The gut microbiota provides a wide range of beneficial functions for the host, and has an immense effect on the host's health status. The presence of microbiome in the gut may often influence the effect of an orally administered drug. Molecular mechanisms of this process are however mostly unclear. We investigated how the effect of a nonsteroidal drug nabumetone on expression of drug metabolizing enzymes (DMEs) in mice intestine and liver is changed by the presence of microbiota, here, using the germ free (GF) and specific pathogen free (SPF) BALB/c mice. First, we have found in a preliminary experiment that in the GF mice there is a tendency to increase bioavailability of the active form of nabumetone, which we have found now to be possibly influenced by differences in expression of DMEs in the GF and SPF mice. Indeed, we have observed that the expression of the most of selected cytochromes P450 (CYPs) was significantly changed in the small intestine of GF mice compared to the SPF ones. Moreover, orally administered nabumetone itself altered the expression of some CYPs and above all, in different ways in the GF and SPF mice. In the GF mice, the expression of the DMEs (CYP1A) responsible for the formation of active form of the drug are significantly increased in the small intestine and liver after nabumetone application. These results highlight the importance of gut microbiome in processes involved in drug metabolism in the both gastrointestinal tract and in the liver with possible clinical relevance.

• MeSH
• antiflogistika nesteroidní aplikace a dávkování   metabolismus   MeSH
• aplikace orální MeSH
• Bacteria metabolismus   MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• dysbióza MeSH
• játra účinky léků   enzymologie   MeSH
• metabolická aktivace MeSH
• myši inbrední BALB C MeSH
• nabumeton aplikace a dávkování   metabolismus   MeSH
• střevní mikroflóra * MeSH
• systém (enzymů) cytochromů P-450 genetika   metabolismus   MeSH
• tenké střevo účinky léků   enzymologie   mikrobiologie   MeSH
• transkripční faktory genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

1. Sesquiterpenes, constituents of plant essential oil, are popular bioactive compounds due to the positive effect on human health, but their potential toxicity and possible herb-drug interactions are often omitted. In our in vivo study, we followed up the effect of p.o. administration of two sesquiterpenes β-caryophyllene oxide (CAO) and trans-nerolidol (NER) on various xenobiotic-metabolizing enzymes in mice liver and small intestine. 2. To spot the early effect of studied compounds, enzymatic activity and mRNA levels were assessed 6 and 24 h after single dose. 3. CAO and NER markedly increased cytochromes P450 (CYP2B, 3A, 2C) activity and mRNA levels in both tissues. Liver also showed elevated activity of aldo-ketoreductase 1C and carbonyl reductase after treatment. Contrary, sesquiterpenes decreased NAD(P)H:quinone oxidoreductase 1 activity in small intestine. Among conjugation enzymes, only liver sulfotransferase activity was increased by sesquiterpenes. 4. Our results document that single dose of sesquiterpenes modulate activities and expression of several xenobiotic-metabolizing enzymes.

• MeSH
• aldehydreduktasa metabolismus   MeSH
• enzymy metabolismus   MeSH
• estradioldehydrogenasy metabolismus   MeSH
• inbrední kmeny myší MeSH
• játra účinky léků   enzymologie   MeSH
• metabolická inaktivace účinky léků   MeSH
• NAD(P)H dehydrogenasa (chinon) metabolismus   MeSH
• regulace genové exprese enzymů účinky léků   MeSH
• seskviterpeny farmakologie   toxicita   MeSH
• systém (enzymů) cytochromů P-450 genetika   metabolismus   MeSH
• tenké střevo účinky léků   enzymologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The prevalence of obesity is rapidly increasing across the world. Physiologic alterations associated with obesity are known to alter enzyme expression and/or activities. As drug-metabolizing and antioxidant enzymes serve as defense system against potentially toxic compounds, their modulation might have serious consequences. In this work, we studied selected antioxidant and drug-metabolizing enzymes (DME) in monosodium glutamate-mouse model of obesity. Specific activities, protein, and mRNA expressions of these enzymes in liver as well as in small intestine were compared in obese male mice and in their lean counterparts. Furthermore, expression of the NF-E2-related factor 2 (Nrf2) and its relation to obesity were tested. Obtained results showed that obesity affects expression and/or activities of some DME and antioxidant enzymes. In obese mice, upregulation of UDP-glucuronosyltransferases 1A (UGT1A), NAD(P)H:quinone oxidoreductase 1 (NQO1), nuclear transcription factor Nrf2, and downregulation of some isoforms of glutathione S-transferases (GST) were observed. Most of these changes were tissue and/or isoform specific. NQO1 seems to be regulated transcriptionally via Nrf2, but other enzymes might be regulated post-transcriptionally and/or post-translationally. Enhanced expression of Nrf2 in livers of obese mice is expected to play a role in protective adaptation. In contrast, elevated activities of NQO1 and UGT1A may cause alterations in drug pharmacokinetics in obese individuals. Moreover, decreased capacity of GST in obese animals indicates potentially reduced antioxidant defense and weaker chemoprotection.

