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Immuno-spin trapping detection of antioxidant/pro-oxidant properties of zinc or selenium on DNA and protein radical formation via hydrogen peroxide

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Abstract

Trace elements can participate in the catalysis of group-transfer reactions and can serve as their structural components. However, most of them including zinc and selenium have multifunctional roles in biological environments such as antioxidant and/or pro-oxidant effects, as concentration-dependent manner. Although it has been demonstrated the antioxidant actions of either selenium or zinc compounds, there are several documents pointing out their pro-oxidant/oxidant roles in biological systems. Here we have used ELISA-based immuno-spin trapping, a method for detection of free radical formation, to detect whether or not a zinc compound, Zn3(PO4)2, or a selenium compound, Na2SeO3, has antioxidant and/or pro-oxidant effect on 5,5-Dimethyl-1-Pyrroline-N-Oxide (DMPO)-DNA nitrone adducts induced with Cu(II)-H2O2-oxidizing system in in vitro preparations. Second, we examined whether this technique is capable to demonstrate the different DMPO-protein nitrone adduct productions in isolated protein crude of hearts from normal rats (CON) or rats with metabolic syndrome (MetS). Our data demonstrated that either Zn2+ (100 µM) or \({\text{SeO}}_{3}^{ - 2}\) (50 nM) has very strong antioxidant action against 200 µM H2O2-induced DMPO-DNA nitrone adduct production, whereas their higher concentrations have apparent pro-oxidant actions. We also used verification by Western blotting analysis whether immuno-spin trapping can be used to assess H2O2-induced DMPO-protein nitrone adducts in heart protein crudes. Our Western blot data further confirmed the ELISA-data from proteins and demonstrated how Zn2+ or \({\text{SeO}}_{3}^{ - 2}\) are dual-functioning ions such as antioxidant at lower concentrations while pro-oxidant at higher concentrations. Particularly, our present data with \({\text{SeO}}_{3}^{ - 2}\) in DMPO-protein nitrone adducts, being in line with our previous observation on its dual-actions in ischemia/reperfusion-induced damaged heart, have shown that this ion has higher pro-oxidant actions over 50 nM in MetS-group compared to that of CON group.

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Acknowledgments

This work was supported by grant of TUBITAK (SBAG-113S466) to Belma Turan as well as a collaborative study of Belma Turan and Mustafa Atalay in COST Action TD1304. Vedia Deletioglu was supported with ERASMUS programme between Turkey and Finland. The authors would like to thank Drs. R.P. Mason, M. Ehrenshaft, and F.A. Summers for their very kind technical supports.

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Authors and Affiliations

  1. Department of Biophysics, Ankara University Faculty of Medicine, 06100, Ankara, Turkey

    Vedia Deletioglu, Erkan Tuncay, Aysegul Toy & Belma Turan

  2. Institute of Biomedicine, Physiology, University of Eastern Finland, Kuopio, Finland

    Mustafa Atalay

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  1. Vedia Deletioglu
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Deletioglu, V., Tuncay, E., Toy, A. et al. Immuno-spin trapping detection of antioxidant/pro-oxidant properties of zinc or selenium on DNA and protein radical formation via hydrogen peroxide. Mol Cell Biochem 409, 23–31 (2015). https://doi.org/10.1007/s11010-015-2508-x

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