Medical Student USF Health Morsani College of Medicine Brandon, Florida, United States
Background: Premature birth predisposes infants to iron deficiency due to interrupted maternal fetal iron transfer. In adults, hepcidin inhibits intestinal iron absorption via Ferroportin (FPN) and Divalent Metal Transporter 1 (DMT1) proteasomal protein degradation. This process is unknown in preterm infants. A better understanding of the regulatory role of hepcidin on iron absorption in immature intestine will help to improve the care of preterm infants. Objective: The purpose of this study is to determine the effects of hepcidin on FPN and DMT1 in vitro immature intestinal epithelial cells. Design/Methods: Fetal tissue-derived enteroids were established with both basolateral and apical out orientations. The basolateral-out enteroids were exposed to a low concentration of hepcidin at 20 µg/L in the growth media. Three repeated experiments were conducted, each consisted of hepcidin and a negative control (no exposure) in triplicates. RT-qPCR was performed to quantify gene expression of FPN and DMT1. We will test the effect of hepcidin on DMT1 with the apical out enteroids to ensure direct contact. In addition to quantifying gene expression, we will perform Western blot to compare the protein productions through signal intensity of the band. The enteroids will be exposed to a range of hepcidin concentrations equivalent to serum concentrations to determine the dose dependent effects. The Pfaffl method will be used to estimate gene expression and means of gene expression will be compared using student t-test. These experiments will be completed by March 2024.
