Guinea pig intestinal iron binding protein

An iron binding protein, isolated from guinea pig intestinal mucosa, was compared to guinea pig transferrin. Both had a molecular weight of approximately 80,000. The intestinal iron-binding protein consisted of 2 subunits of equal molecular weight; transferrin had no subunits. Transferrin showed an absorbance peak at 470 nm; the intestinal iron-binding protein had no visible absorbance but did have a peak at 336 nm. Electron spin resonance spectra of the two proteins dfffered. Significant differences on amino acid analysis were also identified.     

Molecular mechanism of intestinal iron absorption

Iron is an essential trace metal in the human diet because of its role in a number of metabolic processes including oxygen transport. In the diet, iron is present in two fundamental forms, heme and non-heme iron. This article presents a brief overview of the molecular mechanisms of intestinal iron absorption and its regulation. While many proteins that orchestrate iron transport pathway have been identified, a number of key factors that control the regulation of iron absorption still remain to be elucidated. This review also summarizes new and emerging information about iron metabolic regulators that coordinate regulation of intestinal iron absorption.     

Systemic regulation of intestinal iron absorption

The intestinal absorption of the essential trace element iron and its mobilization from storage sites in the body are controlled by systemic signals that reflect tissue iron requirements. Recent advances have indicated that the liver-derived peptide hepcidin plays a central role in this process by repressing iron release from intestinal enterocytes, macrophages and other body cells. When iron requirements are increased, hepcidin levels decline and more iron enters the plasma. It has been proposed that the level of circulating diferric transferrin, which reflects tissue iron levels, acts as a signal to alter hepcidin expression. In the liver, the proteins HFE, transferrin receptor 2 and hemojuvelin may be involved in mediating this signal as disruption of each of these molecules decreases hepcidin expression. Patients carrying mutations in these molecules or in hepcidin itself develop systemic iron loading (or hemochromatosis) due to their inability to down regulate iron absorption. Hepcidin is also responsible for the decreased plasma iron or hypoferremia that accompanies inflammation and various chronic diseases as its expression is stimulated by pro-inflammatory cytokines such as interleukin 6. The mechanisms underlying the regulation of hepcidin expression and how it acts on cells to control iron release are key areas of ongoing research.     

Toxicological effects of iron on intestinal cells

The aim of the present study was to investigate whether iron, which is involved in the formation of free radicals in vitro, can initiate cellular injury in human intestinal cells. The effects of various concentrations of iron were studied in preconfluent, colonic-cancerogenous cells, and also in postconfluent, differentiating cells. Cellular damage was assessed using cell proliferation (serial cell counting), tetrazolium dye (MTT) uptake, lactate dehydrogenase (LDH) release and apoptosis studies based on caspase-3 activities. Also the activities of the major antioxidative enzymes, superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) were measured after the cells had been exposed to iron. Our results indicated that preconfluent cells were more susceptible to iron toxicity, as assessed by a significant reduction in cell proliferation and MTT uptake in a concentration-dependent manner compared to the control. However, no evidence for MTT uptake was observed in postconfluent cells. Caspase-3 activity, an indicator of cell apoptosis, considerably increased in preconfluent cells at high iron levels compared to the control (p < 0.05), whereas postconfluent cells were not significantly affected. LDH release was similar for both groups and was significantly higher than the control at 900 microM iron and above. SOD activities were not affected by iron in either group, whereas GPx was considerably higher in iron-treated cells in both groups compared with the control (because of relatively high standard deviations this effect was not significant). In conclusion we suggest that iron exerts its toxic effects intracellularly especially in preconfluent Caco-2 cells, whereas only high iron doses were able to alter the viability of differentiating, enterocyte-like cells.     

Effect of zinc depletion/repletion on intestinal iron absorption and iron status in rats

Iron and zinc deficiencies likely coexist in general population. We have previously demonstrated that zinc treatment induces while zinc deficiency inhibits iron absorption in intestinal cell culture models, but this needs to be tested in vivo. In the present study we assessed intestinal iron absorption, iron status (haemoglobin), red blood cell number, plasma ferritin, transferrin receptor, hepcidin) and tissue iron levels in zinc depleted, replete and pair fed control rats. Zinc depletion led to reduction in body weight, tissue zinc levels, intestinal iron absorption, protein and mRNA expression of iron transporters, the divalent metal ion transporter-1, hephaestin and ferroportin, but elevated the intestinal and liver tissue iron levels compared with the pair fed control rats. Zinc repletion led to a significant weight gain compared to zinc deficient rats and normalized the iron absorption, iron transporter expression, tissue iron levels to that of pair fed control rats. Surprisingly, haemoglobin levels and red blood cell number reduced significantly in zinc repleted rats, which could be due to rapid weight gain. Together, these results indicate that whole body zinc status has profound influence on growth, intestinal absorption and systemic utilization of iron, mediated via modulation of iron transporter expression.     

