Formation of N-7-(2-carbamoyl-2-hydroxyethyl)guanine in DNA of the mouse and the rat following intraperitoneal administration of [14C]acrylamide.
ABSTRACT Acrylamide is an alkylating agent which reacts very slowly in direct reactions with DNA and is negative in the Ames test, but is carcinogenic in mice and rats. In order to explain the cancer-initiating properties of acrylamide we have studied DNA adduct formation in vitro with a metabolizing system and in vivo in mice and rats following i.p. administration of 14C-labeled acrylamide. A major adduct found in both species was N-7-(2-carbamoyl-2-hydroxy-ethyl)guanine, formed by reaction of the DNA with the epoxide metabolite glycidamide. The levels of this adduct were similar in the different organs of the two rodent species, which supports the notion that glycidamide is relatively evenly distributed among tissues and that the organ-specificity in acrylamide carcinogenesis cannot be explained by a selective accumulation of the DNA-reactive metabolite in target organs.
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ABSTRACT: Acrylamide is a toxin that humans are readily exposed to due to its formation in many carbohydrate rich foods cooked at high temperatures. Acrylamide is carcinogenic, neurotoxic and causes reproductive toxicity when high levels of exposure are reached in mice and rats. Acrylamide induced effects on fertility occur predominantly in males. Acrylamide exerts its reproductive toxicity via its metabolite glycidamide, a product which is only formed via the cytochrome P450 detoxifying enzyme CYP2E1. Glycidamide is highly reactive and forms adducts with DNA. Chronic low dose acrylamide exposure in mice relevant to human exposure levels results in significantly increased levels of DNA damage in terms of glycidamide adducts in spermatocytes, the specific germ cell stage where Cyp2e1 is expressed. Since cells in the later stages of spermatogenesis are unable to undergo DNA repair, and this level of acrylamide exposure causes no reduction in fertility, there is potential for this damage to persist until sperm maturation and fertilisation. Cyp2e1 is also present within epididymal cells, allowing for transiting spermatozoa to be exposed to glycidamide. This could have consequences for future generations in terms of predisposition to diseases such as cancer, with growing indications that paternal DNA damage can be propagated across multiple generations. Since glycidamide is the major contributor to DNA damage, a mechanism for preventing these effects is inhibiting the function of Cyp2e1. Resveratrol is an example of an inhibitor of Cyp2e1 which has shown success in reducing damage caused by acrylamide treatment in mice. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 04/2015; 777. DOI:10.1016/j.mrfmmm.2015.04.008 · 4.44 Impact Factor
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ABSTRACT: Acrylamide (AA) is reported present in high-temperature-processed food and classified as a possible human carcinogen. In vivo metabolic activation of AA by CYP 2E1 to glycidamide (GA) may play an important role on AA carcinogenicity. AA and GA can be detoxified by glutathione-S-transferase to form AA and isomeric GA glutathione conjugates (AA-, GA2- and GA3-GSH, respectively), which can be further metabolized to mercapturic acids (MAs). Although many studies analyzed MAs in urine of rodents and humans, few studies have characterized and analyzed the GSH conjugates. The objectives of this study were to synthesize, purify, and characterize AA-GSH, GA2-GSH, GA3-GSH, ((13)C3)-AA-GSH, ((13)C3)-GA2-GSH, and ((13)C3)-GA3-GSH to develop an isotope-dilution liquid chromatography tandem mass spectrometry (LC-MS/MS) method to analyze AA- and GA-GSHs in blood of rats treated with AA. After purification and characterization of these conjugates, the LC-MS/MS method was developed and validated. This method reveals a limit of detection (S/N = 3) at 0.017 and a limit of quantitation (S/N=10) at 0.05 ng/mL of serum for AA-GSH, 0.075 and 0.25 ng/mL for GA2-GSH, and 0.15 and 0.5 ng/mL for GA3-GSH. Analyzed with this method, AA-GSH, GA2-GSH and GA3-GSH were 1651.1 ± 374.5, 18.4 ± 6.3 and 75.3 ± 31.3 ng/mL in blood of male rats at 2 hr after treatment with 5 mg/kg bw of AA by ip injection. These results showed that the LC-MS/MS method was successfully developed to analyze AA-GSH, GA2-GSH and GA3-GSH with satisfying sensitivity of AA and GA which were conjugated by glutathione in vivo. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.Chemico-biological interactions 07/2015; 237:38-46. DOI:10.1016/j.cbi.2015.05.002 · 2.98 Impact Factor
Chiang Mai University Journal of Natural Sciences 01/2014; 13(1). DOI:10.12982/cmujns.2014.0016