Modulation by certain factors of metabolic activation of aflatoxin B1 as detected in vitro in a simple fluorimetric assay.
Forty compounds belonging to various chemical groups have been tested for their ability to suppress metabolic activation of aflatoxin B1 (AFB1) mediated by rat liver microsome. Microsomal activation has been carried out in an in vitro system containing Tris-buffer at pH 7.2. Production of the reactive metabolite, AFB1 8,9-epoxide, has been measured by separation and detection of its hydrolysis product AFB1 8,9-dihydrodiol as the Tris-diol complex. The complex is separated using simple procedures of biphasic extraction and deproteinization, and detected by its characteristic fluorescence. Quantitation is made by direct comparison of its fluorescence with that of an authentic Tris-diol prepared synthetically. The method is rapid and proved to be highly sensitive and reproducible. A large number of compounds have been observed to modulate at varying degrees the activation of AFB1 in this in vitro system. Many compounds have been tested at several concentration ranges and inhibition curve is constructed in each case from which ID50 values, i.e., the dose needed to bring about 50% inhibition can be obtained. These values expressed as nmol afford a direct and realistic comparison of the inhibitory potential of various modulators. Factors having great inhibitory potential have been identified as retinoids (retinol, retinal, retinoic acid, retinyl acetate), beta-carotene, riboflavin, ascorbic acid, copper, zinc, linoleic acid, p-hydroxy benzoic acid, butylated hydroxyanisole, butylated hydroxytoluene, disulfiram, and phenothiazine. Several other compounds have shown moderate inhibitory potential. The strong inhibition on Tris-diol formation by several vitamins, antioxidants and trace metals shows similarity with their effect on AFB1-DNA adduct formation. It is suggested that these agents may have potential anticarcinogenic activity against AFB1.
D016604:Aflatoxin B1; D000348:Aflatoxins / Q000378:metabolism*; D000818:Animals; D001711:Biotransformation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D000348:Aflatoxins; D016604:Aflatoxin B1