• MeSH
• faktor 2 související s NF-E2 genetika   metabolismus   MeSH
• glukuronosyltransferasa genetika   metabolismus   MeSH
• glutamát sodný MeSH
• glutathiontransferasa genetika   metabolismus   MeSH
• inbrední kmeny myší MeSH
• izoenzymy genetika   metabolismus   MeSH
• játra enzymologie   metabolismus   MeSH
• messenger RNA metabolismus   MeSH
• modely nemocí na zvířatech * MeSH
• NAD(P)H dehydrogenasa (chinon) genetika   metabolismus   MeSH
• novorozená zvířata MeSH
• obezita enzymologie   metabolismus   MeSH
• orgánová specificita MeSH
• regulace genové exprese enzymů * MeSH
• střevní sliznice enzymologie   metabolismus   MeSH
• tenké střevo enzymologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Klíčová slova
• léčba DM založená na inkretinech,
• MeSH
• aminokyseliny, peptidy a proteiny fyziologie   metabolismus   sekrece   MeSH
• beta-buňky * enzymologie   metabolismus   sekrece   MeSH
• dipeptidylpeptidasa 4 farmakologie   metabolismus   sekrece   MeSH
• enteroendokrinní buňky cytologie   enzymologie   metabolismus   MeSH
• glukagonu podobný peptid 1 * fyziologie   metabolismus   sekrece   MeSH
• inkretiny fyziologie   izolace a purifikace   metabolismus   MeSH
• inzulin * fyziologie   izolace a purifikace   sekrece   MeSH
• L buňky (buněčná linie) fyziologie   metabolismus   účinky léků   MeSH
• Langerhansovy ostrůvky cytologie   fyziologie   sekrece   MeSH
• lidé MeSH
• lipidy fyziologie   sekrece   MeSH
• mastné kyseliny fyziologie   sekrece   MeSH
• nervový systém enzymologie   metabolismus   sekrece   MeSH
• potravní vláknina metabolismus   využití   MeSH
• sacharidy fyziologie   sekrece   MeSH
• statistika jako téma MeSH
• tenké střevo enzymologie   fyziologie   sekrece   MeSH
• žlučové kyseliny a soli fyziologie   metabolismus   sekrece   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

UNLABELLED: Consumption of antioxidant-enriched diets is 1 method of addressing obesity, which is associated with chronic oxidative stress and changes in the activity/expression of various enzymes. In this study, we hypothesized that the modulation of antioxidant enzymes and redox status through a cranberry extract (CBE)-enriched diet would differ between obese and nonobese mice. The CBE used in this study was obtained from the American cranberry (Vaccinium macrocarpon, Ericaceae), a popular constituent of dietary supplements that is a particularly rich source of (poly)phenols and has strong antioxidant properties. The present study was designed to test and compare the in vivo effects of 28-day consumption of a CBE-enriched diet (2%) on the antioxidant status of nonobese mice and mice with monosodium glutamate-induced obesity. Plasma, erythrocytes, liver, and small intestine were studied concurrently to obtain more complex information. The specific activities, protein, and messenger RNA expression levels of antioxidant enzymes as well as the levels of malondialdehyde and thiol (SH) groups were analyzed. Cranberry extract treatment increased the SH group content in plasma and the glutathione S-transferase activity in the erythrocytes of the obese and nonobese mice. In addition, in the obese animals, the CBE treatment reduced the malondialdehyde content in erythrocytes and increased NAD(P)H: quinone oxidoreductase (liver) and catalase (erythrocytes and small intestine) activities. The elevation of hepatic NAD(P)H: quinone oxidoreductase activity was accompanied by an increase in the corresponding messenger RNA levels. The effects of CBE on the activity of antioxidant enzymes and redox status were more pronounced in the obese mice compared with the nonobese mice.