Unexpected role of ceruloplasmin in intestinal iron absorption

Ferroxidases are essential for normal iron homeostasis in most organisms. The paralogous vertebrate ferroxidases ceruloplasmin (Cp) and hephaestin (Heph) are considered to have nonidentical functions in iron transport: plasma Cp drives iron transport from tissue stores while intestinal Heph facilitates iron absorption from the intestinal lumen. To clarify the function of Cp, we acutely bled Cp-/- mice to stress iron homeostasis pathways. Red cell hemoglobin recovery was defective in stressed Cp-/- mice, consistent with low iron availability. Contrary to expectations, iron was freely released from spleen and liver stores in Cp-/- mice, but intestinal iron absorption was markedly impaired. Phlebotomy of wild-type mice caused a striking shift of Cp from the duodenal epithelium to the underlying lamina propria, suggesting a critical function of Cp in basolateral iron transport. Regulated relocalization of intestinal Cp may represent a fail-safe mechanism in which Cp shares with Heph responsibility for iron absorption under stress.     

Aerobactin-mediated iron uptake system in intestinal Escherichia coli

Abstract Escherichia coli strains isolated from children with diarrhea were studied to determine the incidence of the components of the aerobactin system. The production of aerobactin and the presence of the aerobactin receptor were demonstrated using bioassay techniques. The presence and location of the genes for the aerobactin system was determined by colony and Southern DNA-DNA hydridization. It was found that aerobactin and the aerobactin receptor could be coded by either chromosomal or plasmid genes. A high percentage of strains expressed the receptor in the absence of aerobactin. Of 19 enterotoxin-producing strains one was found to have the complete aerobactin system, while 11 expressed only the aerobactin receptor which was most frequently coded by a plasmid.     

Studies on the role of iron binding ligands and the intestinal brush border receptors in iron absorption.

With a view to identifying ligands that could be used as promoters of iron absorption, the affinity of a number of iron chelating agents and the efficiency with which they can donate iron to the brush border receptors has been studied. A number of organic and inorganic compounds were found to chelate iron and keep it soluble at pH 7.5 of the intestinal lumen. This ligand-bound iron was taken up by the intestinal brush border receptors with varying degree of efficiency; ascorbic acid being the most effective and EDTA and citrate the least effective in donating the chelated iron to the receptors. Several polyphosphate compounds, used as food additives, chelated iron and kept it in solution but showed moderate potency for donating iron to the receptors.     

缺铁性贫血中肠道菌群作用的研究进展

缺铁性贫血是临床最常见的贫血类型之一,已知缺铁性贫血会导致机体免疫力下降,影响心血管系统,对儿童和青少年的智力发育造成损害。然而越来越多的学者发现缺铁还会引起肠道菌群结构发生改变,影响肠道菌群的代谢。短链脂肪酸作为肠道菌群代谢的主要产物,受铁的制约最大。短链脂肪酸对机体肠道健康起着至关重要的作用,因此缺铁是有害的。另外临床工作中发现益生菌联合铁剂治疗缺铁性贫血,疗效更显著；铁剂联合抗菌药物对细菌相关疾病的治疗效果大大提高,说明铁与肠道菌群关系密切。本文就缺铁性贫血与肠道菌群关系的研究进展作一综述。     

Iron Imports. VI. HFE and regulation of intestinal iron absorption

The majority of clinical cases of iron overload is caused by mutations in the HFE gene. However, the role that HFE plays in the physiology of intestinal iron absorption remains enigmatic. Two major models have been proposed: 1) HFE exerts its effects on iron homeostasis indirectly, by modulating the expression of hepcidin; and 2) HFE exerts its effects directly, by changing the iron status (and therefore the iron absorptive activity) of intestinal enterocytes. The first model places the primary role of HFE in the liver (hepatocytes and/or Kupffer cells). The second model places the primary role in the duodenum (crypt cells or villus enterocytes). These models are not mutually exclusive, and it is possible that HFE influences the iron status in each of these cell populations, leading to cell type-specific downstream effects on intestinal iron absorption and body iron distribution.     

Ultrastructural silver enhancement of Prussian blue-reactive iron in hematopoietic and intestinal cells

Prussian blue has been widely used to localize iron in a variety of tissues at the light and electron microscopic level. In the present study, thin sections of human marrow and blood cells and rat duodenal cells were exposed to silver proteinate (SP) after staining en bloc with acid ferrocyanide (AF), with and without prior iron saturation using iron nitrilotriacetate (FeNTA). Silver deposition was observed over Prussian blue-reactive sites and significantly enhanced sites of minimal AF and FeNTA-AF staining. AF-SP stain deposits were present in the cytoplasmic matrix, granules, and occasionally on the surfaces of macrophages, monocytes, and erythroblasts. FeNTA-AF-SP stained additional cytoplasmic and surface sites in erythroblasts and stained neutrophil granules intensely. Duodenal epithelium from iron-loaded rats demonstrated strong AF-SP staining of ferric iron in microvilli, apical cytoplasmic matrix, and lateral membranes. Similar preparations from iron-replete rats stained sparsely; however, intense AF-SP staining was observed after iron saturation with FeNTA. SP similarly enhanced luminal ferrous iron deposits stained with acid ferricyanide in rats given intraluminal ferrous iron. AF-SP stain deposits were removed by exposure of thin sections to NH4OH, KCN, or HNO3 but were not affected by prior exposure to HIO4 or NaBH4, consistent with a silver cyanide or complex stain precipitate rather than reduced silver or silver ferriferrocyanide. SP enhancement of Prussian blue allows identification of reactive sites not readily visualized with AF or FeNTA-AF alone, and offers the potential for differentiating AF staining from other deposits or organelles of comparable density.     

