Skip to main content
Log in

Oxidative Stress and Genomic Damage Induced In Vitro in Human Peripheral Blood by Two Preventive Treatments of Iron Deficiency Anemia

  • Published:
Biological Trace Element Research Aims and scope Submit manuscript

Abstract

Iron deficiency is the most prevalent nutritional deficiency and the main cause of anemia worldwide. Since children aged 6–24 months are among the most vulnerable groups at risk, daily supplementation with ferrous sulfate is recommended by the Argentine Society of Pediatrics as preventive treatment of anemia. However, a single weekly dose would have fewer adverse side effects and has been therefore proposed as an alternative treatment. Ferrous sulfate is known by its pro-oxidative properties, which may lead to increased oxidative stress as well as lipid, protein, and DNA damage. We analyzed the effect of daily and weekly preventive treatment of iron deficiency anemia (IDA) on cell viability, oxidative stress, chromosome, and cytomolecular damage in peripheral blood cultured in vitro. The study protocol included the following: untreated negative control; bleomycin, hydrogen peroxide, or ethanol-treated positive control; daily 0.14 mg ferrous sulfate–supplemented group; and weekly 0.55 mg ferrous sulfate–supplemented group. We assessed cell viability (methyl-thiazolyl-tetrazolium and neutral red assays), lipid peroxidation (thiobarbituric acid reactive substances assay), antioxidant response (superoxide dismutase and catalase enzyme analysis), chromosome damage (cytokinesis-blocked micronucleus cytome assay), and cytomolecular damage (comet assay). Lipid peroxidation, antioxidant response, and chromosome and cytomolecular damage decreased after weekly ferrous sulfate supplementation (p < 0.05), suggesting less oxygen free radical production and decreased oxidative stress and genomic damage. Such a decrease in oxidative stress and genomic damage in vitro positions weekly supplementation as a better alternative for IDA treatment. Further studies in vivo would be necessary to corroborate whether weekly supplementation could improve IDA preventive treatment compliance in children.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Watkins ML, Erickson JD, Thun MJ, Mulinare J, Heath CW (2000) Multivitamin use and mortality in a large prospective study. Am J Epidemiol 152:149–162

    Article  CAS  PubMed  Google Scholar 

  2. Fenech M (2001) Recommended dietary allowances (RDAs) for genomic stability. Mutat Res 480–481:51–54

    Article  PubMed  Google Scholar 

  3. Fenech M (2005) The Genome Health Clinic and Genome Health Nutrigenomics concepts: diagnosis and nutritional treatment of genome and epigenome damage on an individual basis. Mutagenesis 20:255–269. https://doi.org/10.1093/mutage/gei040

    Article  CAS  PubMed  Google Scholar 

  4. Fenech MF (2014) Nutriomes and personalised nutrition for DNA damage prevention, telomere integrity maintenance and cancer growth control. In: Zappia V, Panico S, Russo GL, Budillon A, Della Ragione F (eds) Advances in nutrition and cancer. Springer Berlin Heidelberg, Berlin, pp 427–441. https://doi.org/10.1007/978-3-642-38007-5_24 (accessed November 6, 2017)

    Chapter  Google Scholar 

  5. Beard JL (2001) Iron biology in immune function, muscle metabolism and neuronal functioning. J Nutr 131:568S–579S discussion 580S

    Article  CAS  PubMed  Google Scholar 

  6. Lönnerdal B (2017) Excess iron intake as a factor in growth, infections, and development of infants and young children. Am J Clin Nutr 106:1681S–1687S. https://doi.org/10.3945/ajcn.117.156042

    Article  PubMed  PubMed Central  Google Scholar 

  7. Aksu BY, Hasbal C, Himmetoglu S, Dincer Y, Koc EE, Hatipoglu S, Akcay T (2010) Leukocyte DNA damage in children with iron deficiency anemia: effect of iron supplementation. Eur J Pediatr 169:951–956. https://doi.org/10.1007/s00431-010-1147-1

