P2Y12 receptors on the platelet plasma membrane are targeted by several antiplatelets drugs. Although oligomerization and functioning of P2Y12 receptors depend on the membrane environment, little is known about their preferred membrane localization and the role of surrounding lipid composition, especially the arachidonic acids (ARA), which are abundant in platelets. Coarse-grained molecular dynamics simulations of platelet plasma membrane based on the lipidomics data were used to investigate the P2Y12 lipid environment and the involvement of ARA in its oligomerization in platelet plasma membranes. The platelet plasma membrane contains two types of lipids nanodomains: ordered, enriched in SM and cholesterol, and disordered, enriched in ARA-containing lipids. P2Y12 receptors prefer to localize in these ARA-rich domains and induce the sorting of the ARA-containing lipids in their vicinity. This ARA-rich environment promotes the oligomerization of P2Y12 receptors and stabilizes the protein-protein interfaces of oligomers. As summary, oligomerization of P2Y12 receptors is promoted in ARA-rich nano-domains of the platelet plasma membrane.

• Klíčová slova
• Arachidonic acid, Blood platelets, Molecular dynamics simulation, Protein Multimerization, Protein–lipid interaction, Purinergic receptor P2Y12,
• MeSH
• buněčná membrána * metabolismus   chemie   MeSH
• cholesterol metabolismus   chemie   MeSH
• kyselina arachidonová * metabolismus   chemie   MeSH
• lidé MeSH
• multimerizace proteinu MeSH
• purinergní receptory P2Y12 * metabolismus   chemie   MeSH
• simulace molekulární dynamiky * MeSH
• trombocyty * metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cholesterol MeSH
• kyselina arachidonová * MeSH
• P2RY12 protein, human MeSH Prohlížeč
• purinergní receptory P2Y12 * MeSH

Metformin can reduce cardiovascular risk independent of glycemic control. The mechanisms behind its non-glycemic benefits, which include decreased energy intake, lower blood pressure and improved lipid and fatty acid metabolism, are not fully understood. In our study, metformin treatment reduced myocardial accumulation of neutral lipids-triglycerides, cholesteryl esters and the lipotoxic intermediates-diacylglycerols and lysophosphatidylcholines in a prediabetic rat model (p < 0.001). We observed an association between decreased gene expression and SCD-1 activity (p < 0.05). In addition, metformin markedly improved phospholipid fatty acid composition in the myocardium, represented by decreased SFA profiles and increased n3-PUFA profiles. Known for its cardioprotective and anti-inflammatory properties, metformin also had positive effects on arachidonic acid metabolism and CYP-derived arachidonic acid metabolites. We also found an association between increased gene expression of the cardiac isoform CYP2c with increased 14,15-EET (p < 0.05) and markedly reduced 20-HETE (p < 0.001) in the myocardium. Based on these results, we conclude that metformin treatment reduces the lipogenic enzyme SCD-1 and the accumulation of the lipotoxic intermediates diacylglycerols and lysophosphatidylcholine. Increased CYP2c gene expression and beneficial effects on CYP-derived arachidonic acid metabolites in the myocardium can also be involved in cardioprotective effect of metformin.

• Klíčová slova
• arachidonic acid, cytochrome P450, fatty acid profile, lipotoxic intermediates, metformin, myocardial function, myocardial phospholipids, stearoyl-CoA desaturase,
• MeSH
• bazální metabolismus účinky léků   MeSH
• biologické markery krev   MeSH
• desaturasy mastných kyselin metabolismus   MeSH
• hyperlipoproteinemie typ IV farmakoterapie   metabolismus   MeSH
• hypoglykemika farmakologie   MeSH
• kardiotonika farmakologie   MeSH
• krysa rodu Rattus MeSH
• kyselina arachidonová metabolismus   MeSH
• mediátory zánětu krev   MeSH
• metabolismus lipidů účinky léků   MeSH
• metformin farmakologie   MeSH
• modely nemocí na zvířatech MeSH
• myokard metabolismus   MeSH
• potkani Wistar MeSH
• prediabetes farmakoterapie   metabolismus   MeSH
• rizikové faktory MeSH
• srdce účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• desaturasy mastných kyselin MeSH
• hypoglykemika MeSH
• kardiotonika MeSH
• kyselina arachidonová MeSH
• mediátory zánětu MeSH
• metformin MeSH

