In vivo interaction of carcinogenic acrylamide with cytochrome p450 isozymes and phase II enzymes in rabbit liver, kidney and lung

Nuyan, Mine
Acrylamide is an industrially produced chemical with known neurotoxic, reproductive toxin and carcinogenic effects. The carcinogenicity associated with acrylamide is mostly attributed to its metabolism by liver CYP2E1. However, studies investigating the effects of acrylamide on CYP2E1 enzyme are limited. In this study, it was aimed to investigate in vivo interaction of carcinogenic acrylamide on microsomal cytochrome P450 enzyme activities, and protein levels, and on cytosolic NQO1 and GST enzyme activities of rabbit liver, kidney and lung of acrylamide-treated rabbits. The in vivo protective effect of resveratrol, a phenolic compound, was also investigated on acrylamide toxicity. New Zealand male rabbits were treated with acrylamide and resveratrol, separately in different doses and conditions. Their combined effects were also investigated. CYP2E1-dependent p-Nitrophenol hydroxylase, NDMA N-demethylase and aniline 4-hydroxylase activities were found to be significantly increased in acrylamide-treated rabbit liver (1.80-3.0 fold) and kidney (1.6-fold). Rabbit liver and kidney CYP2E1 protein levels (determined by western blot analyisis) also increased approximately 2-fold due to acrylamide treatment. In rabbit liver, resveratrol was found significantly effective in decreasing both acrylamide-induced CYP2E1-dependent enzyme activities (approximately 1.5-1.80 fold) and CYP2E1 protein levels (approximately 1.5-1.70 fold). Additionally, resveratrol significantly decreased acrylamide-induced CYP2E1 protein level (2-2.5 fold) in rabbit kidney. However, no significant change was observed in rabbit lung CYP2E1-dependent enzyme activities and CYP2E1 protein levels due to acrylamide, resveratrol or their combined treatments. Furthermore, it was found that acrylamide treatment significantly increased CYP3A6-dependent erythromycin N-demethylase enzyme activity (1.85-fold) and CYP3A6 protein levels in rabbit liver (1.69-fold). No change was observed in CYP2B4-dependent benzphetamine N-demethylase enzyme activities of rabbit liver, kidney and lung by in vivo acrylamide, resveratrol or their combined treatments. Moreover, total GST and GST-Mu activities of rabbit kidney (1.5-fold, respectively) and total GST activity of rabbit lung (1.6-fold) were increased significantly only in resveratrol treated group. NQO1 enzyme activity of rabbit kidney was significantly increased by acrylamide treatment (1.6-fold). The results of the present study have demonstrated for the first time that acrylamide induces rabbit liver and kidney CYP2E1-dependent enzyme activities and CYP2E1 protein levels. The induction of CYP2E1 enzyme activity and protein level by acrylamide treatment can stimulate formation of other toxic compounds and procarcinogens metabolized by CYP2E1 which in turn further potentiates the risk of hepatotoxicity, mutagenicity and carcinogenicity. In the present study, it was also demonstrated for the first time that acrylamide treatment also increases CYP3A6 enzyme activity in rabbit liver which may lead to alterations in drug metabolism. The results of this study have also suggested that resveratrol may have protective effects on acrylamide induced toxicity; however, further in vivo studies are required to clarify the effect of resveratrol on both acrylamide-induced toxicity and anti-oxidant enzymes.


Investigation of sugar metabolism in rhizopus oryzae
Büyükkileci, Ali Oğuz; Hamamcı, Haluk; Department of Biotechnology (2007)
Rhizopus oryzae is a filamentous fungus, which can produce high amounts of L(+)-lactic acid and produces ethanol as the main by-product. In an effort to understand the pyruvate branch point of this organism, fermentations under different inoculum and glucose concentrations were carried out. At low inoculum size (1x103 spores ml-1), high amount of lactate (78 g l-1) was produced, whereas high ethanol concentration (37 g l-1) was obtained at high inoculum sizes (1x106 spores ml-1). Decreasing working volume i...
Production of an acetone-butanol-ethanol mixture from Clostridium acetobutylicum and its conversion to high-value biofuels
Sreekumar, Sanil; Baer, Zachary C.; Pazhamalai, Anbarasan; Günbaş, Emrullah Görkem; Grippo, Adam; Blanch, Harvey W.; Clark, Douglas S.; Toste, F. Dean (Springer Science and Business Media LLC, 2015-03-01)
Clostridium acetobutylicum is a bacterial species that ferments sugar to a mixture of organic solvents (acetone, butanol and ethanol). This protocol delineates a methodology to combine solventogenic clostridial fermentation and chemical catalysis via extractive fermentation for the production of biofuel blendstocks. Extractive fermentation of C. acetobutylicum is operated in fed-batch mode with a concentrated feed solution (500 grams per liter glucose and 50 grams per liter yeast extract) for 60 h, producin...
In vivo examination of the effects of hydroxycinnamic acid on xenobiotic metabolizing and antioxidant enzymes
SEMİZ, ASLI; Celik-Turgut, Gurbet; KARAKURT, SERDAR; AKÇA, HAKAN; ARSLAN, ŞEVKİ; Adalı, Orhan; ŞEN, ALAATTİN (National Library of Serbia, 2017-01-01)
In the last decade, hydroxycinnamic acids (HCA) have gained increasing attention from researchers due to their antioxidant potential. The aim of this study was to examine in detail the impact of dietary HCA on particular types of P450 and also selected phase II and antioxidant enzymes in Wistar rat. HCA (10 mu M/kg/day, i.p.) was administered for ten continuous days. Examination of the activities and mRNA and protein levels revealed that CYP2B, 2C6 and 3A enzyme activities were not altered significantly, wi...
The effect of cysteine-43 mutation on thermostability and kinetic properties of citrate synthase from Thermoplasma acidophilum
Kocabıyık, Semra; Russel, RJM; Danson, MJ; Hough, DW (Elsevier BV, 1996-07-05)
In this study, we have substituted serine-43 by cysteine in the recombinant citrate synthase from a moderately thermophilic Archaeon Thermoplasma acidophilum, for site-specific attachment of labels and have investigated the effects of this mutation on the biochemical properties and thermal stability of the enzyme. Both wild-type and the mutant enzymes were purified to homogenity using affinity chromatography on Matrex Gel Red A. The mutant Thermoplasma citrate synthase is very similar to wild-type citrate s...
In vitro activity of panomycocin, a novel exo-beta-1,3-glucanase isolated from Pichia anomala NCYC 434, against dermatophytes
Izgu, Fatih; Altinbay, Demet; Tureli, Akif Emre (2007-01-01)
Killer proteins that are produced and secreted into the environment by certain yeast strains are considered as promising antifungal agents. In this study, in vitro activity of Pichia anomala NCYC 434 (K5) killer protein, panomycocin, which is a 49 kDa glycoprotein with an exo-beta-1,3-glucanase activity was tested against 41 isolates of dermatophytes. Minimum inhibitory concentrations (MICs) were determined by a broth microdilution method based on the reference document M38-A of Clinical and Laboratory Stan...
Citation Formats
M. Nuyan, “In vivo interaction of carcinogenic acrylamide with cytochrome p450 isozymes and phase II enzymes in rabbit liver, kidney and lung,” M.S. - Master of Science, Middle East Technical University, 2008.