Alterations in the excitability of dorsal root ganglion (DRG) neurons are critical in the pathogenesis of acute and chronic pain. Neurotransmitter release from the terminals of DRG neurons is regulated by cannabinoid receptor 1 (CB1) and transient receptor potential vanilloid 1 (TRPV1), both activated by anandamide (AEA). In our experiments, the AEA precursor N-arachidonoylphosphatidylethanolamine (20:4-NAPE) was used to study the modulation of nociceptive DRG neurons excitability using K+-evoked Ca2+ transients. Intrathecal administration was used to evaluate in vivo effects. Application of 20:4-NAPE at lower concentrations (10 nM - 1 μM) decreased the excitability of DRG neurons, whereas the higher (10 μM) increased it. Both effects of 20:4-NAPE were blocked by the N-acylphosphatidylethanolamine phospholipase D (NAPE-PLD) inhibitor LEI-401. Similarly, lower concentrations of externally applied AEA (1 nM - 10 nM) inhibited DRG neurons, whereas higher concentration (100 nM) did not change it. High AEA concentration (10 μM) evoked Ca2+ transients dependent on TRPV1 activation in separate experiments. Inhibition of the CB1 receptor by PF514273 (400 nM) prevented the 20:4-NAPE- and AEA-induced inhibition, whereas TRPV1 inhibition by SB366791 (1 μM) prevented the increased DRG neuron excitability. In behavioral tests, lower 20:4-NAPE concentration caused hyposensitivity, while higher evoked mechanical allodynia. Intrathecal LEI-401 prevented both in vivo effects of 20:4-NAPE. These results highlight anti- and pro-nociceptive effects of 20:4-NAPE mediated by CB1 and TRPV1 in concentration-dependent manner. Our study underscores the complexity of endocannabinoid signaling in pain transmission modulation and highlights 20:4-NAPE as a potential therapeutic target, offering new insights for developing analgesic strategies.
- MeSH
- endokanabinoidy farmakologie metabolismus MeSH
- fosfatidylethanolaminy * farmakologie MeSH
- fosfolipasa D * metabolismus antagonisté a inhibitory MeSH
- kationtové kanály TRPV metabolismus MeSH
- krysa rodu rattus MeSH
- kyseliny arachidonové * farmakologie MeSH
- neurony * účinky léků metabolismus MeSH
- polynenasycené alkamidy farmakologie MeSH
- potkani Sprague-Dawley MeSH
- receptor kanabinoidní CB1 metabolismus MeSH
- spinální ganglia * účinky léků metabolismus cytologie MeSH
- vápník metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The thermo- and pain-sensitive Transient Receptor Potential Melastatin 3 and 8 (TRPM3 and TRPM8) ion channels are functionally associated in the lipid rafts of the plasma membrane. We have already described that cholesterol and sphingomyelin depletion, or inhibition of sphingolipid biosynthesis decreased the TRPM8 but not the TRPM3 channel opening on cultured sensory neurons. We aimed to test the effects of lipid raft disruptors on channel activation on TRPM3- and TRPM8-expressing HEK293T cells in vitro, as well as their potential analgesic actions in TRPM3 and TRPM8 channel activation involving acute pain models in mice. CHO cell viability was examined after lipid raft disruptor treatments and their effects on channel activation on channel expressing HEK293T cells by measurement of cytoplasmic Ca2+ concentration were monitored. The effects of treatments were investigated in Pregnenolone-Sulphate-CIM-0216-evoked and icilin-induced acute nocifensive pain models in mice. Cholesterol depletion decreased CHO cell viability. Sphingomyelinase and methyl-beta-cyclodextrin reduced the duration of icilin-evoked nocifensive behavior, while lipid raft disruptors did not inhibit the activity of recombinant TRPM3 and TRPM8. We conclude that depletion of sphingomyelin or cholesterol from rafts can modulate the function of native TRPM8 receptors. Furthermore, sphingolipid cleavage provided superiority over cholesterol depletion, and this method can open novel possibilities in the management of different pain conditions.
