Papers by Zuzana Matušková
The exposure of human cells to oxidative stress leads to the oxidation of biomolecules such as li... more The exposure of human cells to oxidative stress leads to the oxidation of biomolecules such as lipids, proteins and nuclei acids. In this study, the oxidation of lipids, proteins and DNA was studied after the addition of hydrogen peroxide and Fenton reagent to cell suspension containing human leukemic monocyte lymphoma cell line U937. EPR spin-trapping data showed that the addition of hydrogen peroxide to the cell suspension formed hydroxyl radi-cal via Fenton reaction mediated by endogenous metals. The malondialdehyde HPLC anal-ysis showed no lipid peroxidation after the addition of hydrogen peroxide, whereas the Fenton reagent caused significant lipid peroxidation. The formation of protein carbonyls monitored by dot blot immunoassay and the DNA fragmentation measured by comet assay occurred after the addition of both hydrogen peroxide and Fenton reagent. Oxidative dam-age of biomolecules leads to the formation of singlet oxygen as conformed by EPR spin-trapping spectroscopy and th...
Cilem předkladane disertacni prace bylo zjistit, zda probioticke bakterie, Escherichia coli Nissl... more Cilem předkladane disertacni prace bylo zjistit, zda probioticke bakterie, Escherichia coli Nissle 1917 a Lactobacillus casei, ovlivňuji cytochromy P450 (zkraceně CYP, nejdůležitějsi enzymy metabolizmu cizorodých latek vcetně leciv) v jatrech a ve střevě potkana. V prvni casti prace byla sledovana enzymova aktivita, exprese mRNA a exprese proteinů jednotlivých forem CYP (k tomu byla rovněž provedena studie distribuce CYP v potkanim střevě) po premedikaci oběma probiotiky. Ve druhe casti prace bylo studovano, jestli aplikovana probiotika mohou alterovat farmakokineticke parametry modelových leciv (amiodaronu a diklofenaku). V předběžne studii distribuce forem CYP v potkanim střevě bylo zjistěno, že se zastoupeni jednotlivých vybraných enzymů CYP po cele delce střevniho traktu potkana měni. V tenkem střevě se nachazi předevsim CYP1A1 a 2B1/2, zatimco CYP2E1 je ve vysokých koncentracich jak v tenkem střevě, tak i ve slepem. Naopak CYP2C6 se nachazi předevsim v tlustem střevě (caecum i ...
PLOS ONE, 2015
<p>Data are presented as mean values and standard deviations. The mean value represents the... more <p>Data are presented as mean values and standard deviations. The mean value represents the average value from at least three measurements.</p><p>Analysis of the medians of comet, head, and tail parameters in the control, the H<sub>2</sub>O<sub>2</sub>-treated and the Fenton reagent-treated U937 cells.</p
<p>The cell viability was determined 30 min after the addition of 5 mM H<sub>2</su... more <p>The cell viability was determined 30 min after the addition of 5 mM H<sub>2</sub>O<sub>2</sub> or Fenton reagent to the U937 cells. The results are normalized to control U937 cells. The data are presented as the mean and standard deviation of at least 3 measurements.</p
Klinická farmakologie a farmacie, 2018
Prezentovaná studie měla za cíl porovnat obsah silybinu ve vybraných potravních doplňcích obsahuj... more Prezentovaná studie měla za cíl porovnat obsah silybinu ve vybraných potravních doplňcích obsahujících silymarin dostupných v České republice. Stanovení silybinu bylo provedeno pomocí HPLC/UV. Naše výsledky ukazují, že většina potravních doplňků obsahuje silybin v referenčním rozmezí 20-40 %. Výrobky NaturProdukt® Silymarin forte s obsahem 40 % silybinu (80,00 ± 0,50 mg) a Nefdesanté® Silymarin ostropestřec s obsahem 33 % silybinu (81,58 ± 0,14 mg) patří mezi výrobky s vysokým obsahem této přírodní látky. Na druhou stranu nejméně silybinu bylo naměřeno u přípravku WALMARK® Ostropestřec mariánský (< 20 %). Výsledky naznačují, že většina z testovaných potravních doplňků je kvalitní a obsahuje deklarované množství obsahových látek. Klíčová slova: silymarin, silybin, potravní doplňky. Determination of silybin content in food supplements containing silymarin The aim of this study was to determine the amount of silybin in selected nutraceuticals containing silymarin available in the Czech Republic. The determination of silybin was performed using HPLC/UV. Results show that almost all of the tested nutraceuticals contain the sufficient amount of silybin (range 20-40 %). NaturProdukt® Silymarin forte contained 40 % of silybin (80.00 ± 0.50 mg) and Nefdesanté® Silymarin ostropestřec contained 33 % of silybin (81.58 ± 0.14 mg). Both can be included among the products with high silybin content. On the other hand, the lowest amount of silybin was measured in WALMARK® Ostropestřec mariánský (< 20 %). Our results suggest that almost all of the tested products are of high quality and contain the declared amount of constituents.
<p>The SOSG fluorescence from U937 cells was measured after the addition of 5 mM H<sub&g... more <p>The SOSG fluorescence from U937 cells was measured after the addition of 5 mM H<sub>2</sub>O<sub>2</sub> and Fenton reagent (5 mM H<sub>2</sub>O<sub>2</sub> and 1 mM FeSO<sub>4</sub>) by confocal laser scanning microscopy. 50 μM SOSG was added 30 min prior to the data collection to the U937 cells. In A, left column shows the combination of Nomarski DIC and SOSG fluoresencence (λ<sub>em</sub> = 505–525 nm) channels and right column the integral distribution of SOSG fluorescence. In B, the intensity of SOSG fluorescence (upper graph) and the percentage of cells showing the SOSG fluorescence (lower graph) are shown. Data are presented as mean values and standard deviations. The mean value represents the average value from five to ten representative microphotographs per variant.</p
<p>Dot blot membrane in the control (A), the H<sub>2</sub>O<sub>2</sub... more <p>Dot blot membrane in the control (A), the H<sub>2</sub>O<sub>2</sub>-treated (B) and the Fenton reagent-treated (C) U937 cells. The U937 cells were treated with 5 mM H<sub>2</sub>O<sub>2</sub> (B) and Fenton reagent (5 mM H<sub>2</sub>O<sub>2</sub> and 1 mM FeSO<sub>4</sub>) (C) for 30 min. After the treatment, the supernatant containing proteins were pipetted on the membrane and incubated with primary antibody (anti-dinitrophenyl-KLH, rabbit igG fraction, biotinilated) overnight. Furthermore the membrane was incubated with streptavidin-HRP conjugate for 1 h, developed with ECL and chemiluminescence signal was detected on X-ray film. The densitometry results are 0.5±0.2 (control), 1.4±0.2 (H<sub>2</sub>O<sub>2</sub>-treated), and 2.3±0.3 (Fenton reagent). Data are presented as mean values and standard deviations. The mean value represents the average value from at least three measurements.</p
<p>The chromatogram of DNPH-MDA complex in U937 cells (A) and DNPH-MDA standard (B). In A, ... more <p>The chromatogram of DNPH-MDA complex in U937 cells (A) and DNPH-MDA standard (B). In A, chromatogram of DNPH-MDA complex was measured in the control (trace a), the H<sub>2</sub>O<sub>2</sub>-treated (trace b) and the Fenton reagent-treated (trace c) U937 cells. The U937 cells were treated with 5 mM H<sub>2</sub>O<sub>2</sub> (b) and Fenton reagent (5 mM H<sub>2</sub>O<sub>2</sub> and 1 mM FeSO<sub>4</sub>) (c) for 30 min. After the treatment, lipids were separated from proteins and DNPH was added to lipids. In B, the chromatogram of DNPH-MDA standard shows the retention time of 3 min 50 s. The insert shows the dependence of average peak area on the concentration of DNPH-MDA standard. Based on the calibration curve, the concentrations of DNPH-MDA complex determined from calibration curve were as following: 0.029±0.003 nmol ml<sup>−1</sup> (control), 0.030±0.003 (H<sub>2</sub>O<sub>2</sub>) and 0.09±0.02 nmol ml<sup>−1</sup> (Fenton reagent). The coefficient of determination R<sup>2</sup> was determined as 0.9997. Data are presented as mean values and standard deviations. The mean value represents the average value from at least three measurements.</p
<p>TEMPONE EPR spectra were measured in the control (A and B), the H<sub>2</sub>... more <p>TEMPONE EPR spectra were measured in the control (A and B), the H<sub>2</sub>O<sub>2</sub>-treated (C and D) and the Fenton reagent-treated (E and F) U937 cells in the presence of 100 mM TEMPD. U937 cells were treated with no addition (A), 5 mM H<sub>2</sub>O<sub>2</sub> (C) and Fenton reagent (5 mM H<sub>2</sub>O<sub>2</sub> and 1 mM FeSO<sub>4</sub>) (E) for a period indicated in Fig. In C and E, top traces show the simulation of TEMPONE EPR signal using hyperfine coupling constants <i>a</i><sup>N</sup> = 16 G. Chemical source of <sup>1</sup>O<sub>2</sub> (10 mM molybdic acid + 10 mM H<sub>2</sub>O<sub>2</sub> measured right after the preparation) was used as a positive control (C,E). Bar graphs represent the hight of the middle peak of TEMPONE EPR signal in the control (B), the H<sub>2</sub>O<sub>2</sub>-treated (D) and the Fenton reagent-treated (F) U937 cells. Experimental EPR conditions were as follows: microwave power, 10 mW; modulation amplitude, 1 G; modulation frequency, 100 kHz; sweep width, 100 G; scan rate, 1.62 G s-1, gain 500. Bars represent 4000 (A) and 8000 (C and E) relative units. Data are presented as mean values and standard deviations. The mean value represent the average value from at least three measurements.