Antioxidant effect of ethanol toward in vitro peroxidation of human low-density lipoproteins initiated by oxygen free radicals.

This study was designed to evaluate the effect of ethanol on the peroxidation of human low-density lipoprotein (LDL) initiated by oxygen free radicals (O(2)(.-) and (.)OH in the absence of ethanol; O(2)(.-) and ethanol-derived peroxyl radicals, RO(2)(.), in the presence of ethanol) generated by gamma radiolysis. Initial radiolytic yields as determined by several markers of lipid peroxidation [i.e. decrease in endogenous antioxidants alpha-tocopherol and beta-carotene, formation of conjugated dienes and of thiobarbituric acid-reactive substances (TBARS)] were determined in 3 g liter(-1) LDLs (expressed as total LDL concentration) in the absence of ethanol or its presence at six different concentrations (0.42-17 x 10(-2) mol liter(-1)). Ethanol acted as an antioxidant by decreasing the rate of consumption of LDL endogenous antioxidants and the yields of formation of lipid peroxidation products, and by delaying the onset of the propagation phase for conjugated dienes and TBARS. With regard to the different markers studied, except for alpha-tocopherol and beta-carotene consumption, the effect of ethanol did not appear to be dependent on its concentration. Indeed, (.)OH were scavenged by ethanol at the lowest ethanol concentration (0.42 x 10(-2) mol liter(-1)), leading to RO(2)(.). These RO(2)(.) resulted in lower radiation-induced yields related to endogenous antioxidant consumption or to formation of lipid peroxidation products (for example, approximately 10% of RO(2)(.) oxidized LDLs from TBARS). Thus, under our in vitro conditions, ethanol behaved as an antioxidant when added to the LDL solutions. This should be taken into account in the reported antioxidant activity of wine. This is also of interest when lipophilic compounds have to be added as ethanolic solutions to LDLs to evaluate in vitro their antioxidant activity toward LDL peroxidation.

Determination of the yield of radiation-induced peroxidation of sodium linoleate in aqueous monomeric and micellar solutions.

Peroxidation of polyunsaturated fatty acids such as linoleic acid in aqueous micellar solution proceeds through a free-radical chain mechanism and is accompanied by the formation of conjugated dienes, some in the form of hydroperoxides. In the course of an investigation of radiation-induced oxidation of aqueous sodium linoleate, we have measured three indexes of peroxidation-conjugated dienes, hydroperoxides and thiobarbituric acid-reactive substances-by means of absorption spectroscopy, high-pressure liquid chromatography and spectrofluorimetry, respectively. There are linear correlations between the amounts of conjugated dienes, hydroperoxides and thiobarbituric acid-reactive substances. The radiolytic yields have been determined from the radiation dose dependence of the three markers of peroxidation as a function of sodium linoleate concentration. The results obtained indicate a strong effect of the concentrations of oxygen and linoleate on the yields of the products. The yields at different lipid concentrations display a large increase in chain propagation length; this is discussed in terms of the effect of micellar size.

Intramolecular semiquinone disproportionation in DNA. Pulse radiolysis study of the one-electron reduction of daunorubicin intercalated in DNA.

The one-electron reduction of daunorubicin, a quinonic antitumor antibiotic, intercalated in DNA was studied by pulse radiolysis using carboxyl radicals as reductants. The reaction's first stage is the daunorubicin semiquinone formation (k = 1.9 x 10(8) mol-1.dm3.s-1) in a way entirely consistent with a simple competition between .COO- disproportionation and the drug reduction. The semiquinone drug disappears by a first-order reaction (k = 1340 s-1) producing the hydroquinone form. This reaction leads to an equilibrium similar to the one without DNA and the equilibrium constant is very close to its value free in water (Kc approximately 25). In addition, the stoichiometry of the first-order reaction is the one of a dismutation process. Therefore, it appears that the disproportionation occurs along an intramolecular path across DNA. This migration takes place under our experimental conditions, over a distance of ca. 100 base pairs, with a mobility of ca. 4.4 X 10(-11) m2.V-1.s-1, similar in magnitude to an excess electron mobility in doped organic polymers.

One-electron reduction of daunorubicin intercalated in DNA or in a protein: a gamma radiolysis study.

The one-electron reduction of daunorubicin, an anthracycline antibiotic, intercalated in DNA or in the apoprotein of the riboflavin binding protein, was studied by gamma radiolysis. The two reduction mechanisms appear very similar to the one found for the non-intercalated drug. Hydrogen peroxide, which oxidizes non-intercalated hydroquinone daunorubicin with two electrons in one step (C. Houée-Levin, M. Gardès-Albert and C. Ferradin, FEBS lett., 173, 27-30, (1984)), reacts with daunorubicin hydroquinone in DNA but not in the protein. It appears thus that the site accessibility to hydrogen peroxide in DNA is better than in the protein. Biological consequences are discussed.

One-electron reduction of daunorubicin intercalated in DNA or in a protein: a gamma radiolysis study.

The one-electron reduction of daunorubicin, an anthracycline antibiotic, intercalated in DNA or in the apoprotein of the riboflavin binding protein, was studied by gamma radiolysis. The two reduction mechanisms appear very similar to the one found for the non-intercalated drug. Hydrogen peroxide, which oxidizes non-intercalated hydroquinone daunorubicin with two electrons in one step (C. Houée-Levin, M. Gardès-Albert and C. Ferradin, FEBS lett., 173, 27-30, (1984], reacts with daunorubicin hydroquinone in DNA but not in the protein. It appears thus that the site accessibility to hydrogen peroxide in DNA is better than in the protein. Biological consequences are discussed.

gamma-radiolysis study of the reduction by COO- free radicals of daunorubicin intercalated in DNA.

The reduction of daunorubicin intercalated in DNA was studied using COO- free radicals produced by gamma-radiolysis as reductants. The reduction process of the drug intercalated in DNA was found to be very similar to the one of daunorubicin in aqueous solution without DNA. (a) the final product is the same (7-deoxy daunomycinone); (b) the reduction yield is the same [2.6 +/- 0.2) x 10(-7) mol.J-1); (c) H2O2 reacts with hydroquinone daunorubicin giving back the drug in a one-step reaction. However 7-deoxy daunomycinone precipitation was so slow that this aglycone could be reduced by COO- free radicals giving its hydroquinone form, which cannot be observed without DNA. This shows that the whole 4-electron reduction process takes place in DNA. The implications of these findings are discussed.