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Effect of dietary protein on heme iron uptake by Caco-2 cells

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Abstract

Objective

To study heme iron bioavailability and the role of dietary protein (animal and vegetable) on iron uptake using an in vitro model (Caco-2 cell line).

Methods

Caco-2 cells were seeded in bicameral chambers with different animal (beef, chicken or fish) or vegetable (peas, lentils, and soybeans) proteins or with pure animal (collagen and casein) or vegetable (gliadin, zein, and glutein) protein extracts. The effect of each protein over heme iron absorption was assessed.

Results

Intact heme uptake was higher than either heme plus albumin or digested heme plus albumin, but lower than digested heme. White meal exerted the highest inhibitory effect on hemin uptake. Heme iron uptake decreased in the presence of all legume extracts, but was not significantly different among them (one-way ANOVA, NS). Pure animal (collagen and casein) and vegetable (zein and glutelin) proteins increased heme iron uptake, except for gliadin.

Conclusion

Animal and vegetable protein in general decreased heme iron uptake. However, purified animal and vegetable protein induce an increase in heme iron uptake.

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References

  1. McCord JM (1998). Iron, free radicals, and oxidative injury. Semin Hematol 35: 5–12. Review

    Google Scholar 

  2. Turnbull A, Cleton F, Finch CA (1962) Iron absorption. IV. The absorption of hemoglobin iron. J Clin Invest 41:1897–1907

    Article  CAS  Google Scholar 

  3. Hallberg L, Solvell L (1967) Absorption of hemoglobin iron in man. Acta Med Scand 181:335–354

    Article  CAS  Google Scholar 

  4. Callender ST, Mallett BJ, Smith MD (1957) Absorption of haemoglobin iron. Br J Haematol 3:186–192

    Article  CAS  Google Scholar 

  5. Conrad ME, Umbreit JN, Moore EG (1999) Iron absorption and transport. Am J Med Sci 318:213–229

    Article  CAS  Google Scholar 

  6. Conrad ME, Weintraub LR, Crosby WH (1964) The role of the intestine in iron kinetics. J Clin Invest 43:963–974

    Article  CAS  Google Scholar 

  7. Crosby WH, Conrad ME Jr, Wheby MS (1963) The rate of iron accumulation in iron storage disease. Blood 22:429–440

    CAS  Google Scholar 

  8. Carpenter CE, Mahoney AW (1992) Contributions of heme and nonheme iron to human nutrition. Crit Rev Food Sci Nutr 31:333–367

    Article  CAS  Google Scholar 

  9. Conrad ME, Benjamin BI, Williams HL, Foy AL (1967) Human absorption of hemoglobin-iron. Gastroenterology 53:5–10

    CAS  Google Scholar 

  10. Monsen ER, Hallberg L, Layrisse M, Hegsted DM, Cook JD, Mertz W, Finch CA (1978) Estimation of available dietary iron. Am J Clin Nutr 31:134–141

    CAS  Google Scholar 

  11. Worthington MT, Cohn SM, Miller SK, Qi Luo RQ, Berg CL (2001) Characterization of a human plasma membrane heme transporter in intestinal and hepatocyte cell lines. Am J Physiol Gastrointest Liver Physiol 280:G1172–G1177

    CAS  Google Scholar 

  12. Shayeghi M, Latunde-Dada GO, Oakhill JS, Laftah AH, Takeuchi K, Halliday N, Khan Y, Warley A, McCann FE, Hider RC, Frazer DM, Anderson GJ, Vulpe CD, Simpson RJ, McKie AT (2005) Identification of an intestinal heme transporter. Cell 122:789–801

    Article  CAS  Google Scholar 

  13. Bjorn-Rasmussen E, Hallberg L, Isaksson B, Arvidsson B (1974) Food iron absorption in man. Applications of the two-pool extrinsic tag method to measure heme and nonheme iron absorption from the whole diet. J Clin Invest 53:247–255

    Article  CAS  Google Scholar 

  14. Uzel C, Conrad ME (1998) Absorption of heme iron. Semin Hematol 35:27–34

    CAS  Google Scholar 

  15. Schricker BR, Miller DD (1983) Effects of cooking and chemical treatment on heme and non-heme iron in meat. J Food Sci 48:1340–1344

    Article  CAS  Google Scholar 

  16. Hallberg L, Rossander-Hulthen L, Brune M, Gleerup A (1993) Inhibition of haem-iron absorption in man by calcium. Br J Nutr 69:533–540

