Beta carotene and its oxidation products have different effects on microsome mediated binding of benzo[a]pyrene to DNA.
The effects of beta-carotene (betaC) and its oxidation products on the binding of benzo[a]pyrene (BaP) metabolites to calf thymus DNA was investigated in the presence of rat liver microsomes. Mixtures of betaC oxidation products (betaCOP) as well as separated, individual betaC oxidation products were studied. One set of experiments, for example, involved the use of the mixture of betaCOP obtained after a 2-h radical-initiated oxidation. For this data set, the incorporation of unoxidized betaC into microsomal membranes caused the level of binding of BaP metabolites to DNA to decrease by 29% over that observed in the absence of betaC; however, the incorporation of the mixture of betaCOP caused the binding of BaP metabolites to DNA to increase 1.7-fold relative to controls without betaC. Two variations of this experiment were studied: (1) When no NADPH was added, betaC decreased the binding of BaP metabolites to DNA by 19%, but the mixture of betaCOP increased binding by 3.3-fold relative to that observed in the absence of betaC. (2) When NADPH was added under near-anaerobic conditions, betaC caused an almost total (94%) decrease in binding whereas betaCOP had no effect on the amount of binding relative to that observed in the absence of betaC. Both betaCOP and cumene hydroperoxide caused BaP metabolites to bind to DNA even when NADPH was omitted from the incubation mixture. Separation of the mixture of betaC oxidation products into fractions by HPLC allowed preliminary testing of individual betaC oxidation products separately; of the various fractions tested, the products tentatively identified as 11,15'-cyclo-12,15-epoxy-11,12,15,15'-tetrahydro-beta-carotene and beta-carotene-5,6-epoxide appeared to cause the largest increase in BaP-DNA binding. Microsomes from rats induced with 3-methylcholanthrene (3MC) or Aroclor 1254 produced different levels of binding in some experimental conditions. We hypothesize that, under some conditions, the incorporation of betaC into microsomal membranes can be protective against P450-catalyzed BaP binding to DNA; however, the incorporation of betaCOP facilitates the formation of BaP metabolites that bind DNA, although only certain P450 isoforms catalyze the binding process.
Salgo
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MG
The Biodynamics Institute, Louisiana State University, Baton Rouge 70803-1800, USA.
Cueto
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R
Winston
G W
GW
Pryor
W A
WA
D000818:Animals; D001564:Benzo(a)pyrene / Q000378:metabolism*; D002417:Cattle; D020111:Chlorodiphenyl (54% Chlorine) / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D018736:DNA Adducts / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D013053:Spectrophotometry; D019207:beta Carotene / Q000378:metabolism / Q000494:pharmacology*
D018736:DNA Adducts; C056612:benzo(a)pyrene-DNA adduct; D019207:beta Carotene; D020111:Chlorodiphenyl (54% Chlorine); D001564:Benzo(a)pyrene; D009249:NADP; D008748:Methylcholanthrene; D004247:DNA; D003577:Cytochrome P-450 Enzyme System