Modification of hepatic drug-metabolizing enzymes in rats treated with alkyl sulfides
Introduction
Allium consumption has been associated in epidemiologic studies with a reduced risk of cancer incidence 5, 6, 28, 34. The anticarcinogenic properties of Allium may be attributed, at least in part, to organosulfur compounds derived from these plants. Experimental studies on animal models have shown an inhibition of chemically induced carcinogenesis in different organs by certain sulfur-containing compounds 13, 14, 15, 16, 20, 27, 31, 32. Many investigators hypothesized that chemopreventive agents inhibiting carcinogenesis may act by altering the metabolism of procarcinogen. These agents can increase detoxification by increasing the levels of phase II enzymes such as glutathione S-transferase (GST), UDP-glucuronyl transferase (UGT) or quinone reductase (QR) or by decreasing the levels of some phase I enzymes such as cytochromes P450 10, 33. Allyl sulfides derived from garlic have been shown to increase phase II enzymes such as GST, UGT and epoxide hydrolase (EH) 9, 11, 17, 27and to inhibit cytochrome P450 2E1 3, 4, 18, 24, 29. All these studies have focused on the effects of diallyl sulfide (DAS) and diallyl disulfide (DADS), whereas other sulfur compounds of Allium species have been investigated to a lesser degree.
In the present work we have investigated the ability of some alkyl sulfides to modulate in vivo a variety of drug-metabolizing enzymes in the liver of male Wistar rats. Alkyl sulfides are constituents of Allium species, generated from highly reactive sulfenic acids which are released from various naturally occurring S-alk(en)yl-l-cysteine sulfoxides by the action of the enzyme alliinase [1]. Propyl groups are present in onion, shallot, leek and scallion but are absent in garlic, whereas allyl groups predominate in garlic and methyl groups predominate in Chinese chive [2]. Dimethyl sulfide (DMS), dimethyl disulfide (DMDS), methylpropyl disulfide (MPDS), dipropyl sulfide (DPS) and dipropyl disulfide (DPDS) were selected in this study (Fig. 1). The effects of alkyl sulfides were compared to those of DADS which were studied previously [11]and to the classical P450 2B inducer, phenobarbital (PB). Marker activities of the P450 isoenzymes that are important in carcinogen metabolism have been measured in liver microsomes, namely ethoxyresorufin O-deethylase (EROD), a marker of CYP 1A1, methoxyresorufin O-demethylase (MROD), a marker of CYP 1A2, pentoxyresorufin O-dealkylase (PROD), a marker of CYP 2B1,2, nitrosodimethylamine N-demethylase (NDMAD), a marker of CYP 2E1 and erythromycin N-demethylase (ERDM), a marker of CYP 3A. Immunoblots were performed to assess the microsomal levels of these isoenzymes (CYP 1A1, CYP 1A2, CYP 2B1, CYP 2B2 and CYP 2E1). Phase II enzyme activities were also assayed including UGTs, GST and QR.
Section snippets
Chemicals
DMS (purity 99%), DMDS (purity 98%), MPDS (purity 95%), DPS (purity 97%), DPDS (purity 98%) and DADS (purity 80%, remainder other allyl sulfides) were obtained from Aldrich. They were used without further purification. PB (sodium salt) was purchased from Coopérative Pharmaceutique Francaise, Melun, France. Polyclonal antibodies anti-P450 1A and anti-P450 2B were a gift of Professor A.-M. Batt (Centre du Médicament, Nancy, France) and polyclonal antibody anti-P450 2E1 was a gift of Professor P.
Results
Food consumption was the same in all groups during the experiment (data not shown) as were body weights at the end of the experiment (Table 1). Relative liver weights were significantly increased by DPS (10%), DADS (45%) and PB (20%) (Table 1). The increase was greatest with DADS. Total hepatic P450 was increased by DPS (70%), DPDS (30%), DADS (36%) and PB (110%). NADPH-cytochrome P450 reductase activity was significantly stimulated by DPS and PB treatments.
The effects of the treatments on
Discussion
The present study shows that the administration of alkyl or allyl sulfides to rats modulates activities of drug-metabolizing enzymes. Compounds containing methyl groups had negligible effect, whereas those containing two propyl groups or two allyl groups provoked a pleiotropic response on drug-metabolizing enzymes. This response is similar to the one produced by the prototypical inducer PB.
DPS, DPDS and DADS, administered by gavage, induced different monooxygenase activities, i.e. EROD, MROD
Acknowledgements
We thank Marie-France Vernevaut (INRA, Dijon, France) for excellent technical assistance and Joël Leclerc (INRA, Dijon, France) for the determination of enzyme activities and statistic calculations. Antibodies anti-P450 1A and anti-P450 2B were kindly given by Professor A.-M. Batt and anti-P450 2E1 by Professor P. Beaune. This work was supported by a grant of Conseil Régional de Bourgogne and Institut National de la Recherche Agronomique, France.
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