Albendazole (ABZ) is an anthelmintic frequently used to treat haemonchosis, a common parasitosis of ruminants caused by the gastrointestinal nematode Haemonchus contortus. This parasite is able to protect itself against ABZ via the formation of inactive ABZ-glycosides. The present study was designed to deepen the knowledge about the role of UDP-glycosyltransferases (UGTs) in ABZ glycosylation in H. contortus. The induction effect of phenobarbital, a classical inducer of UGTs, as well as ABZ and ABZ-sulphoxide (ABZSO, the main active metabolite of ABZ) on UGTs expression and UGT activity toward ABZ was studied ex vivo in isolated adult nematodes. The effect of three potential UGT inhibitors (5-nitrouracil, 4,6-dihydroxy-5-nitropyrimidine and sulfinpyrazone) on ABZ glycosylation was tested. Pre-incubation of nematodes with ABZ and ABZSO led to increased expression of several UGTs as well as ABZ-glycosides formation in subsequent treatment. Phenobarbital also induced UGTs expression, but did not affect ABZ biotransformation. In the nematode's subcellular fraction, sulfinpyrazone inhibited UGT activity toward ABZ, although no effect of other inhibitors was observed. The inhibitory potential of sulfinpyrazone on the formation of ABZ-glycosides was also proved ex vivo in living nematodes. The obtained results confirmed the role of UGTs in ABZ biotransformation in H. contortus adults and revealed sulfinpyrazone as a potent inhibitor of ABZ glycosylation in this parasite. The possible use of sulfinpyrazone with ABZ in combination therapy merits further research.
- Klíčová slova
- Anthelmintic resistance, Anthelmintics biotransformation, Benzimidazoles, Detoxification, Gene expression, Glycosylated metabolites, Glycosylation, Nematodes, UGT inhibitors, UHPLC-MS/MS,
- MeSH
- albendazol MeSH
- anthelmintika * terapeutické užití MeSH
- fenobarbital metabolismus farmakologie terapeutické užití MeSH
- glykosidy metabolismus farmakologie terapeutické užití MeSH
- glykosyltransferasy MeSH
- Haemonchus * MeSH
- hlístice * MeSH
- nemoci ovcí * farmakoterapie MeSH
- ovce MeSH
- sulfinpyrazon metabolismus farmakologie terapeutické užití MeSH
- uridindifosfát MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- albendazol MeSH
- anthelmintika * MeSH
- fenobarbital MeSH
- glykosidy MeSH
- glykosyltransferasy MeSH
- sulfinpyrazon MeSH
- uridindifosfát MeSH
Veterinary anthelmintics excreted from treated animals pass to soil, subsequently to plants and then to their consumers. This circulation might have various consequences, including drug-resistance promotion in helminths. The present study was designed to follow the effect of the environmental circulation of the common anthelmintic drug albendazole (ABZ) in real farm conditions on the parasitic nematode Haemonchus contortus in vivo. Two fields with fodder plants (clover and alfalfa) were fertilized, the first with dung from ABZ-treated sheep (at the recommended dosage), the second with dung from non-treated sheep (controls). After a 10-week growth period, the fresh fodder from both fields was used to feed two groups of sheep, which were infected with H. contortus. Eggs and adult nematodes from the animals of both groups were isolated, and various parameters were compared. No significant changes in the eggs' sensitivity to ABZ and thiabendazole were observed. However, significantly increased expression of several cytochromes P450 and UDP-glycosyl transferases as well as increased oxidation and glycosylation of ABZ and ABZ-sulfoxide (ABZ-SO) was found in the exposed nematodes. These results show that ABZ environmental circulation improves the ability of the helminths to deactivate ABZ.
