Nutr Res Pract.  2015 Dec;9(6):613-618. 10.4162/nrp.2015.9.6.613.

Effects of developmental iron deficiency and post-weaning iron repletion on the levels of iron transporter proteins in rats

• Oh S
• Shin PK
• Chung J

• KMID: 2313885
• DOI: http://doi.org/10.4162/nrp.2015.9.6.613

Abstract

BACKGROUND/OBJECTIVES
Iron deficiency in early life is associated with developmental problems, which may persist until later in life. The question of whether iron repletion after developmental iron deficiency could restore iron homeostasis is not well characterized. In the present study, we investigated the changes of iron transporters after iron depletion during the gestational-neonatal period and iron repletion during the post-weaning period.
MATERIALS/METHODS
Pregnant rats were provided iron-deficient (< 6 ppm Fe) or control (36 ppm Fe) diets from gestational day 2. At weaning, pups from iron-deficient dams were fed either iron-deficient (ID group) or control (IDR group) diets for 4 week. Pups from control dams were continued to be fed with the control diet throughout the study period (CON).
RESULTS
Compared to the CON, ID rats had significantly lower hemoglobin and hematocrits in the blood and significantly lower tissue iron in the liver and spleen. Hepatic hepcidin and BMP6 mRNA levels were also strongly down-regulated in the ID group. Developmental iron deficiency significantly increased iron transporters divalent metal transporter 1 (DMT1) and ferroportin (FPN) in the duodenum, but decreased DMT1 in the liver. Dietary iron repletion restored the levels of hemoglobin and hematocrit to a normal range, but the tissue iron levels and hepatic hepcidin mRNA levels were significantly lower than those in the CON group. Both FPN and DMT1 protein levels in the liver and in the duodenum were not different between the IDR and the CON. By contrast, DMT1 in the spleen was significantly lower in the IDR, compared to the CON. The splenic FPN was also decreased in the IDR more than in the CON, although the difference did not reach statistical significance.
CONCLUSIONS
Our findings demonstrate that iron transporter proteins in the duodenum, liver and spleen are differentially regulated during developmental iron deficiency. Also, post-weaning iron repletion efficiently restores iron transporters in the duodenum and the liver but not in the spleen, which suggests that early-life iron deficiency may cause long term abnormalities in iron recycling from the spleen.

Keyword

Developmental iron deficiency; ferroportin; divalent metal transporter 1; rat

MeSH Terms

Animals
Diet
Duodenum
Hematocrit
Hepcidins
Homeostasis
Iron*
Iron, Dietary
Liver
Rats*
Recycling
Reference Values
RNA, Messenger
Spleen
Weaning
Iron
Iron, Dietary
RNA, Messenger

Figure

• 

  Fig. 1 Effects of developmental iron deficiency and the post-weaning iron repletion on the protein levels of ferritin and transferrin receptor (TfR) in the liver (A) and spleen (B). CON: control diet, ID: iron-deficient diet, IDR: iron-deficient diet followed by control diet. Upper panels: representative blots. Lower panels: Densitometric analyses. Data are means ± SEM, n = 6-8/group. Different superscript means significant difference, P < 0.05.

• 

  Fig. 2 Effects of developmental iron deficiency and the post-weaning iron repletion on the mRNA levels of hepcidin (A) and BMP6 (B). CON: control diet, ID: iron-deficient diet, IDR: iron-deficient diet followed by control diet. β-actin was used as the housekeeping gene. Data are means ± SEM, n = 6-8/group. Different superscript means significant difference, P < 0.05.

• 

  Fig. 3 Effects of developmental iron deficiency and the post-weaning iron repletion on the protein levels of FPN and DMT1 expression in the duodenum (A), liver (B), and spleen (C). CON: control diet, ID: iron-deficient diet, IDR: iron-deficient diet followed by control diet. Upper panels: representative blots. Lower panels: Densitometric analyses. Data are means ± SEM, n = 6-8/group. Different superscript means significant difference, P < 0.05.

