Knutson Mitchell D, Vafa Mohammad R, Haile David J, Wessling-Resnick Marianne
Harvard School of Public Health, Dept of Nutrition, Bldg 2, Rm 205, 665 Huntington Ave, Boston, MA 02115, USA.
Blood. 2003 Dec 1;102(12):4191-7. doi: 10.1182/blood-2003-04-1250. Epub 2003 Aug 7.
The expression of ferroportin1 (FPN1) in reticuloendothelial macrophages supports the hypothesis that this iron-export protein participates in iron recycling from senescent erythrocytes. To gain insight into FPN1's role in macrophage iron metabolism, we examined the effect of iron status and erythrophagocytosis on FPN1 expression in J774 macrophages. Northern analysis indicated that FPN1 mRNA levels decreased with iron depletion and increased on iron loading. The iron-induced induction of FPN1 mRNA was blocked by actinomycin D, suggesting that transcriptional control was responsible for this effect. After erythrophagocytosis, FPN1 mRNA levels were also up-regulated, increasing 8-fold after 4 hours and returning to basal levels by 16 hours. Western analysis indicated corresponding increases in FPN1 protein levels, with maximal induction after 10 hours. Iron chelation suppressed FPN1 mRNA and protein induction after erythrophagocytosis, suggesting that FPN1 induction results from erythrocyte-derived iron. Comparative Northern analyses of iron-related genes after erythrophagocytosis revealed a 16-fold increase in FPN1 levels after 3 hours, a 10-fold increase in heme oxygenase-1 (HO-1) after 3 hours, a 2-fold increase in natural resistance macrophage-associated protein 1 (Nramp1) levels after 6 hours, but no change in divalent metal ion transporter 1 (DMT1) levels. The rapid and strong induction of FPN1 expression after erythrophagocytosis suggests that FPN1 plays a role in iron recycling.
网织内皮巨噬细胞中铁转运蛋白1(FPN1)的表达支持了这样一种假说,即这种铁输出蛋白参与衰老红细胞中铁的再循环。为深入了解FPN1在巨噬细胞铁代谢中的作用,我们检测了铁状态和红细胞吞噬作用对J774巨噬细胞中FPN1表达的影响。Northern印迹分析表明,FPN1 mRNA水平在铁缺乏时降低,在铁负载时升高。放线菌素D可阻断铁诱导的FPN1 mRNA的上调,提示转录调控介导了这一效应。红细胞吞噬作用后,FPN1 mRNA水平也上调,4小时后增加8倍,16小时后恢复至基础水平。Western印迹分析表明FPN1蛋白水平相应增加,10小时后诱导作用最强。铁螯合剂抑制红细胞吞噬作用后FPN1 mRNA和蛋白的诱导,提示FPN1的诱导源于红细胞来源的铁。红细胞吞噬作用后对铁相关基因的比较Northern印迹分析显示,3小时后FPN1水平增加16倍,3小时后血红素加氧酶-1(HO-1)增加10倍,6小时后天然抗性巨噬细胞相关蛋白1(Nramp1)水平增加2倍,但二价金属离子转运体1(DMT1)水平无变化。红细胞吞噬作用后FPN1表达的快速且强烈诱导提示FPN1在铁再循环中发挥作用。