Vaccinium myrtillus L. (bilberry) fruit is a blue-colored berry with a high content of anthocyanins. These bioactive secondary metabolites are considered to play a major role in the health-promoting properties of bilberries. Our in vivo study was designed to assess the possible influence of bilberry extract on drug-metabolizing enzymes (DMEs). Rats were exposed to bilberry extract in drinking water at two concentrations (0.15 and 1.5 g/L). Selected DMEs were determined (mRNA expression and enzymatic activity) after 29 and 58 days in rat liver. In addition, a panel of antioxidant, physiological, biochemical and hematological parameters was studied; these parameters did not demonstrate any impact of bilberry extract on the health status of rats. A significant increase in activity was observed in cytochrome P450 (CYP) 2C11 (131% of control) and CYP2E1 (122% of control) after a 29-day administration, while the consumption of a higher concentration for a longer time led to a mild activity decrease. Slight changes were observed in some other DMEs, but they remained insignificant from a physiological perspective. According to our results, we conclude that the consumption of bilberries as a food supplement should not pose a risk of interacting with co-administered drugs based on their metabolism.
- MeSH
- antioxidancia farmakologie MeSH
- krysa rodu rattus MeSH
- potkani Wistar MeSH
- rostlinné extrakty farmakologie MeSH
- systém (enzymů) cytochromů P-450 účinky léků metabolismus MeSH
- Vaccinium myrtillus chemie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The prevalence of obesity and other obesity-related diseases is increasing worldwide. Obesity is a disease characterized by increased body weight, or a condition resulting from excessive accumulation of body fat. Due to increased body fat deposits, obesity has also been associated with increased mortality resulting from higher incidence rates of hypertension, diabetes, or various types of cancer, such as breast, colorectal, cervical and prostate cancer. Physiological changes associated with obesity are likely to result in altered drug biotransformation. The main enzymes enabling the oxidative biotransformation of most drugs are cytochromes P450 (CYPs). The review summarizes how pathophysiological factors, especially obesity, affect properties (e.g. enzyme activity, protein expression, gene expression) of CYP enzymes in various experimental models of human obesity. Results reported by various authors suggest that obesity is associated with a decrease of CYP activities (except for the CYP2C and CYP2E1 enzymes). The only exception is mouse obesity induced by monosodium glutamate (administered to newborn mice) as it usually leads to increased CYP expression. Selecting an animal model that is as close as possible to the properties of human obesity is of paramount importance.
The influence of metabolites of sulforaphane, natural compounds present in broccoli (Brassica oleracea var. botrytis italica) and in other cruciferous vegetables, on drug-metabolizing cytochrome P450 (CYP) enzymes in human liver microsomes and possible entry of sulforaphane into human hepatic cells were investigated. Metabolites studied are compounds derived from sulforaphane by the mercapturic acid pathway (conjugation with glutathione and by following reactions), namely sulforaphane glutathione and sulforaphane cysteine conjugates and sulforaphane-N-acetylcysteine. Their possible effect on four drug-metabolizing CYP enzymes, CYP3A4 (midazolam 1'-hydroxylation), CYP2D6 (bufuralol 1'-hydroxylation), CYP1A2 (7-ethoxyresorufin O-deethylation), and CYP2B6 (7-ethoxy-4-(trifluoromethyl)coumarin O-deethylation), was tested. Inhibition of four prototypical CYP activities by sulforaphane metabolites was studied in pooled human liver microsomes. Sulforaphane metabolites did not considerably affect biological function of drug-metabolizing CYPs in human liver microsomes except for CYP2D6, which was found to be inhibited down to 73-78% of the original activity. Analysis of the entry of sulforaphane into human hepatocytes was done by cell disruption by sonication, methylene chloride extraction, and modified high-performance liquid chromatography method. The results have shown penetration of sulforaphane into the human hepatic cells.
- MeSH
- Brassica chemie MeSH
- cystein metabolismus MeSH
- cytochrom P-450 CYP2D6 účinky léků metabolismus MeSH
- glutathion metabolismus MeSH
- hepatocyty metabolismus MeSH
- inhibitory cytochromu P450 farmakologie MeSH
- isothiokyanatany analýza metabolismus farmakologie MeSH
- jaterní mikrozomy enzymologie MeSH
- játra chemie metabolismus MeSH
- lidé MeSH
- systém (enzymů) cytochromů P-450 účinky léků metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Účel studie: Solidago virgaurea „zlatobýl“ je léčivá rostlina zajímavá z pohledu biologických účinků, známých již ve starověku (1, 2). Obsahovými látkami rodu Solidago jsou flavonoidy, fenolové kyseliny a jejich glykosidy, polysacharidy a další sloučeniny (3). Je známo, že Solidago virgaurea vykazuje diuretické, protizánětlivé a nově popsané protinádorové, antimikrobiální, sedativní a hypotenzní účinky (1, 2). Zejména flavonoidy a fenolický diglukosid – leiokarposid patří mezi látky, u kterých byla studová- na řada pozitivních účinků na organizmus; u leiokarposidu je znám mimo jiné jeho vliv na diurézu (3). Cílem této studie bylo zjistit, zda extrakt, odvar a výluh z nati zlatobýlu obecného, Solidaginis virgaureae herba, ovlivňují vlastnosti enzymů I. fáze biotransformace, zejména cytochromů P450 v lidské jaterní mikrozomální frakci a v primárních kulturách lidských hepatocytů. Použité metody: Byly studovány různé koncentrace extraktu, odvaru a výluhu ze Solidaginis virgaureae herba, vyjádřené v mg sušiny na ml vody (0,042 mg/ml; 0,083 mg/ml a 0,167 mg/ml). Enzymové aktivity byly stanoveny pomocí metody HPLC; exprese proteinů pomocí metody Western blotu. Výsledky: Aktivity a exprese proteinů CYP1A2, CYP2A6, CYP2C9, CYP2D6 a CYP3A4 vykazovaly relativně malé změny po ovlivnění testovanými vzorky v lidských hepatocytech i v lidské jaterní mikrozomální frakci, ve všech studovaných koncentracích. Závěr: Na základě získaných výsledků enzymových aktivit a množství proteinu lze předpokládat, že testované vzorky signifikantně neovlivňují metabolizmus současně podávaných léčiv a jejich užívání pravděpodobně nevede ke klinicky významným lékovým interakcím.
Purpose: Solidago virgaurea „goldenrod“ is a known medicinal plant that has been used since ancient times (1, 2). The genus Solidago is interesting in terms of both biological effects and the presence of many interesting secondary metabolites – flavonoids, phenolic acids and glucosides, polysaccharides and others (3). Solidago virgaurea is known to exhibit diuretic, antiinflammatory and newly described antitumoral, antimicrobial, sedative and hypotensive effects (1, 2). Primarily flavonoids and phenolic diglucoside – leiocarposide belong to secondary metabolites having primarily a positive effect on diuresis (3). Our objective was to investigate whether Solidaginis virgaureae herba extract, decoction and leachate affect the properties of phase I biotransformation enzymes, namely cytochromes P450 in human liver microsomal fraction and also in primary cultures of human hepatocytes. Methods: Different concentrations of Solidaginis virgaureae herba extract, decoction and leachate, expressed as mg of dry weight in mL of water (0.042 mg/mL; 0.083 mg/mL and 0.167 mg/mL) were studied. Enzyme activities and protein expression were determined using HPLC and Western blotting, respectively. Results: Activity and protein expression of CYP1A2, CYP2A6, CYP2C9, CYP2D6 and CYP3A4 were only little influenced by Solidaginis virgaureae herba preparations, either in human hepatocytes as well as in human liver microsomal fraction in all studied concentrations. Conclusion: Based on the results of the enzyme activities and the amount of protein can be expected that the Solidaginis virgaureae herba preparations do not significantly affect the metabolism of concomitantly administered drugs and their use probably does not result in drug interactions.