Reference

1. Brigham D, Beard J, Tobin B. Iron and thermoregulation: a review. Crit Rev Food Sci Nutr. 1996; 36:747–763.

2. Lozoff B, Brittenham GM. Behavioral aspects of iron deficiency. Prog Hematol. 1986; 14:23–53.

3. Falkingham M, Abdelhamid A, Curtis P, Fairweather-Tait S, Dye L, Hooper L. The effects of oral iron supplementation on cognition in older children and adults: a systematic review and meta-analysis. Nutr J. 2010; 9:4.

4. Lozoff B. Early iron deficiency has brain and behavior effects consistent with dopaminergic dysfunction. J Nutr. 2011; 141:740S–746S.

5. Greminger AR, Mayer-Pröschel M. Identifying the threshold of iron deficiency in the central nervous system of the rat by the auditory brainstem response. ASN Neuro. 2015; 7.

6. Tran PV, Fretham SJ, Wobken J, Miller BS, Georgieff MK. Gestationalneonatal iron deficiency suppresses and iron treatment reactivates IGF signaling in developing rat hippocampus. Am J Physiol Endocrinol Metab. 2012; 302:E316–E324.

7. Chung J, Wessling-Resnick M. Molecular mechanisms and regulation of iron transport. Crit Rev Clin Lab Sci. 2003; 40:151–182.

8. Gulec S, Anderson GJ, Collins JF. Mechanistic and regulatory aspects of intestinal iron absorption. Am J Physiol Gastrointest Liver Physiol. 2014; 307:G397–G409.

9. Sharp PA. Intestinal iron absorption: regulation by dietary & systemic factors. Int J Vitam Nutr Res. 2010; 80:231–242.

10. Shah YM, Matsubara T, Ito S, Yim SH, Gonzalez FJ. Intestinal hypoxiainducible transcription factors are essential for iron absorption following iron deficiency. Cell Metab. 2009; 9:152–164.

11. Srai SK, Chung B, Marks J, Pourvali K, Solanky N, Rapisarda C, Chaston TB, Hanif R, Unwin RJ, Debnam ES, Sharp PA. Erythropoietin regulates intestinal iron absorption in a rat model of chronic renal failure. Kidney Int. 2010; 78:660–667.

12. Hoki T, Miyanishi K, Tanaka S, Takada K, Kawano Y, Sakurada A, Sato M, Kubo T, Sato T, Sato Y, Takimoto R, Kobune M, Kato J. Increased duodenal iron absorption through up-regulation of divalent metal transporter 1 from enhancement of iron regulatory protein 1 activity in patients with nonalcoholic steatohepatitis. Hepatology. 2015; Forthcoming.

13. Galy B, Ferring-Appel D, Becker C, Gretz N, Gröne HJ, Schümann K, Hentze MW. Iron regulatory proteins control a mucosal block to intestinal iron absorption. Cell Rep. 2013; 3:844–857.

14. Bhaskaram P, Balakrishna N, Radhakrishna KV, Krishnaswamy K. Validation of hemoglobin estimation using Hemocue. Indian J Pediatr. 2003; 70:25–28.

15. Ahmed BS, Ahmed SN, Hassan HG. Ferritic as a potent marker of brest cancer. Int J Tech Res Appl. 2015; 3:184–187.

16. Torrance JD, Bothwell TH. A simple technique for measuring storage iron concentrations in formalinised liver samples. S Afr J Med Sci. 1968; 33:9–11.

17. Sim MO, Lee HI, Ham JR, Seo KI, Kim MJ, Lee MK. Anti-inflammatory and antioxidant effects of umbelliferone in chronic alcohol-fed rats. Nutr Res Pract. 2015; 9:364–369.

18. Min B, Lee H, Song JH, Han MJ, Chung J. Arctiin inhibits adipogenesis in 3T3-L1 cells and decreases adiposity and body weight in mice fed a high-fat diet. Nutr Res Pract. 2014; 8:655–661.

19. Lee SM, Lee SB, Prywes R, Vulpe CD. Iron deficiency upregulates Egr1 expression. Genes Nutr. 2015; 10:468.

20. Cairo G, Tacchini L, Pogliaghi G, Anzon E, Tomasi A, Bernelli-Zazzera A. Induction of ferritin synthesis by oxidative stress. Transcriptional and post-transcriptional regulation by expansion of the "free" iron pool. J Biol Chem. 1995; 270:700–703.

