title: Immuno-spin trapping detection of antioxidant/pro-oxidant properties of zinc or selenium on DNA and protein radical formation via hydrogen peroxide.
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 Zn(2+) (100 µM) or SeO3(-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 Zn(2+) or SeO3(-2) are dual-functioning ions such as antioxidant at lower concentrations while pro-oxidant at higher concentrations. Particularly, our present data with SeO3(-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.
authors.affiliation: Department of Biophysics, Ankara University Faculty of Medicine, 06100, Ankara, Turkey.
authors.forename: Vedia
authors.initials: V
authors.lastname: Deletioglu
authors.affiliation: Department of Biophysics, Ankara University Faculty of Medicine, 06100, Ankara, Turkey.
authors.forename: Erkan
authors.initials: E
authors.lastname: Tuncay
authors.affiliation: Department of Biophysics, Ankara University Faculty of Medicine, 06100, Ankara, Turkey.
authors.forename: Aysegul
authors.initials: A
authors.lastname: Toy
authors.affiliation: Institute of Biomedicine, Physiology, University of Eastern Finland, Kuopio, Finland.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Atalay
authors.affiliation: Department of Biophysics, Ankara University Faculty of Medicine, 06100, Ankara, Turkey. belma.turan@medicine.ankara.edu.tr.
authors.forename: Belma
authors.initials: B
authors.lastname: Turan
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000737:chemistry*; D003497:Cyclic N-Oxides / Q000737:chemistry; D018736:DNA Adducts / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D006861:Hydrogen Peroxide / Q000737:chemistry*; D008297:Male; D024821:Metabolic Syndrome / Q000503:physiopathology; D023421:Models, Animal; D051381:Rats; D017208:Rats, Wistar; D012643:Selenium / Q000737:chemistry*; D013113:Spin Labels; D019097:Spin Trapping / Q000379:methods*; D015032:Zinc / Q000737:chemistry*
chemical_list: D000975:Antioxidants; D003497:Cyclic N-Oxides; D018736:DNA Adducts; D013113:Spin Labels; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D006861:Hydrogen Peroxide; D012643:Selenium; D015032:Zinc
keywords: Glutathione; Heart; Hydrogen peroxide; Lipid peroxidation; Metabolic syndrome; Reactive oxygen species