- Klíčová slova
- Cytochromes P450, Drug resistance, Efflux transporters, Micropollutants, UDP-glycosyl transferases, Veterinary drugs,
- MeSH
- albendazol metabolismus farmakologie terapeutické užití MeSH
- anthelmintika * metabolismus farmakologie terapeutické užití MeSH
- Haemonchus * metabolismus MeSH
- hlístice * MeSH
- léková rezistence MeSH
- ovce MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- albendazol MeSH
- anthelmintika * MeSH
BACKGROUND: MAPK/ERK signaling is a well-known mediator of extracellular stimuli controlling intracellular responses to growth factors and mechanical cues. The critical requirement of MAPK/ERK signaling for embryonic stem cell maintenance is demonstrated, but specific functions in progenitor regulation during embryonic development, and in particular kidney development remain largely unexplored. We previously demonstrated MAPK/ERK signaling as a key regulator of kidney growth through branching morphogenesis and normal nephrogenesis where it also regulates progenitor expansion. Here, we performed RNA sequencing-based whole-genome expression analysis to identify transcriptional MAPK/ERK targets in two distinct renal populations: the ureteric bud epithelium and the nephron progenitors. RESULTS: Our analysis revealed a large number (5053) of differentially expressed genes (DEGs) in nephron progenitors and significantly less (1004) in ureteric bud epithelium, reflecting likely heterogenicity of cell types. The data analysis identified high tissue-specificity, as only a fraction (362) of MAPK/ERK targets are shared between the two tissues. Tissue-specific MAPK/ERK targets participate in the regulation of mitochondrial energy metabolism in nephron progenitors, which fail to maintain normal mitochondria numbers in the MAPK/ERK-deficient tissue. In the ureteric bud epithelium, a dramatic decline in progenitor-specific gene expression was detected with a simultaneous increase in differentiation-associated genes, which was not observed in nephron progenitors. Our experiments in the genetic model of MAPK/ERK deficiency provide evidence that MAPK/ERK signaling in the ureteric bud maintains epithelial cells in an undifferentiated state. Interestingly, the transcriptional targets shared between the two tissues studied are over-represented by histone genes, suggesting that MAPK/ERK signaling regulates cell cycle progression and stem cell maintenance through chromosome condensation and nucleosome assembly. CONCLUSIONS: Using tissue-specific MAPK/ERK inactivation and RNA sequencing in combination with experimentation in embryonic kidneys, we demonstrate here that MAPK/ERK signaling maintains ureteric bud tip cells, suggesting a regulatory role in collecting duct progenitors. We additionally deliver new mechanistic information on how MAPK/ERK signaling regulates progenitor maintenance through its effects on chromatin accessibility and energy metabolism.
A combination of several pharmacophores in one molecule has been successfully used for multi-target-directed ligands (MTDL) design. New propargylamine substituted derivatives combined with salicylic and cinnamic scaffolds were designed and synthesized as potential cholinesterases and monoamine oxidases (MAOs) inhibitors. They were evaluated invitro for inhibition of acetyl- (AChE) and butyrylcholinesterase (BuChE) using Ellman's method. All the compounds act as dual inhibitors. Most of the derivatives are stronger inhibitors of AChE, the best activity showed 5-bromo-N-(prop-2-yn-1-yl)salicylamide 1e (IC50 = 8.05 µM). Carbamates (4-bromo-2-[(prop-2-yn-1-yl)carbamoyl]phenyl ethyl(methyl)carbamate 2d and 2,4-dibromo-6-[(prop-2-yn-1-yl)carbamoyl]phenyl ethyl(methyl)carbamate 2e were selective and the most active for BuChE (25.10 and 26.09 µM). 4-Bromo-2-[(prop-2-yn-1-ylimino)methyl]phenol 4a was the most potent inhibitor of MAOs (IC50 of 3.95 and ≈10 µM for MAO-B and MAO-A, respectively) along with a balanced inhibition of both cholinesterases being a real MTDL. The mechanism of action was proposed, and binding modes of the hits were studied by molecular docking on human enzymes. Some of the derivatives also exhibited antioxidant properties. Insilico prediction of physicochemical parameters affirm that the molecules would be active after oral administration and able to reach brain tissue.
- Klíčová slova
- Acetylcholinesterase, Butyrylcholinesterase, Enzyme inhibition, Molecular docking, Monoamine oxidases, Multitargeting ligands, Propargylamine, Salicylic scaffold,
- MeSH
- antioxidancia chemická syntéza chemie farmakologie MeSH
- butyrylcholinesterasa metabolismus MeSH
- cholinesterasové inhibitory chemická syntéza chemie farmakologie MeSH
- cholinesterasy metabolismus MeSH
- Electrophorus MeSH
- hepatocyty účinky léků metabolismus MeSH
- inhibitory MAO chemická syntéza chemie farmakologie MeSH
- koně MeSH
- krysa rodu Rattus MeSH
- kultivované buňky MeSH
- lidé MeSH
- molekulární struktura MeSH
- monoaminoxidasa metabolismus MeSH
- pargylin analogy a deriváty chemická syntéza chemie farmakologie MeSH
- potkani Wistar MeSH
- propylaminy chemická syntéza chemie farmakologie MeSH
- reaktivní formy kyslíku metabolismus MeSH
- simulace molekulového dockingu * MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antioxidancia MeSH
- butyrylcholinesterasa MeSH
- cholinesterasové inhibitory MeSH
- cholinesterasy MeSH
- inhibitory MAO MeSH
- monoaminoxidasa MeSH
- pargylin MeSH
- propargylamine MeSH Prohlížeč
- propylaminy MeSH
- reaktivní formy kyslíku MeSH
Anthelmintics, drugs against parasitic worms, are frequently used in livestock and might act as danger environmental microcontaminants. The present study was designed to monitor the possible circulation of common anthelmintic drug albendazole (ABZ) and its metabolites in the real agriculture conditions. The sheep were treated with the recommended dose of ABZ. Collected faeces were used for the fertilization of a field with fodder plants (alfalfa and clover) which served as feed for sheep from a different farm. The selective ultrasensitive mass spectrometry revealed surprisingly high concentrations of active ABZ metabolite (ABZ-sulphoxide) in all samples (dung, plants, ovine plasma, rumen content and faeces). Our results prove for the first time an undesirable permeation of ABZ metabolites from sheep excrement into plants (used as fodder) and subsequently to other sheep in real agricultural conditions. This circulation causes the permanent exposition of the ecosystems and food-chain to the drug and can promote the development of drug resistance in helminths.