21. Dongiovanni P, Lanti C, Gatti S, Rametta R, Recalcati S, Maggioni M, Fracanzani AL, Riso P, Cairo G, Fargion S, Valenti L. High fat diet subverts hepatocellular iron uptake determining dysmetabolic iron overload. PLoS One. 2015; 10:e0116855.

22. Zhang AS, Enns CA. Molecular mechanisms of normal iron homeostasis. Hematology Am Soc Hematol Educ Program. 2009; 207–214.

23. Montalbetti N, Simonin A, Simonin C, Awale M, Reymond JL, Hediger MA. Discovery and characterization of a novel non-competitive inhibitor of the divalent metal transporter DMT1/SLC11A2. Biochem Pharmacol. 2015; 96:216–224.

24. Ikuta K, Yersin A, Ikai A, Aisen P, Kohgo Y. Characterization of the interaction between diferric transferrin and transferrin receptor 2 by functional assays and atomic force microscopy. J Mol Biol. 2010; 397:375–384.

25. Linder MC. Mobilization of stored iron in mammals: a review. Nutrients. 2013; 5:4022–4050.

26. Ganz T. Macrophages and systemic iron homeostasis. J Innate Immun. 2012; 4:446–453.

27. Nemeth E. Targeting the hepcidin-ferroportin axis in the diagnosis and treatment of anemias. Adv Hematol. 2010; 2010:750643.

28. De Domenico I, Ward DM, Langelier C, Vaughn MB, Nemeth E, Sundquist WI, Ganz T, Musci G, Kaplan J. The molecular mechanism of hepcidin-mediated ferroportin down-regulation. Mol Biol Cell. 2007; 18:2569–2578.

29. De Domenico I, Lo E, Ward DM, Kaplan J. Hepcidin-induced internalization of ferroportin requires binding and cooperative interaction with Jak2. Proc Natl Acad Sci U S A. 2009; 106:3800–3805.

30. Tsochatzis E, Papatheodoridis GV, Koliaraki V, Hadziyannis E, Kafiri G, Manesis EK, Mamalaki A, Archimandritis AJ. Serum hepcidin levels are related to the severity of liver histological lesions in chronic hepatitis C. J Viral Hepat. 2010; 17:800–806.

31. Xia Y, Babitt JL, Sidis Y, Chung RT, Lin HY. Hemojuvelin regulates hepcidin expression via a selective subset of BMP ligands and receptors independently of neogenin. Blood. 2008; 111:5195–5204.

32. Kautz L, Meynard D, Monnier A, Darnaud V, Bouvet R, Wang RH, Deng C, Vaulont S, Mosser J, Coppin H, Roth MP. Iron regulates phosphorylation of Smad1/5/8 and gene expression of Bmp6, Smad7, Id1, and Atoh8 in the mouse liver. Blood. 2008; 112:1503–1509.

33. Rausa M, Pagani A, Nai A, Campanella A, Gilberti ME, Apostoli P, Camaschella C, Silvestri L. Bmp6 expression in murine liver non parenchymal cells: a mechanism to control their high iron exporter activity and protect hepatocytes from iron overload? PLoS One. 2015; 10:e0122696.

34. Andriopoulos B Jr, Corradini E, Xia Y, Faasse SA, Chen S, Grgurevic L, Knutson MD, Pietrangelo A, Vukicevic S, Lin HY, Babitt JL. BMP6 is a key endogenous regulator of hepcidin expression and iron metabolism. Nat Genet. 2009; 41:482–487.

35. Fung E, Nemeth E. Manipulation of the hepcidin pathway for therapeutic purposes. Haematologica. 2013; 98:1667–1676.

36. Ramey G, Deschemin JC, Vaulont S. Cross-talk between the mitogen activated protein kinase and bone morphogenetic protein/hemojuvelin pathways is required for the induction of hepcidin by holotransferrin in primary mouse hepatocytes. Haematologica. 2009; 94:765–772.

37. Gao J, Chen J, De Domenico I, Koeller DM, Harding CO, Fleming RE, Koeberl DD, Enns CA. Hepatocyte-targeted HFE and TFR2 control hepcidin expression in mice. Blood. 2010; 115:3374–3381.