铁调素对小鼠单核细胞RAW264.7膜铁转运蛋白1(FPN1)表达的影响

Effect of hepcidin on the expression of ferroportin 1 in mouse RAW264.7 cells

目的观察铁调素对小鼠单核细胞RAW264.7膜铁转运蛋白1(FPN1)的表达,探讨铁调素对RAW264.7细胞作用的可能通路和机制。方法将不同浓度的铁调素加入含有核因子κB受体活化因子配体(RANKL)的RAW264.7细胞培养基,24h后用免疫荧光和Western-blot方法测定FPN1的表达,共聚焦显微镜(CLSM)测定细胞内铁离子的浓度。结果 RAW264.7细胞膜上存在FPN1受体的阳性表达;在本实验铁调素浓度干预范围内,FPN1的表达随着铁调素浓度的增加呈浓度依赖性降低(P<0.05),同时细胞内铁离子的含量随着铁调素浓度的增加呈浓度依赖性增加(P<0.05)。结论小鼠单核细胞RAW264.7是铁调素作用的靶细胞,铁调素可通过降解其细胞膜上的FPN1增加细胞内的铁离子。

Objective To investigate the effect of hepcidin on the expression of ferroportin 1 （ FPN1 ） in RAW264.7 cells in vitro, and to explore the possible mechanism.Methods Mouse monocytes RAW264.7 were treated with different concentrations of hepcidin in the presence of receptor activator of NF-Kb ligand （ RANKL） .After 24 hours, the expression of FPN1 was detected using Western blotting and immunofluorescence, respectively.The intracellular iron concentration was measured using a confocal laser scanning microscope.Results The results documented the existence of FPN1 at the membrane of RAW264.7 cells, and the expression of FPN1 was markedly downoregulated by hepcidin in a concentration-dependent manner （ p 〈0.05 ） .Furthermore, hepcidin increased intracellular iron in RAW264.7 cells in a concentration-dependent manner （p〈0.05）.Conclusion RAW264.7 cells are target cells of hepcidin.Hepcidin can increase intracellular iron by degradation of FPN1 at the membrane of RAW264.7 cells.