- MeSH
- analgetika farmakologie terapeutické užití MeSH
- beta-cyklodextriny * farmakologie MeSH
- bolest chemicky indukované farmakoterapie metabolismus MeSH
- CHO buňky MeSH
- cholesterol metabolismus MeSH
- Cricetulus MeSH
- HEK293 buňky MeSH
- kationtové kanály TRPM * metabolismus genetika MeSH
- lidé MeSH
- membránové mikrodomény metabolismus účinky léků MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- pregnenolon farmakologie MeSH
- pyrimidinony farmakologie MeSH
- sfingomyelinfosfodiesterasa * metabolismus farmakologie MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Rostlinné přípravky, respektive z nich vyrobené doplňky stravy, mohou vést k lékovým interakcím podobně jako léky. Výrobci takových doplňků stravy však na případné lékové interakce neupozorňují. Inhibitory fosfodiesterázy typu 5 (iPDE5) jsou široce užívané léky, jejichž společnou vlastností je odbourávání cestou CYP3A4 a CYP3A5. Inhibitory a induktory těchto isoenzymů cytochromu P-450 vyvolávají různě závažné lékové interakce a pouze k některým z nich je možné v SPC dohledat doporučený management. Cílem článku je podat základní informaci o rostlinných přípravcích, které mají potenciál ovlivnit farmakokinetické vlastnosti a v některých případech patrně i účinnost a bezpečnosti iPDE5.
Herbal products, or food supplements made from them, can lead to drug interactions as well as drugs. However, manufacturers of such food supplements do not point out possible drug interactions. Phosphodiesterase type 5 (iPDE5) inhibitors are widely used drugs, the common feature of which is degradation by the CYP3A4 and CYP3A5 pathways. Inhibitors and inducers of these cytochrome P-450 isoenzymes cause drug interactions of varying severity, and only for some of them is it possible to find a recommended management in the SPC. The aim of the article is to provide basic information about herbal products with the potential to influence pharmacokinetics and, in some cases, probably also the effectiveness and safety of iPDE5.
- Klíčová slova
- avanafil,
- MeSH
- cyklické nukleotidfosfodiesterasy, typ 5 farmakologie terapeutické užití MeSH
- erektilní dysfunkce * farmakoterapie MeSH
- inhibitory fosfodiesterasy 5 farmakologie terapeutické užití MeSH
- interakce bylin a léků * MeSH
- léčivé rostliny MeSH
- lékové interakce MeSH
- lidé MeSH
- rostlinné přípravky MeSH
- tadalafil farmakologie terapeutické užití MeSH
- vardenafil farmakologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- acyltransferasy MeSH
- arachidonát-12-lipoxygenasa genetika MeSH
- epidermis * MeSH
- fosfolipasy MeSH
- kůže MeSH
- lamelární ichtyóza * MeSH
- lipidy MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- dopisy MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
Sphingomyelinase D (SMase D), the main toxic component of Loxosceles venom, has a well-documented role on dermonecrotic lesion triggered by envenomation with these species; however, the intracellular mechanisms involved in this event are still poorly known. Through differential transcriptomics of human keratinocytes treated with L. laeta or L. intermedia SMases D, we identified 323 DEGs, common to both treatments, as well as upregulation of molecules involved in the IL-1 and ErbB signaling. Since these pathways are related to inflammation and wound healing, respectively, we investigated the relative expression of some molecules related to these pathways by RT-qPCR and observed different expression profiles over time. Although, after 24 h of treatment, both SMases D induced similar modulation of these pathways in keratinocytes, L. intermedia SMase D induced earlier modulation compared to L. laeta SMase D treatment. Positive expression correlations of the molecules involved in the IL-1 signaling were also observed after SMases D treatment, confirming their inflammatory action. In addition, we detected higher relative expression of the inhibitor of the ErbB signaling pathway, ERRFI1, and positive correlations between this molecule and pro-inflammatory mediators after SMases D treatment. Thus, herein, we describe the cell pathways related to the exacerbation of inflammation and to the failure of the wound healing, highlighting the contribution of the IL-1 signaling pathway and the ERRFI1 for the development of cutaneous loxoscelism.