• MeSH
• antioxidancia aplikace a dávkování   MeSH
• dieta MeSH
• erytrocyty chemie   MeSH
• glutathiontransferasa krev   MeSH
• játra enzymologie   MeSH
• katalasa krev   metabolismus   MeSH
• malondialdehyd krev   MeSH
• messenger RNA analýza   MeSH
• myši MeSH
• NAD(P)H dehydrogenasa (chinon) genetika   metabolismus   MeSH
• obezita krev   chemicky indukované   enzymologie   MeSH
• ovoce chemie   MeSH
• oxidace-redukce MeSH
• rostlinné extrakty aplikace a dávkování   MeSH
• sulfhydrylové sloučeniny krev   MeSH
• tenké střevo enzymologie   MeSH
• Vaccinium macrocarpon * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

OBJECTIVES: Dihydromyricetin (DHM) is a flavonoid, which has been shown to antagonize effects of ethanol intoxication. As a potential pharmacological agent, its biological interactions with enzymes metabolizing foreign compounds should be tested. Thus, the aim of this study was to analyze the influence of DHM on the induction and metabolic activity of selected cytochromes P450 (CYPs). METHODS: After flavonoid administration by oral gavage to stomach the CYP expression at protein and mRNA levels was determined in rat liver and small intestine. The effects of flavonoids on CYP1A1/2, CYP1A2 or CYP2B1/2 enzyme activities in microsomes were measured using marker activities of these enzymes. Flavonoid-mediated inhibition of recombinant CYP1A2 was also assayed with luciferin-ME substrate. The flavonoid interaction with aryl hydrocarbon receptor (AhR) was assayed by reporter luciferase activity in Hep2G cells. RESULTS: The value of half maximal inhibitory concentration of DHM for CYP1A1/2, CYP1A2, and CYP2B1 were determined to be 4.1, 14.2, and 98.5 mmol.L(-1), respectively. With the exception of a weak induction of CYP2B1 and CYP1A2 in the middle part of small intestine and in the liver, respectively, DHM did not affect the CYP expression at protein levels. On the contrary, real-time PCR revealed elevated expression of CYP1A1 and CYP1A2 mRNA in proximal part of the small intestine while decreased in the middle part. In the study utilizing the HepG2 cells, DHM showed only an additive effect on the benzo[a]pyrene-mediated activation of Ah receptor. CONCLUSIONS: Dihydromyricetin doesn't significantly interfere with metabolic activity of CYP1A1/2 and CYP2B1 enzymes.

• MeSH
• flavonoly farmakologie   MeSH
• inhibiční koncentrace 50 MeSH
• játra účinky léků   enzymologie   MeSH
• karcinogeny metabolismus   MeSH
• krysa rodu rattus MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• messenger RNA účinky léků   metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• potkani Wistar MeSH
• systém (enzymů) cytochromů P-450 účinky léků   genetika   metabolismus   MeSH
• tenké střevo účinky léků   enzymologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Dehydrogenase/reductase (SDR family) member 3 (DHRS3), also known as retinal short-chain dehydrogenase/reductase (retSDR1) is a member of SDR16C family. This family is thought to be NADP(H) dependent and to have multiple substrates; however, to date, only all-trans-retinal has been identified as a DHRS3 substrate. The reductive reaction catalysed by DHRS3 seems to be physiological, and recent studies proved the importance of DHRS3 for maintaining suitable retinoic acid levels during embryonic development in vivo. Although it seems that DHRS3 is an important protein, knowledge of the protein and its properties is quite limited, with the majority of information being more than 15 years old. This study aimed to generate a more comprehensive characterisation of the DHRS3 protein. Recombinant enzyme was prepared and demonstrated to be a microsomal, integral-membrane protein with the C-terminus oriented towards the cytosol, consistent with its preference of NADPH as a cofactor. It was determined that DHRS3 also participates in the metabolism of other endogenous compounds, such as androstenedione, estrone, and DL-glyceraldehyde, and in the biotransformation of xenobiotics (e.g., NNK and acetohexamide) in addition to all-trans-retinal. Purified and reconstituted enzyme was prepared for the first time and will be used for further studies. Expression of DHRS3 was shown at the level of both mRNA and protein in the human liver, testis and small intestine. This new information could open other areas of DHRS3 protein research.

