OBJECTIVES: 17α-Ethinylestradiol (EE2) is an endocrine disruptor that is an ingredient of oral contraceptives. Here, EE2 metabolism catalyzed by cytochromes P450 (CYP) was studied. Two model organisms, rat and ligninolytic fungus Pleurotus ostreatus, were used. METHODS: To resolve the role of rat and/or fungal CYPs in EE2 oxidation, microsomes were incubated with EE2 and NADPH or cumene hydroperoxide. Using Supersomes™, we examined which of rat CYPs oxidize EE2. RESULTS: EE2 is effectively degraded by P. ostreatus in vivo. In vitro, EE2 is metabolized by CYPs by the NADPH-dependent and organic hydroperoxide-dependent mechanisms. Rat hepatic microsomes metabolize EE2 in the presence of NADPH to three products; two of them are hydroxylated EE2 derivatives. Using rat Supersomes™ we found that EE2 is hydroxylated by several rat CYPs, among them CYP2C6 and 2C11 are most efficient in 2-hydroxy-EE2 formation, while CYP2A and 3A catalyze EE2 hydroxylation to the second product. On the contrary, the products of the NADPH-dependent hydroxylating reactions were not detected in Pleurotus ostreatus. During the reaction of EE2 in microsomes isolated from rat and P. ostreatus in the presence of the alternate oxidant, cumene hydroperoxide, another metabolite, different from the above mentioned products, is generated. Rat CYP1A1 is the most efficient enzyme catalyzing formation of this EE2 product. CONCLUSION: The results suggest that CYPs play a role in EE2 metabolism in rat and P. ostreatus. To our knowledge this is the first finding describing ligninolythic fungal metabolism of EE2 by CYP in the presence of cumene hydroperoxide.

• MeSH
• aromatické hydroxylasy metabolismus   MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• cytochrom P-450 CYP3A metabolismus   MeSH
• ethinylestradiol metabolismus   MeSH
• hydroxylace MeSH
• jaterní mikrozomy enzymologie   metabolismus   MeSH
• krysa rodu rattus MeSH
• oxidace-redukce MeSH
• Pleurotus MeSH
• potkani Wistar MeSH
• steroid-16-alfa-hydroxylasa metabolismus   MeSH
• steroid-21-hydroxylasa metabolismus   MeSH
• steroidhydroxylasy metabolismus   MeSH
• systém (enzymů) cytochromů P-450 metabolismus   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim was to investigate whether rosuvastatin affect rat cytochrome P450 (CYP) 2C11 and CYP2C6. CYP2C11 and CYP2C6 are considered as counterparts of human CYP2C9, which metabolizes many drugs including S-warfarin, diclofenac or ibuprofen. The male Wistar rats were fed standard laboratory diet (STD) or high cholesterol diet (HCD, 1% of cholesterol, 10% of lard fat) for 21 days. Rosuvastatin administration in STD (0.03% w/w) resulted in decreased mRNA expression of CYP2C11 as well as of CYP2C6 (here significant) and in a significant decrease of the respective protein expression as well as of the enzyme activity of both CYP2C forms. When rosuvastatin was administered in the HCD, the mRNA expression of both CYP2C forms was significantly lowered; the protein and activity parameters did not show significant changes. These results suggest that CYP2C11 as well as CYP2C6 expression and activity are negatively affected by rosuvastatin and may be modulated by high cholesterol high fat diet. Therefore, it should be taken into consideration that drugs metabolized by CYP2C9 in human could interact with rosuvastatin, as it has been already suggested for warfarin (rosuvastatin has increased its anticoagulant effect in human), and for telmisartan, sildenafil and glimepiride.

• MeSH
• aromatické hydroxylasy genetika   metabolismus   MeSH
• dieta s vysokým obsahem tuků MeSH
• fluorbenzeny aplikace a dávkování   farmakologie   MeSH
• jaterní mikrozomy účinky léků   enzymologie   MeSH
• krysa rodu rattus MeSH
• lipidy krev   MeSH
• potkani Wistar MeSH
• pyrimidiny aplikace a dávkování   farmakologie   MeSH
• regulace genové exprese enzymů účinky léků   MeSH
• steroid-16-alfa-hydroxylasa genetika   metabolismus   MeSH
• steroid-21-hydroxylasa genetika   metabolismus   MeSH
• sulfonamidy aplikace a dávkování   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

This short communication is aimed to investigate whether the widely used hypolipidemic drug fenofibrate affects CYP2C11 and CYP2C6 in rats, both counterparts of human CYP2C9, known to metabolise many drugs including S-warfarin and largely used non-steroidal antiinflammatory drugs such as ibuprofen, diclofenac and others. The effects of fenofibrate on the expression of rat liver CYP2C11 and CYP2C6 were studied in both healthy Wistar rats and hereditary hypertriglyceridemic rats. Both strains of rats were fed on diet containing fenofibrate (0.1% w/w) for 20 days. Fenofibrate highly significantly suppressed the expression of mRNA of CYP2C11 and less that of CYP2C6 in liver microsomes of both rat strains; this effect was associated with a corresponding decrease in protein levels. The results indicate that the combination of fenofibrate with drugs metabolised by CYP2C9 in humans should be taken with caution as it may lead, for example, to the potentiation of warfarin effects. This type of drug interaction has been observed previously and the results presented here could contribute to the explanation of their mechanism.