- MeSH
- erbB receptory metabolismus MeSH
- fosfodiesterasy toxicita MeSH
- interleukin-1 metabolismus MeSH
- kousnutí pavoukem patologie MeSH
- lidé MeSH
- pavoučí jedy * toxicita MeSH
- pavouci chemie metabolismus MeSH
- sfingomyelinfosfodiesterasa * metabolismus MeSH
- signální transdukce MeSH
- zánět MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Neutral sphingomyelinase 2 (nSMase2) has gained increasing attention as a therapeutic target to regulate ceramide production in various disease conditions. Phenyl (R)-(1-(3-(3,4-dimethoxyphenyl)-2,6-dimethylimidazo[1,2-b]pyridazin-8-yl)-pyrrolidin-3-yl)carbamate (PDDC) is a submicromolar nSMase2 inhibitor and has been widely used to study the pharmacological effects of nSMase2 inhibition. Through screening of compounds containing a bicyclic 5-6 fused ring, larotrectinib containing a pyrazolo[1,5-a]pyrimidine ring was identified as a low micromolar inhibitor of nSMase2. This prompted us to investigate the pyrazolo[1,5-a]pyrimidin-3-amine ring as a novel scaffold to replace the imidazo[1,2-b]pyridazine-8-amine ring of PDDC. A series of molecules containing a pyrazolo[1,5-a]pyrimidin-3-amine ring were synthesized and tested for their ability to inhibit human nSMase2. Several compounds exhibited nSMase2 inhibitory potency superior to that of PDDC. Among these, N,N-dimethyl-5-morpholinopyrazolo[1,5-a]pyrimidin-3-amine (11j) was found to be metabolically stable in liver microsomes and orally available with a favorable brain-to-plasma ratio, demonstrating the potential of pyrazolo[1,5-a]pyrimidine ring as an effective scaffold for nSMase2 inhibition.
- MeSH
- aminy * MeSH
- ceramidy MeSH
- lidé MeSH
- pyrimidiny farmakologie MeSH
- sfingomyelinfosfodiesterasa * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Rostlinné přípravky, respektive z nich vyrobené doplňky stravy, mohou vést k lékovým interakcím podobně jako léky. Výrobci takových doplňků stravy však na případné lékové interakce neupozorňují. Inhibitory fosfodiesterázy typu 5 (iPDE5) jsou široce užívané léky, jejichž společnou vlastností je odbourávání cestou CYP3A4 a CYP3A5. Inhibitory a induktory těchto isoenzymů cytochromu P-450 vyvolávají různě závažné lékové interakce a pouze k některým z nich je možné v SPC dohledat doporučený management. Cílem článku je podat základní informaci o rostlinných přípravcích, které mají potenciál ovlivnit farmakokinetické vlastnosti a v některých případech patrně i účinnost a bezpečnosti iPDE5.
Herbal products, or food supplements made from them, can lead to drug interactions as well as drugs. However, manufacturers of such food supplements do not point out possible drug interactions. Phosphodiesterase type 5 (iPDE5) inhibitors are widely used drugs, the common feature of which is degradation by the CYP3A4 and CYP3A5 pathways. Inhibitors and inducers of these cytochrome P-450 isoenzymes cause drug interactions of varying severity, and only for some of them is it possible to find a recommended management in the SPC. The aim of the article is to provide basic information about herbal products with the potential to influence pharmacokinetics and, in some cases, probably also the effectiveness and safety of iPDE5.
