Than Min Min, Koonyosying Pimpisid, Ruangsuriya Jetsada, Junrungsee Sunhawit, Uthaipibull Chairat, Srichairatanakool Somdet
Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
Department of Biochemistry, University of Medicine, Mandalay 05021, Myanmar.
Materials (Basel). 2021 Oct 28;14(21):6480. doi: 10.3390/ma14216480.
Iron is essential for all living organisms. It is strictly controlled by iron transporters, transferrin receptors, ferroportin and hepcidin. Erythroferrone (ERFE) is an iron-regulatory hormone which is highly expressed in erythroblasts by erythropoietin (EPO) stimulation and osteoblasts independently of EPO by sequestering bone morphogenetic proteins and inhibiting hepatic hepcidin expression. Although the hepcidin suppressive function of ERFE is known, its receptors still require investigation. Here, we aim to identify ERFE receptors on the HepG2 and Huh7 cells responsible for ERFE. Recombinant ERFE (rERFE) was first produced in HEK293 cells transfected with pcDNA3.1 + ERFE, then purified and detected by Western blot. The liver cells were treated with an rERFE-rich medium of transfected HEK293 cells and a purified rERFE-supplemented medium at various time points, and hepcidin gene () expression was determined using qRT-PCR. The results show that 37-kD rERFE was expressed in HEK293 cells. was suppressed at 3 h and 6 h in Huh7 cells after rERFE treatments ( < 0.05), then restored to the original levels. was activated after treatment with purified rERFE for 24 h and 48 h. Together, these results reveal that ERFE suppressed expression in liver cells, possibly acting on membrane ERFE receptor, which in Huh7 cells was more sensitive to the ERFE concentrate.
铁对所有生物都是必不可少的。它受到铁转运蛋白、转铁蛋白受体、铁转运蛋白和铁调素的严格调控。红细胞生成素铁（ERFE）是一种铁调节激素，在红细胞生成素（EPO）刺激下在成红细胞中高表达，在成骨细胞中则独立于EPO通过螯合骨形态发生蛋白并抑制肝脏铁调素表达而高表达。尽管已知ERFE具有抑制铁调素的功能，但其受体仍有待研究。在此，我们旨在鉴定负责ERFE的HepG2和Huh7细胞上的ERFE受体。重组ERFE（rERFE）首先在转染了pcDNA3.1 + ERFE的HEK293细胞中产生，然后纯化并通过蛋白质免疫印迹法检测。在不同时间点，用转染的HEK293细胞的富含rERFE培养基和添加了纯化rERFE的培养基处理肝细胞，并使用实时定量聚合酶链反应（qRT-PCR）测定铁调素基因（）表达量。结果显示，37-kD rERFE在HEK293细胞中表达了。rERFE处理后，Huh7细胞中的铁调素基因表达量在3小时和6小时受到抑制（P < 0.05），然后恢复到原始水平。用纯化的rERFE处理24小时和48小时后，铁调素基因表达量被激活。总之，这些结果表明ERFE抑制肝细胞中的铁调素基因表达，可能作用于膜ERFE受体，而在Huh7细胞中该受体对ERFE浓缩物更敏感。