• MeSH
• alkoholoxidoreduktasy metabolismus   MeSH
• cytosol metabolismus   MeSH
• jaterní mikrozomy enzymologie   metabolismus   MeSH
• játra enzymologie   metabolismus   MeSH
• lidé MeSH
• membránové proteiny metabolismus   MeSH
• NADH, NADPH oxidoreduktasy metabolismus   MeSH
• NADP metabolismus   MeSH
• Sf9 buňky MeSH
• Spodoptera metabolismus   MeSH
• syntázy mastných kyselin metabolismus   MeSH
• tenké střevo enzymologie   metabolismus   MeSH
• testis enzymologie   metabolismus   MeSH
• tretinoin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Matrix metalloproteinases (MMPs), responsible for extracellular matrix remodelling and processing of numerous soluble and cell-surface proteins, appear to play important roles in pathogenesis of gastrointestinal diseases. MMPs influence migration of inflammatory cells, mucosal destruction, matrix deposition and degradation. In this study, we analysed the expression of MMP-19 in the main forms of gastrointestinal diseases including inflammatory bowel diseases (IBD) such as ulcerative colitis and Crohn's disease, and colorectal carcinoma. We identified prominent MMP-19 expression in unaffected areas of intestinal epithelia and macrophages but not in other cells or tissues. Abundant expression of MMP-19 was also found in the endothelium of blood and lymphatic vessels of inflamed intestinal tissue. High MMP-19 immunoreactivity was also associated with macrophages in inflamed areas and myenteric plexuses. In comparison to the intestinal epithelium, all these cell types and compartments appeared to express MMP-19 irrespective of the disease pathogenesis and progression. Intestinal epithelia exhibited striking differential immunoreactivity for MMP-19. While immunoreactivity of monoclonal antibody recognizing the propeptide domain declined in virtually all IBD and colorectal carcinoma samples, other polyclonal antibodies against the hinge region and propetide domain did not show such an obvious decrease. Additional Western blotting analysis revealed that the antibodies against MMP-19 recognize differently processed forms of this MMP. The disappearance of immunoreactivity of the monoclonal anti-propeptide domain antibody does not mean down-regulation of MMP-19, but processing of the immature form. As this processing likely leads to the activation of this MMP, the differential staining pattern may be an important sign of disease progression.

• MeSH
• dospělí MeSH
• gastrointestinální nemoci enzymologie   patologie   MeSH
• HCT116 buňky MeSH
• kolon enzymologie   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• metaloproteinasy secernované do matrix metabolismus   MeSH
• mladý dospělý MeSH
• posttranslační úpravy proteinů * MeSH
• progrese nemoci * MeSH
• protilátky metabolismus   MeSH
• reprodukovatelnost výsledků MeSH
• senioři MeSH
• střevní sliznice enzymologie   patologie   MeSH
• tenké střevo enzymologie   patologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Benzo[a]pyrene (BaP) is a human carcinogen requiring metabolic activation prior to reaction with DNA. Cytochrome P450 (CYP) 1A1 is the most important hepatic and intestinal enzyme in both BaP activation and detoxification. CYP1A2 is also capable of oxidizing BaP, but to a lesser extent. The induction of CYP1A1/2 by BaP and/or β-naphthoflavone in liver and small intestine of rats was investigated. Both BaP and β-naphthoflavone induced CYP1A expression and increased enzyme activities in both organs. Moreover, the induction of CYP1A enzyme activities resulted in an increase in formation of BaP-DNA adducts detected by (32)P-postlabeling in rat liver and in the distal part of small intestine in vivo. The increases in CYP1A enzyme activity were also associated with bioactivation of BaP and elevated BaP-DNA adduct levels in ex vivo incubations of microsomes of both organs with DNA and BaP. These findings indicate a stimulating effect of both compounds on BaP-induced carcinogenesis.