Flavonoids, important components of human diet, have been claimed to possess a significant antiplatelet potential, in particular due to their effects on the arachidonic acid cascade. Due to variable and incomplete results, this study was aimed at delivering a detailed analysis of the effects of 29 structurally relevant, mainly natural flavonoids on three consecutive steps of the arachidonic acid cascade.Only the isoflavonoids genistein and daidzein were shown to possess a marked cyclooxygenase-1 inhibitory activity, which was higher than that of acetylsalicylic acid using the isolated ovine enzyme, and physiologically relevant, although lower than acetylsalicylic acid in human platelets. None of the tested flavonoids possesses an effect on thromboxane synthase in a clinically achievable concentration. Contrarily, many flavonoids, particularly those possessing an isolated 7-hydroxyl group and/or a 4'-hydroxyl group, acted as antagonists on thromboxane receptors. Interestingly, the substitution of the free 7-hydroxyl group by glucose might not abolish the activity.In conclusion, the consumption of few flavonoids in a diet, particularly of the isoflavonoids genistein and daidzein, may positively influence platelet aggregation.

• MeSH
• cyklooxygenasa 1 metabolismus   MeSH
• flavonoidy chemie   farmakologie   MeSH
• inhibitory agregace trombocytů chemie   farmakologie   MeSH
• inhibitory cyklooxygenasy farmakologie   MeSH
• kyselina arachidonová antagonisté a inhibitory   metabolismus   MeSH
• lidé MeSH
• receptory thromboxanů antagonisté a inhibitory   MeSH
• thromboxan-A-synthasa antagonisté a inhibitory   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cyklooxygenasa 1 MeSH
• flavonoidy MeSH
• inhibitory agregace trombocytů MeSH
• inhibitory cyklooxygenasy MeSH
• kyselina arachidonová MeSH
• receptory thromboxanů MeSH
• thromboxan-A-synthasa MeSH

BACKGROUND: Various compounds that inhibit processing of arachidonic acid (AA) are being intensively tested for their antitumour activity. However, the mechanisms responsible for such activity remain rather elusive. To approach this issue, we examined the effects of several structurally different inhibitors of AA metabolism in the human keratinocyte HaCaT cell line. METHODS: Several parameters were determined in HaCaT cells exposed to increasing concentrations of the inhibitors for 24 and/or 48 h. These included (1) oxidoreductase activity, total protein mass and cell cycle distribution to assess cell proliferation, (2) degradation of PARP protein to assess apoptosis, and (3) cell morphology, distribution of F-actin and expression of cytokeratins and E-cadherin to evaluate changes in differentiation status. RESULTS: While eicosatetraynoic acid (ETYA), nordihydroguaiaretic acid (NDGA), esculetin and MK-886 reduced proliferation of HaCaT cells, the cyclooxygenase inhibitors indomethacin and piroxicam had no such effects. Esculetin and NDGA arrested cells in S phase, and ETYA and MK-886 delayed cell progression through G(1) phase. Higher concentrations of NDGA, MK886 and/or ETYA caused cleavage of PARP. No changes in the expression of cytokeratins and E-cadherin were observed upon treatment with any of the inhibitors. However, esculetin induced redistribution of F-actin accompanied by increased cell adhesion and size. CONCLUSION: Our findings indicate that, in addition to their ability to inhibit cell proliferation and to induce apoptosis, lipoxygenase inhibitors and/or ETYA may also elicit other important physiological responses in HaCaT keratinocytes.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčné dělení účinky léků   MeSH
• indoly farmakologie   MeSH
• inhibitory cyklooxygenasy farmakologie   MeSH
• inhibitory lipoxygenas farmakologie   MeSH
• keratinocyty cytologie   účinky léků   fyziologie   MeSH
• kyselina 5,8,11,14-eikosatetraenová farmakologie   MeSH
• kyselina arachidonová antagonisté a inhibitory   MeSH
• lidé MeSH
• transformované buněčné linie MeSH
• umbeliferony farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• esculetin MeSH Prohlížeč
• indoly MeSH
• inhibitory cyklooxygenasy MeSH
• inhibitory lipoxygenas MeSH
• kyselina 5,8,11,14-eikosatetraenová MeSH
• kyselina arachidonová MeSH
• MK-886 MeSH Prohlížeč
• umbeliferony MeSH

