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Biochemical and Molecular Actions of Nutrients

Duodenal Ascorbate Levels Are Changed in Mice with Altered Iron Metabolism^1,2

Bisera Atanasova^*, Ian S. Mudway^,**, Abas H. Laftah^**, Gladys O. Latunde-Dada^**, Andrew T. Mckie^**, Timothy J. Peters, Kamen N. Tzatchev^* and Robert J. Simpson^**,3

^* Department of Clinical Laboratory and Clinical Immunology, Medical University, Sofia, Bulgaria; Lung Biology, Department of Health and ^** Department of Life Sciences, King’s College London, UK; and Department of Clinical Biochemistry, GKT School of Medicine, King’s College, Denmark Hill Campus, London, UK

³To whom correspondence should be addressed. E-mail: robert.simpson{at}kcl.ac.uk.

Ascorbate has long been thought to play an important role in intestinal iron absorption. The recent identification of a possible ascorbate-dependent duodenal ferric reductase suggests a role for intracellular ascorbate in the control of iron absorption. We set out to determine whether duodenal ascorbate concentrations are altered by treatments known to alter the rate of iron absorption and whether ascorbate levels affect duodenal reductase activity. Duodenal ascorbate was extracted and assayed by HPLC and/or a chemical assay. Ferric reductase was assayed in vitro with ferric nitrilotriacetate or nitroblue tetrazolium as substrates. Duodenal ascorbate concentrations were increased by iron deficiency, genetic hypotransferrinemia, and hypoxia. Parenteral iron overload increased iron stores but did not affect duodenal ascorbate concentrations. Hemolytic anemia induced in mice by phenylhydrazine injection also did not affect duodenal ascorbate concentrations. In vitro studies with incubated duodenum showed that decreased tissue ascorbate was associated with decreased mucosal ferric reductase activity, whereas incubation with dehydroascorbate prevented both the decrease in ascorbate concentration and reductase activity. Mouse duodenum ascorbate concentrations changed in response to treatments that altered iron absorption rates; in particular, ascorbate levels generally increased when iron absorption was increased by iron deficiency, hypoxia, or genetic hypotransferrinemia. We conclude that changes in ascorbate levels are associated with changes in ferric reductase activity. These findings are consistent with the proposal that duodenal ascorbate plays a role in intestinal iron absorption.

KEY WORDS: • iron absorption • ferric reductase • iron overload • iron deficiency • hypotransferrinemia

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