    Article  CAS  Google Scholar 

  17. Tenhunen R, Tokola O, Linden IB (1987) Haem arginate: a new stable haem compound. J Pharm Pharmacol 39:780–786

    Article  CAS  Google Scholar 

  18. Lee KS, Raymond LD, Schoen B, Raymond GJ, Kett L, Moore RA, Johnson LM, Taubner L, Speare JO, Onwubiko HA, Baron GS, Caughey WS, Caughey B (2007) Hemin interactions and alterations of the subcellular localization of prion protein. J Biol Chem 282:36525–36533

    Article  CAS  Google Scholar 

  19. Conrad ME, Umbreit JN (2000) Iron absorption and transport-an update. Am J Hematol 64:287–298

    Article  CAS  Google Scholar 

  20. Au AP, Reddy MB (2000) Caco-2 cells can be used to assess human iron bioavailability from a semipurified meal. J Nutr 130:1329–1334

    CAS  Google Scholar 

  21. Fogh J, Wright WC, Loveless JD (1977) Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J Natl Cancer Inst 58:209–214

    CAS  Google Scholar 

  22. Artursson P (1990) Epithelial transport of drugs in cell culture. I: a model for studying the passive diffusion of drugs over intestinal absorptive (Caco-2) cells. J Pharm Sci 79:476–482

    Article  CAS  Google Scholar 

  23. Alvarez-Hernandez X, Nichols GM, Glass J (1991) Caco-2 cell line: a system for studying intestinal iron transport across epithelial cell monolayers. J Biochim Biophys Acta 1070:205–208

    Article  CAS  Google Scholar 

  24. Han O, Failla M, Hill A, Morris E, Smith J (1995) Reduction of Fe(III) is required for uptake of nonheme iron by Caco-2 cells. J Nutr 125:1291–1299

    CAS  Google Scholar 

  25. Nuñez MT, Alvarez X, Smith M, Tapia V, Glass J (1994) Role of redox systems on Fe3 + uptake by transformed human intestinal epithelial (Caco-2) cells. Am J Physiol 267:C1582–C1588

    Google Scholar 

  26. Alvarez-Hernandez X, Smith M, Glass J (1994) Regulation of iron uptake and transport by transferrin in Caco-2 cells, an intestinal cell line. Biochim Biophys Acta 1192:215–222

    Article  CAS  Google Scholar 

  27. Gangloff M, Lai C, Van Campen DR, Miller DD, Glahn RP (1996) Ferrous iron uptake but not transfer is down-regulated in Caco-2 cells grown in high iron serum-free medium. J Nutr 126:3118–3127

    CAS  Google Scholar 

  28. Nunez MT, Tapia V (1999) Transferrin stimulates iron absorption, exocytosis, and secretion in cultured intestinal cells. Am J Physiol 276:C1085–C1090

    CAS  Google Scholar 

  29. Garcia MN, Flowers C, Cook JD (1996) The Caco-2 cell culture system can be used as a model to study food iron availability. J Nutr 126:251–258

    CAS  Google Scholar 

  30. Glahn RP, Lai C, Hsu J, Thompson JF, Guo M, Van Campen DR (1998) Decreased citrate improves iron availability from infant formula: application of an in vitro digestion/Caco-2 cell culture model. J Nutr 128:257–264

    CAS  Google Scholar 

  31. Glahn RP, Lee OA, Yeung A, Goldman MI, Miller DD (1998) Caco-2 cell ferritin formation predicts nonradiolabeled food iron availability in an in vitro digestion/Caco-2 cell culture model. J Nutr 128:1555–1561

    CAS  Google Scholar 

  32. Arredondo M, Kloosterman J, Núñez S, Segovia F, Candia V, Flores S, Le Blanc S, Olivares M, Pizarro F (2008) Heme iron uptake by Caco-2 cells is a saturable, temperature sensitive and modulated by extracellular pH and potassium. Biol Trace Elem Res 125:109–119

    Article  CAS  Google Scholar 

  33. Chaney AL, Marbach EP (1962) Modified reagents for determination of urea and ammonia. Clin Chem 8:130–132

    CAS  Google Scholar 

  34. Olivares M, Pizarro F, de Pablo S, Araya M, Uauy R (2004) Iron, zinc, and copper: contents in common Chilean foods and daily intakes in Santiago, Chile. Nutrition 20:205–212

    Article  CAS  Google Scholar 

  35. Rebouche CJ, Wilcox CL, Widness JA (2004) Microanalysis of non-heme iron in animal tissues. J Biochem Biophys Methods 58:239–251