INTRODUCTION: Cytochrome (CYP) epoxygenases (CYP2C and CYP2J) and soluble epoxide hydrolase (sEH) participate in the metabolism of arachidonic acid and may also have a potential role in enterocyte differentiation. The first critical step in the study of intestinal cell differentiation is the determination of a suitable in vitro model, which must be as similar as possible to the conditions of a living organism. It is known that HT-29 and Caco2 cell lines derived from human colorectal carcinomas can differentiate into enterocyte-like cells in appropriate culture conditions. MATERIAL AND METHODS: We tested 4 different approaches of enterocyte-like differentiation and determined the most appropriate culture conditions for each model. Subsequently, the changes in the expression of CYP epoxygenases and sEH in undifferentiated and differentiated cells were measured by In-Cell ELISA. These results were compared with immunohistochemical profiles of expression of CYP epoxygenases and sEH in samples of human embryonic and fetal intestines as well as adult duodenum and colon. RESULTS: Our results show that sodium butyrate (NaBt)-differentiated HT-29 cells and spontaneously differentiated Caco2 cells resemble CYP epoxygenases and sEH profiles, corresponding with different types of intestines. CONCLUSION: Our study revealed that the most suitable models for the study of the role of CYP epoxygenases and sEH expression in differentiation of intestinal epithelium are NaBt-differentiated HT-29 cells and spontaneously differentiated Caco2 cells.

• Klíčová slova
• Arachidonic acid metabolism, Differentiation, In vitro model, Intestine,
• MeSH
• buněčná diferenciace * MeSH
• buňky HT-29 MeSH
• Caco-2 buňky MeSH
• enterocyty enzymologie   MeSH
• epoxid hydrolasy metabolismus   MeSH
• kyselina arachidonová metabolismus   MeSH
• lidé MeSH
• střeva cytologie   embryologie   MeSH
• střevní sliznice * embryologie   metabolismus   MeSH
• systém (enzymů) cytochromů P-450 metabolismus   MeSH
• techniky in vitro MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• epoxid hydrolasy MeSH
• kyselina arachidonová MeSH
• systém (enzymů) cytochromů P-450 MeSH

The effect of arachidonic acid in 5.10(-4) and 5.10(-5) mol.l-1 concentration (as the Na salt, SIGMA) on ouabain-sensitive ATPase (E. C. 3.6.1.3) activity was studied in the cerebral cortex and medulla oblongata of 5-day-old and adult rats. In adult rats, arachidonic acid significantly inhibited ouabain-sensitive ATPase activity in both the cerebral cortex and the medulla oblongata. In 5-day-old rats, only the higher concentration (5.10(-4) mol.l-1) inhibited the enzyme statistically significantly; use of the lower concentration was not followed by any significant changes in Na+-K+-ATPase activity.

• MeSH
• inbrední kmeny potkanů MeSH
• krysa rodu Rattus MeSH
• kyselina arachidonová MeSH
• kyseliny arachidonové farmakologie   MeSH
• medulla oblongata enzymologie   MeSH
• mozková kůra enzymologie   MeSH
• sodíko-draslíková ATPasa antagonisté a inhibitory   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kyselina arachidonová MeSH
• kyseliny arachidonové MeSH
• sodíko-draslíková ATPasa MeSH

The dose-dependent inhibition of thrombin stimulated platelet aggregation due to beta-adrenoceptor blocking drugs followed the rank order of potency: propranolol greater than alprenolol greater than metipranolol and correlated with arachidonic acid (3H-AA) liberation. Atenolol which slightly potentiated stimulated aggregation increased also the liberation of 3H-AA from membrane phospholipids of isolated platelets. Stimulation of platelets resulted in decreased 3H-AA incorporation into phosphatidylcholine, phosphatidylinositol and phosphatidic acid and increased incorporation into phosphatidylethanolamine and phosphatidylserine. Alprenolol, metipranolol and propranolol enhanced the incorporation of 3H-AA into phosphatidylcholine of stimulated platelets.