</p
Silymarin is a standardized extract from Milk Thistle (Silybum marianum). This extract is used in... more Silymarin is a standardized extract from Milk Thistle (Silybum marianum). This extract is used in supportive therapy of liver diseases for centuries (Rainone 2005) and showed positive effects on lipoprotein profile (Škottová et al 2003). This work studied effect of silymarin on rat cytochrome P450 - CYP2C6. This cytochrome P450 can also be regarded as a counterpart of human CYP2C9 (Daniel et al 2006). Rat CYP2C6 and human CYP2C9 are known to share some substrates (both catalyze the 4 ́-hydroxylation of diclofenac (Leemann et al 1993) and 7-hydroxylation of warfarin (Daniel et al 2006)). High significance of human CYP2C9 in metabolism of many drugs (ibuprofen, diclofenac, warfarin and others) is well documented (Mo et al 2009). This work aims to investigate effect of silymarin (1 % w/w), administered in high cholesterol and high fat diet (HCD) for 21 days, on the expression of liver CYP2C6 in hereditary hypertriglyceridemic (HHTg) rats. These animals selected from Wistar rats exhibit...
Silymarin is a standardised extract from Milk Thistle (Silybum marianum). This extract has been u... more Silymarin is a standardised extract from Milk Thistle (Silybum marianum). This extract has been used in supportive therapy of liver diseases for centuries (Rainone 2005) and shows positive effects on lipoprotein profile (Škottová et al. 2003). Our study studied effect of silymarin on rat cytochrome P450 CYP2E1. This cytochrome P450 metabolizes many substances to more toxic metabolites. For example paracetamol is converted to a highly toxic metabolite, N-acetyl-p-benzoquinone imine (NAPQI). This study aims to investigate the effect of silymarin (standardised silymarin, micronised silymarin and Phytosome silymarin (1 % w/w)), administered in standard laboratory diet (STD) for 4 weeks, on the expression of liver CYP2E1 in a hereditary hypertriglyceridemic (HHTg) rats. These animals breed from Wistar rats, exhibit hypertriglyceridemia, insulin resistance, hypertension and liver steatosis. This strain of rat represents an accepted model of metabolic syndrome (Vrána, Kazdová 1990). Our re...
Mezi významné faktory metabolického syndromu se řadí poruchy metabolismu lipidů, které jsou vysoc... more Mezi významné faktory metabolického syndromu se řadí poruchy metabolismu lipidů, které jsou vysoce asociovány s kardiovaskulárním rizikem. Jednou z možných léčebných metod jsou kromě léčiv s hypolipidemickým působením i látky přírodního původu. V řadě studií bylo potvrzeno pozitivní působení rostlinných extraktů s obsahem polyfenolů. Na polyfenolické látky a flavonoidy je bohatý bodlák z čeledi Asteraceae, Silybum marianum, a především pak extrakt z jeho semen - silymarin. V naší studii jsme porovnávali rozdílnou účinnost jednotlivých lékových forem silymarinu (standardizovaný extrakt silymarinu, mikronizovaný silymarin a silymarin ve formě phytosomů) v ovlivnění plazmatických hladin triacylglycerolů, celkového cholesterolu a HDL cholesterolu u hereditárně hypertriglyceridemických (HHTg) potkanů (akceptovaný model metabolického syndromu). U všech skupin potkanů krmených standardní dietou s obsahem různých lékových forem silymarinu došlo ke statisticky významnému poklesu plazmatickýc...
