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Iron toxicity mediated by oxidative stress enhances tissue damage in an animal model of diabetes

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Abstract

Although iron is a first-line pro-oxidant that modulates clinical manifestations of various systemic diseases, including diabetes, the individual tissue damage generated by active oxidant insults has not been demonstrated in current animal models of diabetes. We tested the hypothesis that oxidative stress is involved in the severity of the tissues injury when iron supplementation is administered in a model of type 1 diabetes. Streptozotocin (Stz)-induced diabetic and non-diabetic Fischer rats were maintained with or without a treatment consisting of iron dextran ip at 0.1 mL day−1 doses administered for 4 days at intervals of 5 days. After 3 weeks, an extensive increase (p < 0.001) in the production of reactive oxygen species (ROS) in neutrophils of the diabetic animals on iron overload was observed. Histological analysis revealed that this treatment also resulted in higher (p < 0.05) tissue iron deposits, a higher (p < 0.001) number of inflammatory cells in the pancreas, and apparent cardiac fibrosis, as shown by an increase (p < 0.05) in type III collagen levels, which result in dysfunctional myocardial. Carbonyl protein modification, a marker of oxidative stress, was consistently higher (p < 0.01) in the tissues of the iron-treated rats with diabetes. Moreover, a significant positive correlation was found between ROS production and iron pancreas stores (r = 0.42, p < 0.04), iron heart stores (r = 0.54, p < 0.04), and change of the carbonyl protein content in pancreas (r = 0.49, p < 0.009), and heart (r = 0.48, p < 0.02). A negative correlation was still found between ROS production and total glutathione content in pancreas (r = −0.50, p < 0.03) and heart (r = −0.45, p < 0.04). In conclusion, our results suggest that amplified toxicity in pancreatic and cardiac tissues in rats with diabetes on iron overload might be attributed to increased oxidative stress.

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Abbreviations

AGEs:

Advanced glycation end products

ANOVA:

Analysis of variance

AOAC:

Association of official analytical chemists

BSA:

Bovine serum albumin

C:

Control group

CAT:

Catalase

CI:

Control iron group

DM:

Diabetes mellitus

D:

Diabetic group

DI:

Diabetic iron group

DNPH:

2,4-Dinitrophenylhydrazine

DPI:

Diphenylene iodonium

DTNB:

5,5-Dithiobis(2-nitrobenzoic acid)

GSH-Px:

Glutathione peroxidase

GSSG:

Oxidized glutathione

GSH:

Reduced glutathione

H2O2 :

Hydrogen peroxide

LIBC:

Latent iron-binding capacity

OH− :

Hydroxyl radicals

O2 − :

Superoxide anions

PB:

Pearl’s Prussian blue

PPAR-α:

Peroxisome proliferator activated receptor-α

RLU/min:

Relative units of light/min

ROS:

Reactive oxygen species

Stz:

Streptozotocin

SOD:

Superoxide dismutase

TCA:

Trichloroacetic acid

TIBC:

Total iron-binding capacity

TNB:

5-Thio-2-nitrobenzoic acid

ZC3b:

Zymosan

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Acknowledgments

This work received financial support from Fundação de Amparo à Pesquisa do Estado de Minas Gerais, Universidade Federal de Ouro Preto and research fellowships from Conselho Nacional de Desenvolvimento Científico e Tecnológico (Pedrosa, M. L., Silva, M. E.) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Sampaio, A. F. S., Silva, M., Dornas, W.C.).

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Author notes

  1. Waleska C. Dornas

    Present address: Research Center in Biological Sciences (NUPEB), Universidade Federal de Ouro Preto, Campus Morro do Cruzeiro s/n, Ouro Preto, Minas Gerais, 35400-000, Brazil

Authors and Affiliations

  1. Program of Health and Nutrition, Federal University of Ouro Preto, Ouro Preto, Minas Gerais, Brazil

    Ana Flávia S. Sampaio

  2. Research Center in Biological Sciences (NUPEB), Universidade Federal de Ouro Preto, Campus Morro do Cruzeiro s/n, Ouro Preto, Minas Gerais, 35400-000, Brazil

    Maisa Silva, Daniela C. Costa, Marcelo E. Silva, Wanderson G. de Lima & Maria Lúcia Pedrosa

  3. Department of Foods, Federal University of Ouro Preto, Ouro Preto, Minas Gerais, Brazil

    Marcelo E. Silva & Rinaldo C. dos Santos

  4. Department of Biological Sciences, Federal University of Ouro Preto, Ouro Preto, Minas Gerais, Brazil

    Daniela C. Costa, Wanderson G. de Lima & Maria Lúcia Pedrosa

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  1. Ana Flávia S. Sampaio
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  2. Maisa Silva
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  8. Maria Lúcia Pedrosa
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Correspondence to Maria Lúcia Pedrosa.

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Sampaio, A.F.S., Silva, M., Dornas, W.C. et al. Iron toxicity mediated by oxidative stress enhances tissue damage in an animal model of diabetes. Biometals 27, 349–361 (2014). https://doi.org/10.1007/s10534-014-9717-8

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  • DOI: https://doi.org/10.1007/s10534-014-9717-8

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