• MeSH
• aromatické hydroxylasy biosyntéza   MeSH
• enzymová represe MeSH
• fenofibrát farmakologie   MeSH
• hypertriglyceridemie metabolismus   MeSH
• hypolipidemika farmakologie   MeSH
• jaterní mikrozomy účinky léků   metabolismus   MeSH
• krysa rodu rattus MeSH
• potkani Wistar MeSH
• steroid-16-alfa-hydroxylasa biosyntéza   MeSH
• steroid-21-hydroxylasa biosyntéza   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

OBJECTIVES: The aim of the study was to find whether probiotic Lactobacillus casei influences the expression or the activity of cytochromes P450 (CYP) and whether it has an influence on the level of CYP mRNA in male rats. DESIGN: Live bacterial suspension of L. casei was administered orally (gavage) to healthy male Wistar rats daily for 7 days. Control group of rats was treated with the saline solution. Sections of the duodenum, jejunum, ileum, caecum and colon were dissected from each experimental animal. In all individual samples, the expression of selected CYPs was determined by Western blotting. The levels of expression of CYPs were also evaluated by mRNA using the real-time PCR method. RESULTS: There were changes observed in the expression of CYP enzymes and in the CYP mRNA levels along the intestine after application of L. casei. The expression of CYP1A1 enzyme was found to be decreased in the proximal part of the jejunum and colon, CYP1A1 mRNA level was decreased in the distal part of the jejunum, ileum and caecum. Thus, the changes in CYP1A1 protein or mRNA were observed along the intestine of male rats. Similarly, a decreased expression of the caecal CYP2E1 mRNA and of the duodenal CYP3A9 mRNA after treatment of rats with L. casei was found. CONCLUSION: Probiotic L. casei might be able to contribute to prevention against colorectal cancer by decreasing levels of certain forms of xenobiotic-metabolizing enzymes; moreover, in general, there is a possibility of interactions with concomitantly taken pharmacotherapeutic agents.

• MeSH
• aktivace enzymů účinky léků   MeSH
• aromatické hydroxylasy genetika   metabolismus   MeSH
• cytochrom P-450 CYP2E1 genetika   metabolismus   MeSH
• cytochrom P-450 CYP3A genetika   metabolismus   MeSH
• cytochromy genetika   metabolismus   MeSH
• játra účinky léků   metabolismus   mikrobiologie   MeSH
• krysa rodu rattus MeSH
• Lactobacillus casei fyziologie   MeSH
• messenger RNA metabolismus   MeSH
• potkani Wistar MeSH
• probiotika farmakologie   MeSH
• regulace genové exprese enzymů účinky léků   MeSH
• steroid-16-alfa-hydroxylasa genetika   metabolismus   MeSH
• steroid-21-hydroxylasa genetika   metabolismus   MeSH
• střeva účinky léků   metabolismus   mikrobiologie   MeSH
• systém (enzymů) cytochromů P-450 genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

A new, simple, rapid, sensitive, and repeatable reversed-phase HPLC method was developed and validated for the simultaneous determination of tolbutamide, phenacetin and their metabolites in rat liver perfusate. Chlorpropamide was used as an internal standard to ensure the precision and accuracy of this method. Analytes were extracted into diethyl ether using a two-step liquid-liquid extraction. A C18 analytical column and a mobile phase composed of acetonitrile and potassium phosphate buffer were used for the chromatographic separation with UV detection. Limits of detection varied between 20 and 46ng/mL for phenacetin, tolbutamide and their metabolites. The overall extraction recovery for the analytes varied from 65.4% in paracetamol to 88.0% in tolbutamide for concentrations within the expected range of concentrations from previous experimental samples. In terms of precision, the intra- and inter-day variation at three different concentrations in all analytes never exceeded 7.6 and 11.4%, respectively. This method is applicable for the modeling and description of possible pharmacological interactions on rat cytochromes P450 1A2 and 2C6/11 or can be used for in vitro evaluation of both cytochromes 1A2 and 2C.

• MeSH
• aromatické hydroxylasy metabolismus   MeSH
• biologické markery metabolismus   MeSH
• biologické modely MeSH
• cytochrom P-450 CYP1A2 metabolismus   MeSH
• fenacetin metabolismus   MeSH
• játra enzymologie   MeSH
• krysa rodu rattus MeSH
• lékové interakce fyziologie   MeSH
• perfuze metody   MeSH
• potkani Wistar MeSH
• steroid-16-alfa-hydroxylasa metabolismus   MeSH
• steroid-21-hydroxylasa metabolismus   MeSH
• tolbutamid metabolismus   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH