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
OBJECTIVES: The aim of the study was to find whether probiotic Escherichia coli Nissle 1917 O6:K5:H1 (EcN) influences the expression of cytochromes P450 (CYP) in the rat intestine. DESIGN: Live bacterial suspension of EcN was administered to healthy male Wistar rats daily for 7 days. Control group of rats was stressed by oral application of the saline solution daily for 7 days as well. Sections of the duodenum, jejunum, ileum, caecum and colon have been taken from each experimental animal. With all individual samples, microsomal fraction has been prepared and expression of selected CYPs was determined by Western blotting. The levels of expression of CYPs were also evaluated by mRNA using real-time PCR. RESULTS: It was found that there are changes in expression of CYP enzymes studied along the intestine. CYP1A1, 2B1/2 and 2E1 are present mainly in the duodenum and jejunum; on the other hand, CYP2C6 is expressed mainly in the caecum and colon. CYP3A was found all over the rat intestine. The results show that there are no prominent differences between control samples and samples with EcN, only the expression of CYP3A protein in the duodenum appears to exhibit a clear tendency to decrease. In the case of the colon, a significant increase in the expression of CYP3A (most likely CYP3A1) after treatment of rats with EcN was found. CONCLUSION: This in vivo study revealed that the levels of colon CYP3A could be significantly increased in rats treated with probiotic EcN. On the contrary, the expression of CYP3A in the duodenum decreased. However, the changes in the expression of CYP enzymes are probably not as extensive to be clinically important in man; hence, most likely the probiotic EcN has little influence on the intestinal drug metabolism by CYP enzymes.
- MeSH
- aromatické hydroxylasy metabolismus MeSH
- cytochrom P-450 CYP3A metabolismus MeSH
- duodenum enzymologie MeSH
- Escherichia coli MeSH
- gastrointestinální trakt enzymologie MeSH
- kolon enzymologie MeSH
- krysa rodu rattus MeSH
- modely u zvířat MeSH
- potkani Wistar MeSH
- probiotika farmakologie MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cílem in vitro studie bylo získat informace o možnosti interakce rokuronia s lidským jaterním mikrosomálním systémem cytochromů P450, zejména s formou cytochromu P450 3A4. Metody: Interakce rokuronia s mikrosomálními cytochromy P450 byla sledována spektrofotometricky pomocí diferenční absorpční spektroskopie. Interakce s cytochromem P450, formou CYP3A4, byla sledována pomocí inhibice specifické aktivity CYP3A4 (testosteron 6ß-hydroxylace). Výsledky: Rokuronium interaguje in vitro s lidskými jaterními mikrosomálními cytochromy P450 a inhibuje účinně aktivitu formy CYP3A4. Závěr: Vzhledem k tomu, že tato forma metabolizuje látky steroidní povahy, je možný vliv interakce rokuronia na další procesy v organizmu. Nelze rovněž vyloučit i možnost lékových interakcí rokuronia s léčivy metabolizovanými CYP3A4, které by mohly např. vést k prodloužení účinku rokuronia.
Aim of the study was to find whether human liver microsomal cytochromes P450 interact with rocuronium in vitro. Methods: Interaction of rocuronium with microsomal cytochromes P450 was followed spectrophotometrically using difference absorption spectroscopy. Interaction with cytochrome P450 form CYP3A4 was studied by determination of inhibition of a CYP3A4-specific enzyme activity (testosterone 6ß-hydroxylation) by HPLC. Results: Rocuronium interacts in vitro with human liver microsomal cytochromes P450 and inhibits effectively the CYP3A4 specific enzyme activity. Conclusion: Due to the fact that the CYP3A4 form is known to metabolize steroids and steroid-related compounds there is a possibility of an influence of rocuronium-CYP3A4 interactions on other processes in the organism. Moreover, a possibility of drug interactions with concomitantly administered drugs metabolized by CYP3A4 leading to e.g. prolongation of rocuronium effect cannot be excluded.