Mezi významné faktory metabolického syndromu se řadí poruchy metabolismu lipidů, které jsou vysoc... more Mezi významné faktory metabolického syndromu se řadí poruchy metabolismu lipidů, které jsou vysoce asociovány s kardiovaskulárním rizikem. Jednou z možných léčebných metod jsou kromě léčiv s hypolipidemickým působením i látky přírodního původu. V řadě studií bylo potvrzeno pozitivní působení rostlinných extraktů s obsahem polyfenolů. Na polyfenolické látky a flavonoidy je bohatý bodlák z čeledi Asteraceae, Silybum marianum, a především pak extrakt z jeho semen - silymarin. V naší studii jsme porovnávali rozdílnou účinnost jednotlivých lékových forem silymarinu (standardizovaný extrakt silymarinu, mikronizovaný silymarin a silymarin ve formě phytosomů) v ovlivnění plazmatických hladin triacylglycerolů, celkového cholesterolu a HDL cholesterolu u hereditárně hypertriglyceridemických (HHTg) potkanů (akceptovaný model metabolického syndromu). U všech skupin potkanů krmených standardní dietou s obsahem různých lékových forem silymarinu došlo ke statisticky významnému poklesu plazmatickýc...
Rosuvastatin is one of the latest statins (HMG-CoA inhibitors) used in cholesterol-lowering thera... more Rosuvastatin is one of the latest statins (HMG-CoA inhibitors) used in cholesterol-lowering therapy. Several statins are metabolized by cytochrome P450 (CYP) 3A4 and can therefore interact with commonly used medications. Rosuvastatin undergoes only minor metabolism by CYP enzymes (CYP2C9 and also CYP2C19) in man. 1 The aim of the study was to investigate whether rosuvastatin at a dose 0.01% affects rat CYP2C11 and CYP2C6 (counterpart of human CYP2C9). 2, 3 Male Wistar rats were kept for 3 weeks (ad libitum) on a standard laboratory diet (STD) or on an experimental high-cholesterol diet (HCD) prepared by adding 1% (w/w) of cholesterol and 10% (w/w) of lard fat to the STD. Rosuvastatin ( 0.01%, w/w) was administered as a dietary supplement to the HCD. Rats were then fasted overnight, i. m. anesthetized and their liver was removed and frozen. Relative amount of CYP2C11 and CYP2C6 protein in the liver microsomes was determined by Western blotting, hydroxylating activities of these enzym...
Microorganisms Escherichia coli Nissle 1917 (serotype O6:K5:H1) as a probiotic have beneficial ef... more Microorganisms Escherichia coli Nissle 1917 (serotype O6:K5:H1) as a probiotic have beneficial effects on the gastrointestinal tract [1]. The aim of this study was to find whether E. coli Nissle 1917 influences pharmacokinetics of concomitantly taken pharmacotherapeutic agent. An antiarrhythmic drug amiodarone was used in this study. The probiotic strain E. coli Nissle 1917 was administered to thirty male Wistar rats for seven days and the saline solution was administered to another thirty experimental animals for seven days (as a control group) as well. The eighth day, the amiodarone hydrochloride was applied to all sixty rats. Then, after 1, 2, 3, 4, 5.5, 7, 9, 14, 22 and 30 hours, the blood samples were taken from rat abdominal aorta; afterwards, the plasma level of amiodarone and of its metabolite N-desethylamiodarone was measured by HPLC-UV. The results show that the plasma level of amiodarone is higher after application of E. coli Nissle 1917 to rats. Apparently, the applicati...