- MeSH
- biotransformace * MeSH
- enzymatické testy MeSH
- fytoterapie MeSH
- interakce bylin a léků MeSH
- jaterní mikrozomy enzymologie účinky léků MeSH
- léčivé rostliny MeSH
- lidé MeSH
- rostlinné extrakty * MeSH
- Solidago * MeSH
- stonky rostlin MeSH
- subcelulární frakce enzymologie účinky léků MeSH
- systém (enzymů) cytochromů P-450 účinky léků MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- western blotting MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
PURPOSE: Consumption of dietary supplements with green tea extract (GTE) is popular for weight management, but it may be accompanied by various side effects, including interactions with drugs. The aim of the present in vivo study was to evaluate the effect of defined GTE (Polyphenon 60) in three dosage schemes on insulin, leptin and drug-metabolizing enzymes in obese mice. METHODS: Experimental obesity was induced by repeated s.c. application of monosodium glutamate to newborn mice. Green tea extract was administered in three dosage schemes in chow diet. The plasmatic levels of insulin and leptin were assayed using enzyme-linked immunosorbent assay. Enzyme activities and mRNA expressions of drug-metabolizing enzymes (totally 13) were analyzed in liver and small intestine using spectrophotometric and HPLC assays and RT-PCR, respectively. RESULTS: GTE-treatment decreased insulin and leptin levels. Eleven enzymes were significantly affected by GTE-treatment. Long-term administration of 0.01% GTE caused increase in the activity and mRNA level of cytochrome P450 3A4 (CYP3A4) ortholog in the liver as well as in the small intestine. Interestingly, short-term overdose by GTE (0.1%) had more pronounced effects on enzyme activities and mRNA expressions than long-term overdose. CONCLUSIONS: GTE-mediated induction of CYP3A4 ortholog, the main drug-metabolizing enzyme, could result in decreased efficacy of simultaneously or subsequently administered drug in obese individuals.
- MeSH
- antioxidancia farmakologie MeSH
- aromatické hydroxylasy genetika metabolismus MeSH
- čaj chemie MeSH
- cytochrom P-450 CYP2E1 genetika metabolismus MeSH
- cytochrom P-450 CYP3A genetika metabolismus MeSH
- ELISA MeSH
- glutamát sodný škodlivé účinky MeSH
- inzulin krev MeSH
- játra účinky léků metabolismus MeSH
- leptin krev MeSH
- messenger RNA genetika metabolismus MeSH
- modely nemocí na zvířatech MeSH
- myši obézní MeSH
- myši MeSH
- obezita chemicky indukované farmakoterapie MeSH
- potravní doplňky * MeSH
- rostlinné extrakty farmakologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cytochrome P450 (CYP) expression and activity are not homogeneous in the liver lobules. Indeed, CYPs are mainly expressed and induced in centrilobular hepatocytes. The wingless-type MMTV integration site family (WNT)/β-catenin pathway was identified as a major regulator of this zonal organization. We have recently demonstrated that in primary human hepatocytes (PHHs), the expression of CYP2E1, CYP1A2, and aryl hydrocarbon receptor (AhR), but not of CYP3A4, is regulated by the WNT/β-catenin pathway in response to WNT3a, its canonical activator. Here, we investigated whether glycogen synthase kinase 3β (GSK3β) inhibitors, which mimic the action of WNT molecules, could be used in PHHs to activate the β-catenin pathway to study CYP expression. We assessed the activity of 6BIO (6-bromoindirubin-3'-oxime), CHIR99021 (6-((2-((4-(2,4-dichlorophenyl)-5-(4methyl-1H-imidazol-2-yl)pyrimidin-2-yl)amino)ethyl)amino) nicotinonitrile), and GSK3iXV (Pyridocarbazolo-cyclopentadienyl Ruthenium complex GSK3 inhibitor XV) that belong to structurally different families of GSK3β inhibitors. Using small interfering RNAs, reporter gene assays, and molecular docking predictions, we demonstrated that GSK3β inhibitors can activate the WNT/β-catenin pathway in PHHs to regulate CYP2E1 expression. We also found that 6BIO and GSK3iXV are AhR full agonists that participate, through AhR signaling, to CYP1A2 induction. Conversely, CHIR99021 is an AhR partial agonist, and a pregnane X receptor ligand and partial agonist, thus regulating CYP1A2 and CYP3A4 gene expression in a β-catenin-independent manner. In conclusion, GSK3β inhibitors can activate the WNT/β-catenin pathway in PHHs. Nevertheless, their role in CYP regulation should be analyzed with caution as these molecules can interact with xenosensors.