- Klíčová slova
- Anthelmintics, Benzimidazoles, Drug resistance, Fodder plant, Micropollutants,
- MeSH
- albendazol MeSH
- anthelmintika * MeSH
- ekosystém MeSH
- farmy MeSH
- ovce MeSH
- veterinární léky * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- albendazol MeSH
- anthelmintika * MeSH
- veterinární léky * MeSH
Glutathione peroxidase 7 (GPx7) acts as an intracellular stress sensor/transmitter and plays an important role in adipocyte differentiation and the prevention of obesity related pathologies. For this reason, finding the regulatory mechanisms that control GPx7 expression is of great importance. As microRNAs (miRNAs) could participate in the regulation of GPx7 expression, we studied the inhibition of GPx7 expression by four selected miRNAs with relation to obesity and adipogenesis. The effect of the transfection of selected miRNAs mimics on GPx7 expression was tested in three cell models (HEK293, SW480, AT-MSC). The interaction of selected miRNAs with the 3'UTR of GPx7 was followed up on using a luciferase gene reporter assay. In addition, the levels of GPx7 and selected miRNAs in adipose tissue mesenchymal stem cells (AT-MSC) and mature adipocytes from four human donors were compared, with the changes in these levels during adipogenesis analyzed. Our results show for the first time that miR-137 and miR-29b bind to the 3'UTR region of GPx7 and inhibit the expression of this enzyme at the mRNA and protein level in all the human cells tested. However, no negative correlation between miR-137 nor miR-29b level and GPx7 was observed during adipogenesis. Despite the confirmed inhibition of GPx7 expression by miR-137 and miR-29b, the action of these two molecules in adipogenesis and mature adipocytes must be accompanied by other regulators.
- Klíčová slova
- Gene regulation, Glutathione peroxidase, miRNA,
- MeSH
- 3' nepřekládaná oblast MeSH
- adipogeneze genetika MeSH
- glutathionperoxidasa MeSH
- kmenové buňky metabolismus MeSH
- kultivované buňky MeSH
- lidé středního věku MeSH
- lidé MeSH
- messenger RNA metabolismus MeSH
- mikro RNA metabolismus MeSH
- nádorové buněčné linie MeSH
- peroxidasy genetika MeSH
- regulace genové exprese enzymů * MeSH
- tukové buňky metabolismus MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 3' nepřekládaná oblast MeSH
- glutathionperoxidasa MeSH
- GPX7 protein, human MeSH Prohlížeč
- messenger RNA MeSH
- mikro RNA MeSH
- peroxidasy MeSH
Parasitic nematodes transition between dramatically different free-living and parasitic stages, with correctly timed development and migration crucial to successful completion of their lifecycle. However little is known of the mechanisms controlling these transitions. microRNAs (miRNAs) negatively regulate gene expression post-transcriptionally and regulate development of diverse organisms. Here we used microarrays to determine the expression profile of miRNAs through development and in gut tissue of the pathogenic nematode Haemonchus contortus. Two miRNAs, mir-228 and mir-235, were enriched in infective L3 larvae, an arrested stage analogous to Caenorhabditis elegans dauer larvae. We hypothesized that these miRNAs may suppress development and maintain arrest. Consistent with this, inhibitors of these miRNAs promoted H. contortus development from L3 to L4 stage, while genetic deletion of C. elegans homologous miRNAs reduced dauer arrest. Epistasis studies with C. elegans daf-2 mutants showed that mir-228 and mir-235 synergise with FOXO transcription factor DAF-16 in the insulin signaling pathway. Target prediction suggests that these miRNAs suppress metabolic and transcription factor activity required for development. Our results provide novel insight into the expression and functions of specific miRNAs in regulating nematode development and identify miRNAs and their target genes as potential therapeutic targets to limit parasite survival within the host.