    Article  CAS  PubMed  Google Scholar 

  8. Camaschella C (2017) New insights into iron deficiency and iron deficiency anemia. Blood Rev 31:225–233. https://doi.org/10.1016/j.blre.2017.02.004

    Article  CAS  PubMed  Google Scholar 

  9. González HF, Malpeli A, Etchegoyen G, Lucero L, Romero F, Lagunas C, Lailhacar G, Olivares M, Uauy R (2007) Acquisition of visuomotor abilities and intellectual quotient in children aged 4–10 years: relationship with micronutrient nutritional status. Biol Trace Elem Res 120:92–101. https://doi.org/10.1007/s12011-007-8023-5

    Article  CAS  PubMed  Google Scholar 

  10. Clark SF (2008) Iron deficiency anemia. Nutr Clin Pract 23:128–141. https://doi.org/10.1177/0884533608314536

    Article  PubMed  Google Scholar 

  11. International Nutritional Anemia Consultative Group: PAHO, UNICEF, WB, MI, USAID, FAO, Anemia prevention and control: what works-part 1 and part 2 (2003)

  12. Encuesta Nacional de Nutrición y Salud. Documento de Resultados. Buenos Aires (2007). Ministerio de Salud de la Nación. http://www.msal.gov.ar (accessed 1 May 2017)

  13. Apezteguia MC, Varea A, Disalvo L, Malpeli A, González HF (2008) Deficiencia de micronutrientes en niños de 1 a 3 años de familias de bajos ingresos en dos regiones de la provincia de Buenos Aires (Argentina). XLVI Reunión Anual de la Sociedad Latinoamericana de Investigación Pediátrica 38

  14. Ianicelli J (2012) Prevalencia de anemia en lactantes menores de 6 meses asistidos en un centro de atención primaria de la ciudad de La Plata. Archivos Argentinos de Pediatria 110:120–125. https://doi.org/10.5546/aap.2012.120

    Article  PubMed  Google Scholar 

  15. UNICEF/UNU/WHO (2001) Iron deficiency anaemia. Assessment, prevention and control. A guide for programme managers. WHO/NHD/01.3. Geneva: WHO

  16. Sociedad Argentina de Pediatría. Archivos Argentinos de Pediatría (2011) Anemia ferropénica. Normas de diagnóstico y tratamiento 99(2):62–166

    Google Scholar 

  17. Zalles Cueto L, Rojas Meneces J, Rojas Soto S, Sejas E (2005) Eficacia de la suplementación semanal versus diaria con sulfato ferroso en niños escolares con anemia ferropenica. Gac Med Bol 28(2):3–8

    Google Scholar 

  18. Tee ES, Kandiah M, Awin N, Chong SM, Satgunasingam N, Kamarudin L, Milani S, Dugdale AE, Viteri FE (1999) School-administered weekly iron-folate supplements improve hemoglobin and ferritin concentrations in Malaysian adolescent girls. Am J Clin Nutr 69:1249–1256

    Article  CAS  PubMed  Google Scholar 

  19. Agarwal KN, Gomber S, Bisht H, Som M (2003) Anemia prophylaxis in adolescent school girls by weekly or daily iron-folate supplementation. Indian Pediatr 40:296–301

    CAS  PubMed  Google Scholar 

  20. Viteri FE, Ali F, Tujague J (1999) Long-term weekly iron supplementation improves and sustains nonpregnant women’s iron status as well or better than currently recommended short-term daily supplementation. J Nutr 129:2013–2020

    Article  CAS  PubMed  Google Scholar 

  21. Peña Rosas JP, De Regil LM, Gomez Malave H, Flores Urrutia MC, Dowswell T (2015) Intermittent oral iron supplementation during pregnancy. In: The Cochrane Collaboration (ed), Cochrane Database of Systematic Reviews, John Wiley & Sons, Ltd, Chichester, UK. https://doi.org/10.1002/14651858.CD009997.pub2 (accessed May 18, 2018)