- MeSH
- beta-katenin agonisté antagonisté a inhibitory genetika metabolismus MeSH
- enzymová indukce účinky léků MeSH
- hepatocyty cytologie účinky léků metabolismus MeSH
- indoly farmakologie MeSH
- induktory cytochromu P450 chemie metabolismus farmakologie MeSH
- inhibitory proteinkinas chemie metabolismus farmakologie MeSH
- kinasa 3 glykogensynthasy antagonisté a inhibitory metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- organokovové sloučeniny farmakologie MeSH
- oximy farmakologie MeSH
- pyridiny farmakologie MeSH
- pyrimidiny farmakologie MeSH
- receptory aromatických uhlovodíků agonisté chemie genetika metabolismus MeSH
- rekombinantní fúzní proteiny chemie metabolismus MeSH
- reportérové geny účinky léků MeSH
- RNA interference MeSH
- signální dráha Wnt účinky léků MeSH
- simulace molekulového dockingu MeSH
- steroidní receptory agonisté genetika metabolismus MeSH
- systém (enzymů) cytochromů P-450 chemie genetika metabolismus MeSH
- transkripční faktory bHLH agonisté chemie genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
UNLABELLED: Cytochromes P450 (CYPs) are enzymes present from bacteria to man involved in metabolism of endogenous and exogenous compounds incl. drugs. Our objective was to assess whether obesity leads to changes in activities and expression of CYPs in the mouse liver, small intestine and colon. MAIN METHODS: An obese mouse model with repeated injection of monosodium glutamate (MSG) to newborns was used. Controls were treated with saline. All mice were sacrificed at 8 months. In the liver and intestines, levels of CYP mRNA and proteins were analyzed using RT-PCR and Western blotting. Activities of CYP enzymes were measured with specific substrates of human orthologous forms. KEY FINDINGS: At the end of the experiment, body weight, plasma insulin and leptin levels as well as the specific content of hepatic CYP enzymes were increased in obese mice. Among CYP enzymes, hepatic CYP2A5 activity, protein and mRNA expression increased most significantly in obese animals. Higher activities and protein levels of hepatic CYP2E1 and 3A in the obese mice were also found. No or a weak effect on CYPs 2C and 2D was observed. In the small intestine and colon, no changes of CYP enzymes were detected except for increased expression of CYP2E1 and decreased expression of CYP3A mRNAs in the colon of the obese mice. SIGNIFICANCE: Results of our study suggest that the specific content and activities of some liver CYP enzymes (especially CYP2A5) can be increased in obese mice. Higher activity of CYP2A5 (CYP2A6 human ortholog) could lead to altered metabolism of drug substrates of this enzyme (valproic acid, nicotine, methoxyflurane).
- MeSH
- glutamát sodný * MeSH
- inzulin krev metabolismus MeSH
- játra enzymologie metabolismus MeSH
- krevní glukóza metabolismus MeSH
- leptin krev metabolismus MeSH
- lidé MeSH
- lipidy krev MeSH
- messenger RNA analýza genetika MeSH
- metabolismus lipidů MeSH
- myši MeSH
- obezita krev chemicky indukované genetika metabolismus MeSH
- přijímání potravy MeSH
- střeva enzymologie metabolismus MeSH
- systém (enzymů) cytochromů P-450 analýza genetika metabolismus MeSH
- upregulace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