- MeSH
- Caenorhabditis elegans genetika MeSH
- cholesteny farmakologie MeSH
- delece genu MeSH
- druhová specificita MeSH
- genová ontologie MeSH
- Haemonchus účinky léků genetika růst a vývoj MeSH
- larva MeSH
- messenger RNA genetika metabolismus MeSH
- mikro RNA biosyntéza genetika MeSH
- proteiny Caenorhabditis elegans genetika MeSH
- receptor inzulinu genetika MeSH
- RNA helmintů biosyntéza genetika MeSH
- vývojová regulace genové exprese účinky léků MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cholesteny MeSH
- DAF-2 protein, C elegans MeSH Prohlížeč
- dafachronic acid MeSH Prohlížeč
- messenger RNA MeSH
- mikro RNA MeSH
- proteiny Caenorhabditis elegans MeSH
- receptor inzulinu MeSH
- RNA helmintů MeSH
The selection of a suitable combination of reference genes (RGs) for data normalization is a crucial step for obtaining reliable and reproducible results from transcriptional response analysis using a reverse transcription-quantitative polymerase chain reaction. This is especially so if a three-dimensional multicellular model prepared from liver tissues originating from biologically diverse human individuals is used. The mRNA and miRNA RGs stability were studied in thirty-five human liver tissue samples and twelve precision-cut human liver slices (PCLS) treated for 24 h with dimethyl sulfoxide (controls) and PCLS treated with β-naphthoflavone (10 µM) or rifampicin (10 µM) as cytochrome P450 (CYP) inducers. Validation of RGs was performed by an expression analysis of CYP3A4 and CYP1A2 on rifampicin and β-naphthoflavone induction, respectively. Regarding mRNA, the best combination of RGs for the controls was YWHAZ and B2M, while YWHAZ and ACTB were selected for the liver samples and treated PCLS. Stability of all candidate miRNA RGs was comparable or better than that of generally used short non-coding RNA U6. The best combination for the control PCLS was miR-16-5p and miR-152-3p, in contrast to the miR-16-5b and miR-23b-3p selected for the treated PCLS. Our results showed that the candidate RGs were rather stable, especially for miRNA in human PCLS.
- Klíčová slova
- RT-qPCR, human liver, mRNA, miRNA, precision-cut liver slices, reference gene,
- MeSH
- beta-2-mikroglobulin genetika metabolismus MeSH
- beta-naftoflavon farmakologie MeSH
- cytochrom P-450 CYP1A2 genetika metabolismus MeSH
- cytochrom P-450 CYP3A genetika metabolismus MeSH
- dimethylsulfoxid farmakologie MeSH
- dospělí MeSH
- játra účinky léků metabolismus MeSH
- kvantitativní polymerázová řetězová reakce normy MeSH
- lidé středního věku MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- mikro RNA genetika metabolismus MeSH
- proteiny 14-3-3 genetika metabolismus MeSH
- referenční standardy MeSH
- rifampin farmakologie MeSH
- senioři MeSH
- stanovení celkové genové exprese normy MeSH
- systém (enzymů) cytochromů P-450 farmakologie MeSH
- transkriptom MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- B2M protein, human MeSH Prohlížeč
- beta-2-mikroglobulin MeSH
- beta-naftoflavon MeSH
- CYP1A2 protein, human MeSH Prohlížeč
- CYP3A4 protein, human MeSH Prohlížeč
- cytochrom P-450 CYP1A2 MeSH
- cytochrom P-450 CYP3A MeSH
- dimethylsulfoxid MeSH
- messenger RNA MeSH
- mikro RNA MeSH
- proteiny 14-3-3 MeSH
- rifampin MeSH
- systém (enzymů) cytochromů P-450 MeSH
- YWHAZ protein, human MeSH Prohlížeč
High fructose intake from soft drinks and sweets is assumed to have a negative impact on human health. Yet in spite of intensive research, the molecular mechanisms of these effects have not been fully elucidated yet, for example, the effect of high fructose intake could be different in normal and obese individuals. Four groups of mice were used in this study: control groups of lean mice and mice with obesity induced by a high-fat diet, then both of these groups with or without fructose administration in drinks. In plasma of each group, triacylglycerol, cholesterol, free fatty acids, alanine aminotransferase, insulin and adiponectin were measured. The expression levels of selected microRNAs (miRNAs) in plasma, the liver, white adipose tissue, brown adipose tissue and subcutaneous adipose tissue were quantified. In both lean and obese mice, high fructose intake increased cholesterol amount in the liver, up-regulated hepatic miR-27a, down-regulated miR-33a in white adipose tissue and increased plasmatic level of miR-21. The effect of high fructose intake on other miRNAs in the liver, plasma and adipose tissues differed in normal and obese mice. Fructose intake led to hepatic hypercholesterolemia and aberrant expression of several miRNAs participating in lipid metabolism, adipocytes differentiation and nonalcoholic fatty liver disease promotion. The effect of fructose on miRNAs expression differed in normal and obese mice. Nevertheless, plasmatic miR-21, which was induced by fructose in both lean and obese mice, may be considered as a potential biomarker of excessive fructose intake.