Chybné ISSN 1211-166X patří časopisu: "Zpravodaj klinické farmakologie a farmacie" Podn... more Chybné ISSN 1211-166X patří časopisu: "Zpravodaj klinické farmakologie a farmacie" Podnázev do r. 2003: odborný časopis Sekce klinické farmakologie České společnosti experimentální a klinické farmakologie a toxikologie JEP Kazuistiky, zprávy ze sjezdů, varia určené nejen farmakologům. Součástí článků anglické abstrakty
Xenobiotica, 2015
1. To compare the effectiveness of different drug forms of silymarin: standardized extract of sil... more 1. To compare the effectiveness of different drug forms of silymarin: standardized extract of silymarin (SS), micronized silymarin (MS) and silymarin in the form of phytosome (PS) on dyslipidemia and liver fat accumulation in a model of metabolic syndrome, in non-obese hereditary hypertriglyceridemic rats. The second aim of this study was to slightly uncover the silymarin action on enzymes and proteins involved in cholesterol metabolism and excretion. 2. Silymarin administered to hereditary hypertriglyceridemic rats as dietary supplements (1%) for 4 weeks significantly lowered the plasma levels of triglycerides, total cholesterol and markedly increased HDL cholesterol level. Western blot analyses showed significant increase in the protein expression of CYP7A1 and CYP4A and increase in protein expression of selected ABC transporters. Silymarin in the form of phytosome and micronized silymarin were more effective forms of silymarin. 3. These findings suggest that silymarin may favorably affect the metabolism of cholesterol and triglycerides in rats with metabolic syndrome. Raising HDL levels suggests potentially important anti-atherogenic effect of silymarin. The changes in expression of cytochromes P450 and ABC transporters involved in cholesterol metabolism and excretion could be partially responsible for the hypolipidemic effect of silymarin.
PLOS ONE, 2015
The exposure of human cells to oxidative stress leads to the oxidation of biomolecules such as li... more The exposure of human cells to oxidative stress leads to the oxidation of biomolecules such as lipids, proteins and nuclei acids. In this study, the oxidation of lipids, proteins and DNA was studied after the addition of hydrogen peroxide and Fenton reagent to cell suspension containing human leukemic monocyte lymphoma cell line U937. EPR spin-trapping data showed that the addition of hydrogen peroxide to the cell suspension formed hydroxyl radical via Fenton reaction mediated by endogenous metals. The malondialdehyde HPLC analysis showed no lipid peroxidation after the addition of hydrogen peroxide, whereas the Fenton reagent caused significant lipid peroxidation. The formation of protein carbonyls monitored by dot blot immunoassay and the DNA fragmentation measured by comet assay occurred after the addition of both hydrogen peroxide and Fenton reagent. Oxidative damage of biomolecules leads to the formation of singlet oxygen as conformed by EPR spintrapping spectroscopy and the green fluorescence of singlet oxygen sensor green detected by confocal laser scanning microscopy. It is proposed here that singlet oxygen is formed by the decomposition of high-energy intermediates such as dioxetane or tetroxide formed by oxidative damage of biomolecules.
Pig liver microsomal cytochromes P450 seem to be at least in several cases good models of their h... more Pig liver microsomal cytochromes P450 seem to be at least in several cases good models of their human orthologs. Namely, the pig CYP3A29 mimic rather closely the properties of the human CYP3A4 form, pig CYP2A19 this of the human CYP2A6, and the pig and human CYP2E1 forms are also rather close in their function. This is probably not true for the CYP2C and CYP2D enzymes. Model substrates for the newly isolated pig CYP forms 2A19 and 1A revealed also similarity in the metabolism by CYP forms of the man and the pig. Skatole is a relatively important substrate of pig CYP enzymes as it contributes to the boar taint. Skatole metabolites are similar in human as well in pig microsomes and reconstituted systems with the isolated pig CYP1A, CYP2A19 and CYP2E1 enzyme preparations. UDP glucuronosyl transferases are the most important enzymes of the second phase of xenobiotic metabolism; experiments with two marker substrates of the form UGT1A6 have shown that these compounds are metabolized simi...
Klinicka Farmakologie a Farmacie
Objectives: Carbapenem antibiotics are used in severe nosocomial infections, so the quality of th... more Objectives: Carbapenem antibiotics are used in severe nosocomial infections, so the quality of their different preparations has been emphasized. Aim: A comparison of the quality of generic imipenem/cilastatin and meropenem produced by Fresenius Kabi with the branded preparations. Methods: 3 commercially available lots of generic preparations (Imipenem/cilastatin Kabi® and Meropenem Kabi®) were compared with one branded lot of each carbapenem (Tienam® and Meronem®). Concentrations of active agents were measured by HPLC analysis with spectrophotometric detection. Impurity or degradation product content, concentration of active agent in preparation dissolved for use, stability of dissolved preparation and plasma concentrations following intraperitoneal administration to laboratory rats were measured. Results: Concentrations of the active agents in all tested lots have not significantly varied. Impurities or degradation products were not found in any of the tested lots. All tested prepa...
Uploads
Papers by Zuzana Matušková