- Klíčová slova
- avanafil,
- MeSH
- cyklické nukleotidfosfodiesterasy, typ 5 farmakologie terapeutické užití MeSH
- erektilní dysfunkce * farmakoterapie MeSH
- inhibitory fosfodiesterasy 5 farmakologie terapeutické užití MeSH
- interakce bylin a léků * MeSH
- léčivé rostliny MeSH
- lékové interakce MeSH
- lidé MeSH
- rostlinné přípravky MeSH
- tadalafil farmakologie terapeutické užití MeSH
- vardenafil farmakologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- přehledy MeSH
BACKGROUND: DNA-protein cross-links (DPCs) are one of the most deleterious DNA lesions, originating from various sources, including enzymatic activity. For instance, topoisomerases, which play a fundamental role in DNA metabolic processes such as replication and transcription, can be trapped and remain covalently bound to DNA in the presence of poisons or nearby DNA damage. Given the complexity of individual DPCs, numerous repair pathways have been described. The protein tyrosyl-DNA phosphodiesterase 1 (Tdp1) has been demonstrated to be responsible for removing topoisomerase 1 (Top1). Nevertheless, studies in budding yeast have indicated that alternative pathways involving Mus81, a structure-specific DNA endonuclease, could also remove Top1 and other DPCs. RESULTS: This study shows that MUS81 can efficiently cleave various DNA substrates modified by fluorescein, streptavidin or proteolytically processed topoisomerase. Furthermore, the inability of MUS81 to cleave substrates bearing native TOP1 suggests that TOP1 must be either dislodged or partially degraded prior to MUS81 cleavage. We demonstrated that MUS81 could cleave a model DPC in nuclear extracts and that depletion of TDP1 in MUS81-KO cells induces sensitivity to the TOP1 poison camptothecin (CPT) and affects cell proliferation. This sensitivity is only partially suppressed by TOP1 depletion, indicating that other DPCs might require the MUS81 activity for cell proliferation. CONCLUSIONS: Our data indicate that MUS81 and TDP1 play independent roles in the repair of CPT-induced lesions, thus representing new therapeutic targets for cancer cell sensitisation in combination with TOP1 inhibitors.
- MeSH
- DNA vazebné proteiny * genetika metabolismus MeSH
- DNA-topoisomerasy I genetika metabolismus MeSH
- endonukleasy * genetika metabolismus MeSH
- fosfodiesterasy * genetika metabolismus MeSH
- oprava DNA MeSH
- poškození DNA MeSH
- Saccharomyces cerevisiae - proteiny * genetika metabolismus MeSH
- Saccharomyces cerevisiae MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Autotaxin, also known as ecto-nucleotide pyrophosphatase/phosphodiesterase family member 2, is a secreted glycoprotein that plays multiple roles in human physiology and cancer pathology. This protein, by converting lysophosphatidylcholine into lysophosphatidic acid, initiates a complex signalling cascade with significant biological implications. The article outlines the autotaxin gene and protein structure, expression regulation and physiological functions, but focuses mainly on the role of autotaxin in cancer development and progression. Autotaxin and lysophosphatidic acid signalling influence several aspects of cancer, including cell proliferation, migration, metastasis, therapy resistance, and interactions with the immune system. The potential of autotaxin as a diagnostic biomarker and promising drug target is also examined.
- MeSH
- fosfodiesterasy * genetika chemie metabolismus MeSH
- lidé MeSH
- lysofosfolipidy metabolismus MeSH
- nádory * MeSH
- signální transdukce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Niemann‐Pickova choroba typu B (NPB) představuje vzácnou dědičnou metabolickou chorobu, která má velmi široké spektrum příznaků s různým věkem nástupu a tíží. Přesná diagnostika je založena na enzymologickém a současně genetickém vyšetření. V současné době je již dostupná specifická léčba pomocí enzymové substituční terapie olipudázou alfa. V tomto článku prezentujeme dospělého pacienta s náhodným nálezem splenomegalie, u kterého byla následně potvrzena diagnóza NPB.
Niemann-Pick disease type B (NPB) is a rare inherited metabolic disease that has a very wide spectrum of symptoms with different ages of onset and severity. Accurate diagnosis is based on an enzymological and genetic examination. Currently, a specific treatment using enzyme replacement therapy with olipudase alfa is already available. In this article, we present an adult patient with a random finding of splenomegaly, in whom the diagnosis of NPB was subsequently confirmed.