- Klíčová slova
- Fructose, MicroRNA, Obesity, adipose tissue, lipid metabolism, miR-21,
- MeSH
- bílá tuková tkáň účinky léků fyziologie MeSH
- dieta s vysokým obsahem tuků škodlivé účinky MeSH
- fruktosa škodlivé účinky MeSH
- hnědá tuková tkáň účinky léků fyziologie MeSH
- játra účinky léků fyziologie MeSH
- mikro RNA krev účinky léků MeSH
- myši inbrední C57BL MeSH
- obezita genetika metabolismus MeSH
- regulace genové exprese účinky léků MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fruktosa MeSH
- mikro RNA MeSH
Beer, the most popular beverage containing hops, is also frequently consumed by cancer patients. Moreover, non-alcoholic beer, owing to its nutritional value and high content of biological active compounds, is sometimes recommended to patients by oncologists. However, the potential benefits and negatives have to date not been sufficiently evaluated. The present study was designed to examine the effects of four main hop-derived prenylflavonoids on the viability, reactive oxygen species (ROS) formation, activity of caspases, and efficiency of the chemotherapeutics 5-fluorouracil (5-FU), oxaliplatin (OxPt) and irinotecan (IRI) in colorectal cancer cell lines SW480, SW620 and CaCo-2. All the prenylflavonoids exerted substantial antiproliferative effects in all cell lines, with xanthohumol being the most effective (IC50 ranging from 3.6 to 7.3 µM). Isoxanthohumol increased ROS formation and the activity of caspases-3/7, but 6-prenylnaringenin and 8-prenylnaringenin exerted antioxidant properties. As 6-prenylnaringenin acted synergistically with IRI, its potential in combination therapy deserves further study. However, other prenylflavonoids acted antagonistically with all chemotherapeutics at least in one cell line. Therefore, consumption of beer during chemotherapy with 5-FU, OxPt and IRI should be avoided, as the prenylflavonoids in beer could decrease the efficacy of the treatment.
- Klíčová slova
- 5-fluorouracil, caspase activity, colorectal carcinoma cells, irinotecan, isoxanthohumol, naringenin, oxaliplatin, prenylflavonoids,
- MeSH
- antioxidancia MeSH
- Caco-2 buňky MeSH
- fixní kombinace léků MeSH
- flavanony farmakologie terapeutické užití MeSH
- flavonoidy farmakologie terapeutické užití MeSH
- fluoruracil terapeutické užití MeSH
- fytogenní protinádorové látky farmakologie terapeutické užití MeSH
- Humulus chemie MeSH
- irinotekan terapeutické užití MeSH
- kaspasy metabolismus MeSH
- kolorektální nádory farmakoterapie metabolismus MeSH
- lékové interakce * MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- oxaliplatin terapeutické užití MeSH
- pivo * škodlivé účinky MeSH
- propiofenony farmakologie terapeutické užití MeSH
- protinádorové látky farmakologie terapeutické užití MeSH
- reaktivní formy kyslíku metabolismus MeSH
- rostlinné extrakty farmakologie terapeutické užití MeSH
- stravovací zvyklosti MeSH
- výsledek terapie MeSH
- xantony farmakologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 6-prenylnaringenin MeSH Prohlížeč
- 8-prenylnaringenin MeSH Prohlížeč
- antioxidancia MeSH
- fixní kombinace léků MeSH
- flavanony MeSH
- flavonoidy MeSH
- fluoruracil MeSH
- fytogenní protinádorové látky MeSH
- irinotekan MeSH
- isoxanthohumol MeSH Prohlížeč
- kaspasy MeSH
- oxaliplatin MeSH
- propiofenony MeSH
- protinádorové látky MeSH
- reaktivní formy kyslíku MeSH
- rostlinné extrakty MeSH
- xanthohumol MeSH Prohlížeč
- xantony MeSH