Transferrin stimulates iron absorption, exocytosis, and secretion in cultured intestinal cells

The cellularmechanism by which basolateral transferrin (Tf) produces an increase inapical-to-basolateral Fe flux in Caco-2 cells was analyzed. After apulse of ⁵⁹Fe from the apicalmedium, three types of basolateral⁵⁹Fe efflux were found: a⁵⁹Fe efflux that was independentof the presence of Tf in the basolateral medium, a⁵⁹Fe efflux in which⁵⁹Fe left the cell bound to Tf,and a Tf-dependent ⁵⁹Fe efflux inwhich ⁵⁹Fe came off the cell notbound to Tf. Furthermore, addition of Tf to the basolateral mediumdoubled the exocytosis rate of Tf and increased the secretion ofapolipoprotein A, a basolateral secretion marker. Both apotransferrinand Fe-containing Tf produced similar increases in⁵⁹Fe efflux, Tf exocytosis, andapolipoprotein A secretion. The Ca²⁺ channel inhibitor SKF-96365inhibited both the Tf-mediated increase in transepithelial Fe transportand the secretion of apolipoprotein A. Thus the activation oftransepithelial Fe transport by Tf seems to be mediated byCa²⁺ entry into the cells.     

Tumour necrosis factor alpha causes hypoferraemia and reduced intestinal iron absorption in mice

Cytokines are implicated in the anaemia of chronic disease by reducing erythropoiesis and increasing iron sequestration in the reticuloendotheial system. However, the effect of cytokines, in particular TNFalpha (tumour necrosis factor alpha), on small bowel iron uptake and iron-transporter expression remains unclear. In the present study, we subjected CD1 male mice to intraperitoneal injection with TNFalpha (10 ng/mouse) and then examined the expression and localization of DMT1 (divalent metal transporter 1), IREG1 (iron-regulated protein 1) and ferritin in duodenum. Liver and spleen samples were used to determine hepcidin mRNA expression. Changes in serum iron and iron loading of duodenum, spleen and liver were also determined. We found a significant (P<0.05) fall in serum iron 3 h post-TNFalpha exposure. This was coincident with increased iron deposition in the spleen. After 24 h of exposure, there was a significant decrease in duodenal iron transfer (P<0.05) coincident with increased enterocyte ferritin expression (P<0.05) and re-localization of IREG1 from the basolateral enterocyte membrane. Hepatic hepcidin mRNA levels remained unchanged, whereas splenic hepcidin mRNA expression was reduced at 24 h. In conclusion, we provide evidence that TNFalpha may contribute to anaemia of chronic disease by iron sequestration in the spleen and by reduced duodenal iron transfer, which seems to be due to increased enterocyte iron binding by ferritin and a loss of IREG1 function. These observations were independent of hepcidin mRNA levels.     

Effect of orally-administered epidermal growth factor on intestinal iron absorption and mucosal permeability

A progressive increase in intestinal 59Fe3+ absorption was observed on oral feeding of mice with physiological doses of EGF/UGO. Maximal changes were apparent after 3d and appeared to be dose-dependent. In addition to a small increase in intestinal cell proliferation, as reflected by increased ornithine decarboxylase activity, EGF/UGO-feeding increased mucosal permeability (evaluated with [51Cr]-EDTA): the latter could account for the increase in iron absorption. Sialoadenectomy, to remove the major source of endogenous EGF/UGO, had no appreciable effect on the intestinal absorption of iron.     

Increased intestinal iron absorption in rats with normal hepatic iron stores. Kinetic aspects of the adaptative response to parenteral iron repletion in dietary iron deficiency

Male Sprague-Dawley rats were fed an iron-deficient diet for 8 days. After this period, iron stores were repleted in three groups of animals by intravenous administration of iron dextran. In a second set of experiments, iron was administered in the same dose as Fe nitrilotriacetic acid complex. 12 h, 24 h and 48 h thereafter, the intestinal iron transfer in vitro and in vivo as well as the non-heme iron and ferritin content were determined in both the liver and the jejunal mucosa. In iron deficiency, intestinal iron transfer is increased to 230% of untreated controls, while non-heme iron and ferritin decreased to 20% and 10% in the liver and to 55% and 25% in the mucosa, respectively. 12 h and 24 h after parenteral administration of 0.1 mmol Fe/kg body weight iron transfer was as high as in iron deficiency, while liver iron stores were not significantly different from the untreated controls. In this situation, the close link between decreases in body iron stores and increases in iron transfer was temporarily dissociated. This can be related to the time lag between the incorporation of parenterally applied iron in the liver and in the jejunal mucosa. The data provide evidence for the hypothesis that the hepatic iron stores have no means of neural or hormonal communication with the small intestine in order to adapt iron transfer to their state of repletion on short notice. Intestinal iron transfer returned to control levels after 48 h.