Singlet oxygen-dependent hydroxyl radical formation during uroporphyrin-mediated photosensitization in the presence of NADPH.
The conversion of singlet oxygen ((1)O2) to hydroxyl radical (*OH) during photosensitization of uroporphyrin (UP) in the presence of NADPH was examined by a spin-trapping technique with 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). Significant electron spin resonance (ESR) signals of DMPO-OH adduct were observed during irradiation of the UP-NADPH system with visible light. Scavengers of *OH reduced the signal intensity to 3-30% of control, indicating that more than 70% of DMPO-OH results from freely diffusing *OH. The ESR signal was almost completely lost when quenchers of (1)O2 were added, and was enhanced when the amount of deutrated solvent was increased. The appearance of (1)O2, as determined by the oxidation of 2,2,6,6-tetramethyl-4-piperidone (TEMPD), was delayed with an increase in the concentration of NADPH, whereas the production of *OH was upregulated. These observations indicate that conversion of (1)O2 to *OH occurs quickly in the presence of NADPH. Hydrogen peroxide (H2O2) was produced (1)O2-dependently during irradiation of UP in the presence of NADPH. However, neither catalase nor desferrioxamine decreased the DMPO-OH signal, and addition of H2O2 did not increase the signal. SOD increased the signal only slightly. These results suggest that the production of *OH from (1)O2 involves neither superoxide anion radical nor H2O2.
Bioregulation Research Group, National Institute of Radiological Sciences, Chiba, Japan.
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Takeshita
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Olea-Azar
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Mizuno
T
T
Ozawa
D000818:Animals; D002374:Catalase / Q000378:metabolism / Q000494:pharmacology; D002417:Cattle; D003676:Deferoxamine / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D005730:Ganglionic Blockers / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D017665:Hydroxyl Radical / Q000378:metabolism*; D008027:Light; D008956:Models, Chemical; D009249:NADP / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D010881:Piperidones / Q000494:pharmacology; D013997:Time Factors; D014216:Triacetoneamine-N-Oxyl / Q000031:analogs & derivatives* / Q000494:pharmacology; D015854:Up-Regulation; D014578:Uroporphyrins / Q000378:metabolism*
D005730:Ganglionic Blockers; D010881:Piperidones; D014578:Uroporphyrins; D014216:Triacetoneamine-N-Oxyl; C002273:tempidon; D017665:Hydroxyl Radical; D009249:NADP; D006861:Hydrogen Peroxide; D002374:Catalase; D003676:Deferoxamine; D010100:Oxygen