• MeSH
• alprenolol farmakologie   MeSH
• atenolol farmakologie   MeSH
• beta blokátory farmakologie   MeSH
• fosfatidylcholiny metabolismus   MeSH
• fosfatidylethanolaminy metabolismus   MeSH
• fosfatidylinositoly metabolismus   MeSH
• hydrolýza MeSH
• inhibitory agregace trombocytů farmakologie   MeSH
• kyselina arachidonová MeSH
• kyseliny arachidonové metabolismus   MeSH
• metipranolol farmakologie   MeSH
• propranolol farmakologie   MeSH
• rozpustnost MeSH
• trombin fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• alprenolol MeSH
• atenolol MeSH
• beta blokátory MeSH
• fosfatidylcholiny MeSH
• fosfatidylethanolaminy MeSH
• fosfatidylinositoly MeSH
• inhibitory agregace trombocytů MeSH
• kyselina arachidonová MeSH
• kyseliny arachidonové MeSH
• metipranolol MeSH
• propranolol MeSH
• trombin MeSH

Propranolol (PRO), alprenolol (ALP), metipranolol (MET) and oxprenolol (OXP) inhibited arachidonic acid (AA) liberation from membrane phospholipids, malondialdehyde (MDA) formation and thromboxane B2 (TXB2) production in stimulated platelets. The inhibition was dose-dependent and with slight variations followed the rank order of potency: PRO greater than or equal to ALP greater than or equal to MET greater than or equal to OXP. Atenolol (ATE) was without any inhibitory effect. Inhibition of the arachidonic acid pathway in stimulated platelets may significantly contribute to the antiaggregatory effect of beta-adrenoceptor blocking drugs.

• MeSH
• beta blokátory farmakologie   MeSH
• inhibitory agregace trombocytů farmakologie   MeSH
• krysa rodu Rattus MeSH
• kyselina arachidonová krev   fyziologie   MeSH
• malondialdehyd krev   MeSH
• techniky in vitro MeSH
• trombin farmakologie   MeSH
• trombocyty účinky léků   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• beta blokátory MeSH
• inhibitory agregace trombocytů MeSH
• kyselina arachidonová MeSH
• malondialdehyd MeSH
• trombin MeSH

The Na+-dependent high-affinity choline uptake (HACU) transport and the [3H]hemicholinium-3 ([3H]HC-3) specific binding were measured on hippocampal synaptosomes of young (3-6 months) and old (22 months) Wistar rats. In vitro effects of 100-300 microM arachidonic acid (AA) and of 5% ethanol were tested under basal as well as stimulated (55 mM KCl) conditions. The influence of AA (an irreversible decrease of HACU and a reversible increase of [3H]HC-3 binding) was more marked under stimulated rather than basal conditions in brain tissue of young rats. The increased K+-depolarization effect on HACU and the decreased influence of AA on [3H]HC-3 binding were estimated in brain tissue of old compared to young rats. Results suggest the involvement of different pools of the high-affinity choline carrier and marked changes due to aging in the regulation of the HACU transport.

• MeSH
• biologický transport MeSH
• cholin metabolismus   MeSH
• draslík farmakologie   MeSH
• ethanol farmakologie   MeSH
• hemicholinium 3 metabolismus   MeSH
• hipokampus účinky léků   metabolismus   MeSH
• krysa rodu Rattus MeSH
• kyselina arachidonová farmakologie   MeSH
• potkani Wistar MeSH
• stárnutí metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cholin MeSH
• draslík MeSH
• ethanol MeSH
• hemicholinium 3 MeSH
• kyselina arachidonová MeSH

The lipophilic beta-adrenoceptor blocking (BAB) drugs metipranolol, propranolol and exaprolol significantly decreased 48/80- and A23187-induced 32P incorporation into rat mast cell phospholipids. Exaprolol was the most active, followed by propranolol and metipranolol. Atenolol and metipranolol significantly decreased the 48/80-stimulated, and metipranolol and exaprolol the A23187-stimulated 3H-arachidonic acid liberation from isolated mast cells.

• MeSH
• beta blokátory farmakologie   MeSH
• calcimycin farmakologie   MeSH
• fosfáty metabolismus   MeSH
• fosfolipidy metabolismus   MeSH
• inbrední kmeny potkanů MeSH
• krysa rodu Rattus MeSH
• kyselina arachidonová MeSH
• kyseliny arachidonové metabolismus   MeSH
• mastocyty účinky léků   metabolismus   MeSH
• p-methoxy-N-methylfenethylamin farmakologie   MeSH
• techniky in vitro MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• beta blokátory MeSH
• calcimycin MeSH
• fosfáty MeSH
• fosfolipidy MeSH
• kyselina arachidonová MeSH
• kyseliny arachidonové MeSH
• p-methoxy-N-methylfenethylamin MeSH