title: Antioxidant paradoxes of phenolic compounds: peroxyl radical scavenger and lipid antioxidant, etoposide (VP-16), inhibits sarcoplasmic reticulum Ca(2+)-ATPase via thiol oxidation by its phenoxyl radical.
abstract: The effectiveness of a phenolic antioxidant as a radical scavenger is determined by its reactivity toward peroxyl radicals and also by the reactivity of the anti-oxidant phenoxyl radical toward oxidation substrate. If the phenoxyl radical efficiently interacts with vitally important biomolecules, this interaction may result in oxidative damage rather than antioxidant protection. In the present work, we studied effects of phenoxyl radicals generated from a phenolic antitumor drug, Etoposide (VP-16), on oxidation of thiols and activity of Ca(2+)-ATPase in sarcoplasmic reticulum (SR) membranes from skeletal muscles. We found that VP-16 is an effective scavenger of peroxyl radicals as judged by its ability to inhibit a water-soluble azo-initiator, 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH)-induced (i) chemiluminescence (oxidation) of luminol, (ii) fluorescence decay (oxidation) of cis-parinaric acid incorporated in SR membranes, and (iii) peroxidation of SR membrane lipids. VP-16 did not prevent AAPH-induced oxidation of sulfhydryl groups and inhibition of Ca(2+)-ATPase in SR membranes. Electron spin resonance measurements showed that AAPH-induced VP-16 phenoxyl radicals were reduced by interaction with SR thiols. By using tyrosinase to generate VP-16 phenoxyl radicals as the only source of free radicals in the model system, we found that inhibition of Ca(2+)-ATPase was accompanied by oxidation of about 5 mol of Ca(2+)-ATPase SH groups per 1 mol of oxidized VP-16. Secondary products of VP-16 oxidation (including VP-16 o-quinone) were not efficient in inhibiting SR Ca(2+)-ATPase. Reduction of VP-16 phenoxyl radicals by ascorbate protected against AAPH- and tyrosinase-induced thiol oxidation and Ca(2+)-ATPase inhibition. The results suggest that efficient phenolic scavengers of peroxyl radicals such as VP-16--which are commonly considered as potent antioxidants--may themselves produce oxidative stress due to secondary reactions of their phenoxyl radicals with thiols.
authors.affiliation: Department of Environmental and Occupational Health, University of Pittsburgh, Pennsylvania 15238, USA.
authors.forename: V B
authors.initials: VB
authors.lastname: Ritov
authors.forename: R
authors.initials: R
authors.lastname: Goldman
authors.forename: D A
authors.initials: DA
authors.lastname: Stoyanovsky
authors.forename: E V
authors.initials: EV
authors.lastname: Menshikova
authors.forename: V E
authors.initials: VE
authors.lastname: Kagan
mesh_terms: D000408:Alamethicin / Q000494:pharmacology; D000578:Amidines; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D002374:Catalase / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D005047:Etoposide / Q000494:pharmacology*; D005231:Fatty Acids, Unsaturated; D016166:Free Radical Scavengers / Q000494:pharmacology*; D005609:Free Radicals / Q000494:pharmacology; D007700:Kinetics; D008163:Luminescent Measurements; D018482:Muscle, Skeletal / Q000201:enzymology; D010084:Oxidation-Reduction; D010636:Phenols / Q000494:pharmacology*; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D013050:Spectrometry, Fluorescence; D013482:Superoxide Dismutase / Q000494:pharmacology; D013997:Time Factors
chemical_list: D000578:Amidines; D000975:Antioxidants; D005231:Fatty Acids, Unsaturated; D016166:Free Radical Scavengers; D005609:Free Radicals; D010636:Phenols; D000408:Alamethicin; C042329:phenoxy radical; D005047:Etoposide; C046728:2,2'-azobis(2-amidinopropane); D002374:Catalase; D013482:Superoxide Dismutase; D000252:Calcium-Transporting ATPases; C013641:parinaric acid