| Autor: |
Misak A; Biomedical Research Center, Department of Molecular Physiology, Institute of Clinical and Translational Research, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, Slovakia., Brezova V; Institute of Physical Chemistry and Chemical Physics, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, Radlinského 9, 81237 Bratislava, Slovakia., Chovanec M; Biomedical Research Center, Department of Genetics, Cancer Research Institute, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, Slovakia., Luspai K; Institute of Physical Chemistry and Chemical Physics, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, Radlinského 9, 81237 Bratislava, Slovakia., Nasim MJ; Division of Bioorganic Chemistry, School of Pharmacy, University of Saarland, D-66123 Saarbruecken, Germany., Grman M; Biomedical Research Center, Department of Molecular Physiology, Institute of Clinical and Translational Research, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, Slovakia., Tomasova L; Biomedical Research Center, Department of Molecular Physiology, Institute of Clinical and Translational Research, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, Slovakia., Jacob C; Division of Bioorganic Chemistry, School of Pharmacy, University of Saarland, D-66123 Saarbruecken, Germany., Ondrias K; Biomedical Research Center, Department of Molecular Physiology, Institute of Clinical and Translational Research, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, Slovakia. |
| Abstrakt: |
Superoxide radical anion (O 2 •- ) and its derivatives regulate numerous physiological and pathological processes, which are extensively studied. The aim of our work was to utilize KO 2 as a source of O 2 •- and the electron paramagnetic resonance (EPR) spin trapping 5- tert -butoxycarbonyl-5-methyl-1-pyrroline N -oxide (BMPO) technique for the preparation of • BMPO-OOH and/or • BMPO-OH radicals in water solution without DMSO. The method distinguishes the interactions of various compounds with • BMPO-OOH and/or • BMPO-OH radicals over time. Here, we show that the addition of a buffered BMPO-HCl mixture to powdered KO 2 formed relatively stable • BMPO-OOH and • BMPO-OH radicals and H 2 O 2 , where the • BMPO-OOH/OH ratio depended on the pH. At a final pH of ~6.5-8.0, the concentration of • BMPO-OOH radicals was ≥20 times higher than that of • BMPO-OH, whereas at pH 9.0-10.0, the • BMPO-OH radicals prevailed. The • BMPO-OOH/OH radicals effectively cleaved the plasmid DNA. H 2 S decreased the concentration of • BMPO-OOH/OH radicals, whereas the selenium derivatives 1-methyl-4-(3-(phenylselanyl) propyl) piperazine and 1-methyl-4-(4-(phenylselanyl) butyl) piperazine increased the proportion of • BMPO-OH over the • BMPO-OOH radicals. In conclusion, the presented approach of using KO 2 as a source of O 2 •- /H 2 O 2 and EPR spin trap BMPO for the preparation of • BMPO-OOH/OH radicals in a physiological solution could be useful to study the biological effects of radicals and their interactions with compounds. |