• MeSH
• adukty DNA metabolismus   MeSH
• aktivace enzymů účinky léků   MeSH
• benzopyren toxicita   MeSH
• beta-naftoflavon farmakologie   MeSH
• chromatografie na tenké vrstvě MeSH
• cytochrom P-450 CYP1A1 metabolismus   MeSH
• cytochrom P-450 CYP1A2 biosyntéza   genetika   MeSH
• izoenzymy účinky léků   metabolismus   MeSH
• jaterní mikrozomy účinky léků   enzymologie   metabolismus   MeSH
• játra účinky léků   enzymologie   metabolismus   MeSH
• krysa rodu rattus MeSH
• látky znečišťující životní prostředí toxicita   MeSH
• potkani Wistar MeSH
• tenké střevo účinky léků   enzymologie   metabolismus   MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

OBJECTIVES: Benzo[a]pyrene (BaP) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) are carcinogens, which frequently occur in the human diet. Their metabolic activation to reactive species binding to DNA is mediated by cytochromes P450 (CYPs) 1A1 and 1A2. Thus, levels and activities of these CYPs are crucial for initiation of BaP- and PhPI-mediated carcinogenesis. Here, the effect of CYP1A1/2 induction due to their prototype flavonoid inducer, β-naphthoflavone (BNF), on BaP- and PhPI-derived DNA adduct formation in rats was examined. METHODS: Male rats pretreated with BNF were treated with a single dose of either carcinogen by oral gavage. Nuclease P1 version of 32P-postlabeling assay and online column-switching liquid chromatography-electrospray ionization-tandem mass spectrometry were used to detect and quantify covalent DNA adducts formed by BaP and PhIP in-vivo, respectively. Expression of CYP1A1/2 enzymes was examined by Western blot. Enzymatic activities of CYP1A1/2 were assessed using their marker substrates (ethoxyresorufin and methoxyresorufin). RESULTS: Treatment of rats with a single dose of BNF produced an increase in levels CYP1A1/2 and CYP1A1 proteins in liver and small intestine, respectively. An increase in CYP1A1 protein expression found in both organs correlated well with specific activities of these CYPs. The CYP1A1 expression levels and its specific activity in small intestine decreased along the length of the organ, being highest in its proximal part and lowest in its distal part. The BNF induction of CYP1A1/2 resulted in a significant increase in the formation of BaP- and PhIP-DNA adducts in liver and in the distal part of the small intestine, respectively. Thus, pretreatment of rats with BNF did not prevent the PhIP and BaP activation, but vice versa, enhanced their genotoxicity. CONCLUSIONS: The results of this study demonstrate that the administration of only a single dose of CYP-inducing flavonoid prior to the intake of food carcinogens may increase the risk of a tumor formation.

• MeSH
• beta-naftoflavon farmakologie   MeSH
• cytochrom P-450 CYP1A1 antagonisté a inhibitory   genetika   metabolismus   MeSH
• enzymová indukce účinky léků   MeSH
• inhibitory enzymů farmakologie   MeSH
• játra účinky léků   enzymologie   metabolismus   MeSH
• karcinogeny životního prostředí izolace a purifikace   toxicita   MeSH
• kontaminace potravin analýza   MeSH
• krysa rodu rattus MeSH
• poškození DNA účinky léků   MeSH
• potkani Wistar MeSH
• potraviny toxicita   MeSH
• preklinické hodnocení léčiv MeSH
• tenké střevo účinky léků   enzymologie   metabolismus   MeSH
• testy genotoxicity MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH