Ferroportin mediates the intestinal absorption of iron from a nanoparticulate ferritin core mimetic in mice

Mohamad F Aslam, David M Frazer, Nuno Faria, Sylvaine F A Bruggraber, Sarah J Wilkins, Cornel Mirciov, Jonathan J Powell, Greg J Anderson, Dora I A Pereira, Mohamad F Aslam, David M Frazer, Nuno Faria, Sylvaine F A Bruggraber, Sarah J Wilkins, Cornel Mirciov, Jonathan J Powell, Greg J Anderson, Dora I A Pereira

Abstract

The ferritin core is composed of fine nanoparticulate Fe(3+) oxohydroxide, and we have developed a synthetic mimetic, nanoparticulate Fe(3+) polyoxohydroxide (nanoFe(3+)). The aim of this study was to determine how dietary iron derived in this fashion is absorbed in the duodenum. Following a 4 wk run-in on an Fe-deficient diet, mice with intestinal-specific disruption of the Fpn-1 gene (Fpn-KO), or littermate wild-type (WT) controls, were supplemented with Fe(2+) sulfate (FeSO4), nanoFe(3+), or no added Fe for a further 4 wk. A control group was Fe sufficient throughout. Direct intestinal absorption of nanoFe(3+) was investigated using isolated duodenal loops. Our data show that FeSO4 and nanoFe(3+) are equally bioavailable in WT mice, and at wk 8 the mean ± SEM hemoglobin increase was 18 ± 7 g/L in the FeSO4 group and 30 ± 5 g/L in the nanoFe(3+) group. Oral iron failed to be utilized by Fpn-KO mice and was retained in enterocytes, irrespective of the iron source. In summary, although nanoFe(3+) is taken up directly by the duodenum its homeostasis is under the normal regulatory control of dietary iron absorption, namely via ferroportin-dependent efflux from enterocytes, and thus offers potential as a novel oral iron supplement.

Keywords: basolateral export; hepcidin; iron homeostasis; knockout mice; nanoiron.

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Figures

Figure 1.
Figure 1.
Hb levels in mice during dietary iron repletion. Hb values for WT (solid trace) and Fpn KO (dashed trace) mice in the control Fe-sufficient (i.e., non-iron-depleted) group (A), the Fe-deficient group (B), the FeSO4-supplemented group (C), and the nanoFe3+-supplemented group (D) after iron depletion (study wk 4), 1 wk into the iron repletion period (study wk 5), 2 wk into the iron repletion period (study wk 6), and at the end of the iron repletion period (study wk 8) as per study outline presented in Supplemental Fig. S2. Arrows (C, D) indicate the start of iron repletion with FeSO4- or nanoFe3+-supplemented diets, respectively. Values are means ± sd. Numbers in each group are as follows: Fe-sufficient, n = 6 WT and n = 8 Fpn KO; Fe-deficient, n = 6 WT and n = 6 Fpn KO; FeSO4, n = 4 WT and n = 9 Fpn KO; nanoFe3+, n = 8 WT and n = 6 Fpn KO. *P ≤ 0.01, **P ≤ 0.007, ***P ≤ 0.0002, ****P < 0.0001 vs. Fpn-KO mice.
Figure 2.
Figure 2.
Tissue Fe distribution. Representative images of Perls' Prussian blue staining of duodenum and spleen of WT and Fpn-KO mice in the Fe-deficient (A), control Fe-sufficient (B), FeSO4-supplemented (C), or nanoFe3+-supplemented (D) groups, defined as per study outline presented in Supplemental Fig. S2. Arrows indicate example locations of iron staining in the tissues. Scale bars = 30 μm.
Figure 3.
Figure 3.
Liver and spleen nonheme iron levels. Hepatic iron (A) and splenic iron (B) levels of WT (solid bars) and Fpn-KO (open bars) mice of the control Fe-sufficient (i.e., non-iron-depleted) group, Fe-deficient group, FeSO4-supplemented group, and nanoFe3+-supplemented group. Data represent means ± sd. Numbers in each group are as follows: Fe-sufficient, n = 6 WT and n = 8 Fpn KO; Fe-deficient, n = 6 WT and n = 6 Fpn KO; FeSO4, n = 4 WT and n = 9 Fpn KO; nanoFe3+, n = 8 WT and n = 6 Fpn KO. *P ≤ 0.04, **P ≤ 0.006, ***P ≤ 0.0009, ****P < 0.0001.
Figure 4.
Figure 4.
Expression of Slc11a2 mRNA (A) in isolated enterocytes and Hamp1 (B) in the liver. Data shown for WT (solid bars) and Fpn-KO (open bars) mice in the control Fe-sufficient (i.e., non-iron-depleted) group, Fe-deficient group, FeSO4-supplemented group, and nanoFe3+-supplemented group. Values are means ± sd of the log2-transformed gene expression in relation to the housekeeping gene HPRT. Numbers in each group are as follows: Fe-sufficient, n = 6 WT and n = 8 Fpn KO; Fe-deficient, n = 6 WT and n = 6 Fpn KO; FeSO4, n = 4 WT and n = 9 Fpn KO; nanoFe3+, n = 8 WT and n = 6 Fpn KO. **P ≤ 0.007, ***P ≤ 0.0006, ****P < 0.0001.
Figure 5.
Figure 5.
Serum iron levels in Fpn-KO (open bars) and WT (solid bars) mice following 30 min exposure of duodenal loops to iron. Duodenal loops in male mice (8–12 wk old) were infused with either saline (control), 500 μM iron as FeNTA2, or 500 μM iron as nanoFe3+. Data represent means ± sd. Numbers in each group are as follows: saline control, n = 6 WT and n = 7 Fpn KO; FeNTA2, n = 7 WT and n = 7 Fpn KO; nanoFe3+, n = 10 WT and n = 9 Fpn KO. *P ≤ 0.04, **P = 0.007.

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