    Article  CAS  Google Scholar 

  36. Udaka J, Koga T, Tsuji H, Kimoto M, Takumi K (2000) Efficient extraction and some properties of storage proteins (prolamin and glutelin) in ancient rice cultivars. J Nutr Sci Vitaminol (Tokyo) 46:84–90

    Article  CAS  Google Scholar 

  37. Bamdad F, Hossein Goli A, Kadivar M (2006) Preparation and characterization of proteinous film from lentil (Lens culinaris): Edible film from lentil (Lens culinaris) Food. Res. Internat 39:106–111

    CAS  Google Scholar 

  38. Tomoskozi S, Lasztity R, Haraszi R, Baticz O (2001) Isolation and study of the functional properties of pea proteins. Nahrung 45:399–401

    Article  CAS  Google Scholar 

  39. Lombardo ME, Lombardo ME, Araujo LS, Ciccarelli AB, Battle A (2005) A spectrophotometric method for estimating hemin in biological systems. Anal Biochem 341:199–203

    Article  CAS  Google Scholar 

  40. Lowry OH, Rosebrough N, Farr A, Randall R (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  Google Scholar 

  41. Layrisse M, Martinez-Torres C, Roche M (1968) Effect of interaction of various foods on iron absorption. Am J Clin Nutr 21:1175–1183

    CAS  Google Scholar 

  42. Layrisse M, Cook JD, Martinez-Torres C, Roche M, Kuhn IN, Finch CA (1969) Food iron absorption: a comparison of vegetable and animal foods. Blood 33:430–443

    CAS  Google Scholar 

  43. Hallberg L, Bjom-Rasmussen E, Howard L, Rossander L (1979) Dietary heme iron absorption. A discussion of possible mechanisms for the absorption-promoting effect of meat and for the regulation of iron absorption. Scand J Gastroenterol 14:769–779

    Article  CAS  Google Scholar 

  44. Bjorn-Rasmussen E, Hallberg L (1979) Effect of animal proteins on the absorption of food iron in man. Nutr Metab 23:192–202

    Article  CAS  Google Scholar 

  45. Lynch SR, Dassenko SA, Morck TA, Beard JL, Cook JD (1985) Soy protein products and heme iron absorption in humans. Am J Clin Nutr 41:13–20

    CAS  Google Scholar 

  46. Vaghefi N, Nedjaoum F, Guillochon D, Bureau F, Arhan P, Bouglé D (2002) Influence of the extent of hemoglobin hydrolysis on the digestive absorption of heme iron. An in vitro study. J Agric Food Chem 50:4969–4973

    Article  CAS  Google Scholar 

  47. Dang TN, Bishop GM, Dringen R, Robinson SR (2010) The putative heme transporter HCP1 is expressed in cultured astrocytes and contributes to the uptake of hemin. Glia 58:55–65

    Article  Google Scholar 

  48. Layrisse M, Martinez-Torres C (1972) Model for measuring dietary absorption of heme iron: test with a complete meal. Am J Clin Nutr 25:401–411

    CAS  Google Scholar 

  49. Conrad ME, Weintraub LR, Sears DA, Crosby WH (1966) Absorption of hemoglobin iron. Am J Physiol 211:1123–1130

    CAS  Google Scholar 

  50. Hazell T, Ledward DA, Neale RJ (1978) Iron availability from meat. Br J Nutr 39:631–638

    Article  CAS  Google Scholar 

  51. Hazell T, Ledward DA, Neale RJ (1982) Hemoglobin and meat iron absorption. Am J Clin Nutr 36:187–189

    CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful for support from Fondo Nacional de Desarrollo Científico y Tecnológico de Chile (FONDECYT), grant number 1095038.

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Authors and Affiliations

  1. Micronutrient Laboratory, INTA, University of Chile, El Líbano 5524, 138-11, Macul, Santiago, Chile

    Pía Villarroel, Sebastián Flores, Fernando Pizarro, Daniel López de Romaña & Miguel Arredondo

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  1. Pía Villarroel
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  2. Sebastián Flores
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  3. Fernando Pizarro
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  5. Miguel Arredondo
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Correspondence to Miguel Arredondo.

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Villarroel, P., Flores, S., Pizarro, F. et al. Effect of dietary protein on heme iron uptake by Caco-2 cells. Eur J Nutr 50, 637–643 (2011). https://doi.org/10.1007/s00394-011-0173-8

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  • DOI: https://doi.org/10.1007/s00394-011-0173-8

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