- MeSH
- androstanoly farmakokinetika farmakologie metabolismus MeSH
- financování organizované MeSH
- inhibitory cytochromu P450 MeSH
- jaterní mikrozomy enzymologie metabolismus MeSH
- nedepolarizující myorelaxancia farmakokinetika metabolismus MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- techniky in vitro MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
OBJECTIVES: The aim of the study was to find, whether probiotic Escherichia coli Nissle 1917 O6:K5:H1 (EcN) influence the amount and activity of cytochromes P450 (CYP) in rat liver. DESIGN: Live bacterial suspension of EcN was applied to the female Wistar rats in single dose or for 14 days consecutively. The bacterial lipopolysaccharide (LPS) isolated by phenol extraction from the EcN was given to the rats for 14 days as well. Control rats were treated with the saline solution daily for 14 days. Relative amount of CYP2C6, CYP2C9 (corresponding to rat CYP2C11), CYP3A1 and CYP1A2 protein expression in rat liver microsomes was determined by Western blotting. For the determination of six CYP activities (corresponding to human CYP1A2, CYP2A6, CYP2B6, CYP3A4, CYP2C9 and CYP2D6) fluorescence, luminescence or absorbance detection was used. RESULTS: The data presented show that the changes of the total content of the CYP enzymes in rat liver are not significant after administration of the probiotic for 1 or 14 days as well as of the LPS. Western blots revealed a slight increase in CYP2C6 protein expression; level of another rat CYP2C protein (readings with anti-human CYP2C9 antibody corresponding to the rat CYP2C11) as well as of CYP1A2 was elevated after administration of LPS; a small decrease was observed with CYP3A1 protein. Changes in activities of CYP forms are not significant, only the activity of rat CYP2C forms in liver microsomal samples of rats given free LPS appeared to exhibit a small, but significant tendency to increase. CONCLUSION: The results show that the p.o. administration of probiotics to rat does not markedly influence the rat hepatic CYP enzymes.
- MeSH
- časové faktory MeSH
- játra enzymologie metabolismus MeSH
- krysa rodu rattus MeSH
- lipopolysacharidy metabolismus MeSH
- potkani Wistar MeSH
- probiotika metabolismus MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
OBJECTIVES: To study the contribution of individual purified porcine CYP1A2, 2E1 and 2A19 enzymes to the biotransformation of skatole. METHODS: Individual porcine and human enzymes (CYP1A2, 2E1 or 2A6/19) were used to study their potential involvement in skatole metabolism. Furthermore, the inhibition experiments using specific inhibitors of CYP1A2, 2E1 or 2A6/19, were performed. For determination of skatole biotransformation by individual CYP forms in reconstituted systems, HPLC method with UV detection was used. RESULTS: The data presented in this paper show that porcine and human CYPs are responsible for the formation of indole-3-carbinol and 3-methyloxindole. Whereas in pig CYP2A19 and CYP1A2 seem to be the most important for metabolism of skatole, in man CYP1A2 and CYP2E1 forms are mainly responsible for the production of the metabolites mentioned above. CONCLUSIONS: The porcine and human CYP1A2, 2E1, 2A6/19 forms contribute to formation of 3-methyloxindole and indole-3-carbinol.
- MeSH
- cytochrom P-450 CYP1A2 metabolismus MeSH
- cytochrom P-450 CYP2E1 metabolismus MeSH
- indoly metabolismus MeSH
- inhibitory cytochromu P450 CYP1A2 MeSH
- inhibitory cytochromu P450 CYP2E1 MeSH
- inhibitory cytochromu P450 MeSH
- lidé MeSH
- prasata MeSH
- skatol metabolismus MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cytochromes P450 comprise a superfamily of proteins that play a crucial role in the biotransformation of numerous chemicals. Purified CYPs can be used e.g. in studies on structure or determining the drug metabolism pathways. In this work, purification of the porcine CYP1A and CYP2A19 to electrophoretic homogeneity from the pig hepatic microsomes using octylamino Sepharose and hydroxylapatite column chromatography is reported. The proteins have been clearly recognized by commercial antibodies against rat and human CYP1A2 (porcine CYP1A) and human CYP2A6 (CYP2A19) respectively, using Western blot. Activities of both enzymes were determined by specific substrates, 7-ethoxyresorufin, 7-methoxyresorufin (CYP1A) and coumarin (CYP2A19). The isolated enzymes show kinetic parameters similar to human counterparts. Taken together, pig cytochromes can be used for the testing of veterinary drug metabolism, useful for the determination of drug residues in meat of pigs. The results obtained show that the pigs may be a suitable model for biotransformation of xenobiotics in humans.
- MeSH
- aromatické hydroxylasy izolace a purifikace MeSH
- chromatografie agarózová MeSH
- cytochrom P-450 CYP1A1 izolace a purifikace MeSH
- jaterní mikrozomy enzymologie MeSH
- lidé MeSH
- prasata MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