  22. Andersen HS, Gambling L, Holtrop G, McArdle HJ (2006) Maternal iron deficiency identifies critical windows for growth and cardiovascular development in the rat postimplantation embryo. J Nutr 136(5):1171–1177

    Article  CAS  PubMed  Google Scholar 

  23. Viteri FE (1997) Iron supplementation for the control of iron deficiency in populations at risk. Nutr Rev 55(6):195–209

    Article  CAS  PubMed  Google Scholar 

  24. Prá D, Franke SIR, Giulian R, Yoneama ML, Dias JF, Erdtmann B, Henriques JAP (2008) Genotoxicity and mutagenicity of iron and copper in mice. Biometals 21:289–297. https://doi.org/10.1007/s10534-007-9118-3

    Article  CAS  PubMed  Google Scholar 

  25. Puntarulo S (2005) Iron, oxidative stress and human health. Mol Asp Med 26:299–312. https://doi.org/10.1016/j.mam.2005.07.001

    Article  CAS  Google Scholar 

  26. Franke SIR, Prá D, da Silva J, Erdtmann B, Henriques JAP (2005) Possible repair action of vitamin C on DNA damage induced by methyl methanesulfonate, cyclophosphamide, FeSO4 and CuSO4 in mouse blood cells in vivo. Mutat Res Genet Toxicol Environ Mutagen 583:75–84. https://doi.org/10.1016/j.mrgentox.2005.03.001

    Article  CAS  Google Scholar 

  27. Prá D, Franke SIR, Henriques JAP, Fenech M (2012) Iron and genome stability: an update. Mutat Res- Fund Mol M 733:92–99. https://doi.org/10.1016/j.mrfmmm.2012.02.001

    Article  CAS  Google Scholar 

  28. Aslan M, Horoz M, Kocyigit A, Ozgonül S, Celik H, Celik M, Erel O (2006) Lymphocyte DNA damage and oxidative stress in patients with iron deficiency anemia. Mutat Res-Fund Mol M 601:144–149. https://doi.org/10.1016/j.mrfmmm.2006.06.013

    Article  CAS  Google Scholar 

  29. Fenech MF (2010) Nutriomes and nutrient arrays - the key to personalised nutrition for DNA damage prevention and cancer growth control. Genome Integr 1:11. https://doi.org/10.1186/2041-9414-1-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Berger J, Aguayo VM, Tellez W, Lujan C, Traissac P, San Miguel JL (1997) Weekly iron supplementation is as effective as 5 day per week iron supplementation in Bolivian school children living at high altitude. Eur J Clin Nutr 51(6):381–386

    Article  CAS  PubMed  Google Scholar 

  31. Borenfreund E, Puerner JA (1985) Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicol Lett 24:119–124

    Article  CAS  PubMed  Google Scholar 

  32. Fenech M (2007) Cytokinesis-block micronucleus cytome assay. Nat Protoc 2:1084–1104. https://doi.org/10.1038/nprot.2007.77

    Article  CAS  PubMed  Google Scholar 

  33. Singh NP, McCo MT, Tice RR, Schneider EL (1988) A simple technique for quantification of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191

    Article  CAS  PubMed  Google Scholar 

  34. Olive PL (1999) DNA damage and repair in individual cells: applications of the comet assay in radiobiology. Int J Radiat Biol 75(4):395–405

    Article  CAS  PubMed  Google Scholar 

  35. Collins AR (2004) The comet assay for DNA damage and repair. Mol Biotechnol 26(3):249–261

    Article  CAS  PubMed  Google Scholar 

  36. Jover R, Ponsoda X, Castell JV, Gómez-Lechón MJ (1992) Evaluation of the cytotoxicity of ten chemicals on human cultured hepatocytes: predictability of human toxicity and comparison with rodent cell culture systems. Toxicol in Vitro 6:47–52. https://doi.org/10.1016/0887-2333(92)90084-5

    Article  CAS  PubMed  Google Scholar 

  37. Alcântara DDFA, Ribeiro HF, Matos LA, Sousa JMC, Burbano RR, Bahia MO (2013) Cellular responses induced in vitro by iron (Fe) in a central nervous system cell line (U343MGa). Genet Mol Res 12:1554–1560. https://doi.org/10.4238/2013.May.13.9

    Article  CAS  PubMed  Google Scholar 

  38. Hacıhamdioglu DÖ, Kurekci AE, Gursel O, Atay AA, Balamtekin N, Aydın A, Haşimi A, Ozcan O (2013) Evaluation of lipid peroxidation and antioxidant system in healthy iron-replete infants receiving iron prophylaxis. Nutrition 29:138–142. https://doi.org/10.1016/j.nut.2012.05.009

    Article  CAS  PubMed  Google Scholar 

  39. Zaka-Ur-Rab Z, Adnan M, Ahmad SM, Islam N (2016) Effect of oral iron on markers of oxidative stress and antioxidant status in children with iron deficiency anaemia. J Clin Diagn Res 10:(10)SC13-SC13, SC19.

  40. Kurtoglu E, Ugur A, Baltaci AK, Undar L (2003) Effect of iron supplementation on oxidative stress and antioxidant status in iron-deficiency anemia. Biol Trace Elem Res 96(1–3):117–123

    Article  CAS  PubMed  Google Scholar 

  41. Altun D, Kurekci AE, Gursel O, Hacıhamdioglu DO, Kurt I, Aydın A, Ozcan O (2014) Malondialdehyde, antioxidant enzymes, and renal tubular functions in children with iron deficiency or iron-deficiency anemia. Biol Trace Elem Res 161(1):48–56

    Article  CAS  PubMed  Google Scholar 

  42. National Academy of Sciences (2004) Food and Nutrition Board, Institute of Medicine. Dietary Reference Intakes (DRIs) tolerable upper intake levels, elements −1997-2001. (accessed May, 20 2018)

  43. Premkumar K, Bowlus CL (2003) Ascorbic acid reduces the frequency of iron induced micronuclei in bone marrow cells of mice. Mutat Res-Genet Toxicol Environ Mutagen 542(1):99–103. https://doi.org/10.1016/j.mrgentox.2003.09.002

    Article  CAS  Google Scholar 

  44. Ferro E, Visalli G, La Rosa MA, Piraino B, Civa R, Randazzo Papa G, Di Pietro A (2016) Genotoxic effect of iron overload and disease complications in transfused β thalassaemic patients. Mutagenesis 32:275–281. https://doi.org/10.1093/mutage/gew062.

    Article  Google Scholar 

  45. Mollet IG, Patel D, Govani FS, Giess A, Paschalaki K, Periyasamy M, Lidington EC, Mason JC, Jones MD, Game L, Ali S, Shovlin CL (2016) Low dose iron treatments induce a DNA damage response in human endothelial cells within minutes. PLoS One 11:e0147990. https://doi.org/10.1371/journal.pone.0147990

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors thank Adriana Di Maggio for careful manuscript translation and editing and César E. Bianchi for technical assistance.

Funding

This study was supported with funds provided by the National Scientific and Technical Research Council of Argentina (Grant No. 0657) and La Plata National University (Grants V246 and V249).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Rocío Celeste Gambaro.

Ethics declarations

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Declarations of Interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gambaro, R.C., Seoane, A. & Padula, G. Oxidative Stress and Genomic Damage Induced In Vitro in Human Peripheral Blood by Two Preventive Treatments of Iron Deficiency Anemia. Biol Trace Elem Res 190, 318–326 (2019). https://doi.org/10.1007/s12011-018-1576-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12011-018-1576-7

Keywords

Navigation