNA levels of the specific differentiation markers CD-14 and CD-68 (RT-PCR), 100 nM was chosen as the adequate PMA concentration to continue with the stated objective. The presence of EpoR was demonstrated by Western blotting (anti-EpoR M20), RT-PCR and flow cytometry, comparing with UT-7 cells as a positive control. Epo treatment of macrophages induced a significant decrease in Hep mRNA levels (RT-PCR, a.u.: C6h 0.39±0.02; *Epo6h 0.11±0.03; *Epo24h 0.09±0.04, *P