Hyaluronan (HA) effects on immune response are suggested to be dependent on HA molecular weight (MW), as low MW HA should activate immune cells in contrast to high MW HA. However, some current studies do not support this conception and emphasize the importance of the form of preparation of HA, particularly with respect to its purity and origin. We compared the activation of mouse immune cells by HA samples (100kDa, 500kDa, and 997kDa) prepared from HA originating from rooster comb, and HA samples (71kDa, 500kDa, and 1000kDa) prepared from pharmacological grade HA originating from Streptococcus equi. Interestingly, in contrast to established theory, only middle and high MW HA originating from rooster comb induced the production of tumor necrosis factor-α by macrophages and in whole blood. Further, all tested preparations of HA failed to induce the expression of inducible nitric oxide synthase, the production of nitric oxide, or the expression of cyclooxygenase 2 in macrophages and splenocytes. Importantly, all HA samples originating from rooster comb were found to be contaminated by endotoxin (up to 1.23EU/ml). Hence, low MW HA did not reveal itself to have significantly higher immunostimulatory activity compared to HA of higher MW.
- MeSH
- buněčná imunita účinky léků genetika MeSH
- cyklooxygenasa 2 genetika MeSH
- kur domácí MeSH
- kyselina hyaluronová farmakologie MeSH
- makrofágy účinky léků imunologie MeSH
- molekulová hmotnost MeSH
- myši MeSH
- oxid dusnatý genetika MeSH
- RAW 264.7 buňky MeSH
- regulace genové exprese účinky léků MeSH
- Streptococcus equi chemie MeSH
- synthasa oxidu dusnatého genetika MeSH
- TNF-alfa genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Activation of nuclear factor-κB (NF-κB) by increased production of reactive oxygen species (ROS) might induce transcription and expression of different antioxidant enzymes and also of nitric oxide synthase (NOS) isoforms. Thus, we aimed at studying the effect of NF-κB inhibition, caused by JSH-23 (4-methyl-N (1)-(3-phenyl-propyl)-benzene-1,2-diamine) injection, on ROS and NO generation in hereditary hypertriglyceridemic (HTG) rats. 12-week-old, male Wistar and HTG rats were treated with JSH-23 (bolus, 10 μmol, i.v.). After one week, blood pressure (BP), superoxide dismutase (SOD) activity, SOD1, endothelial NOS (eNOS), and NF-κB (p65) protein expressions were higher in the heart of HTG rats compared to control rats. On the other hand, NOS activity was decreased. In HTG rats, JSH-23 treatment increased BP and heart conjugated dienes (CD) concentration (measured as the marker of tissue oxidative damage). Concomitantly, SOD activity together with SOD1 expression was decreased, while NOS activity and eNOS protein expression were increased significantly. In conclusion, NF-κB inhibition in HTG rats led to decreased ROS degradation by SOD followed by increased oxidative damage in the heart and BP elevation. In these conditions, increased NO generation may represent rather a counterregulatory mechanism activated by ROS. Nevertheless, this mechanism was not sufficient enough to compensate BP increase in HTG rats.
- MeSH
- exprese genu účinky léků MeSH
- fenylendiaminy farmakologie MeSH
- glutathion analýza MeSH
- hyperlipoproteinemie typ IV patologie veterinární MeSH
- krevní tlak účinky léků MeSH
- krysa rodu rattus MeSH
- myokard metabolismus MeSH
- oxid dusnatý metabolismus MeSH
- oxidační stres účinky léků MeSH
- potkani Wistar MeSH
- reaktivní formy kyslíku metabolismus MeSH
- srdeční komory metabolismus MeSH
- superoxiddismutasa genetika metabolismus MeSH
- synthasa oxidu dusnatého, typ III genetika metabolismus MeSH
- synthasa oxidu dusnatého genetika metabolismus MeSH
- tělesná hmotnost účinky léků MeSH
- transkripční faktor RelA genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
It is known that cells and organisms can indirectly "sense" changes in L-arginine availability via changes in the activity of various metabolic pathways. However, the mechanism(s) by which genes can be directly regulated by L-arginine in mammalian cells have not yet been elucidated. We investigated the effect of L-arginine in the in vivo model of peritoneal inflammation in mice and in vitro in RAW 264.7 macrophages. A detailed analysis of basic physiological functions and selected intracellular signaling cascades revealed that L-arginine is crucial for the acceleration of macrophage activation by bacterial lipopolysaccharide. L-arginine increased the production of reactive oxygen species, nitric oxide, release of Ca(2+), as well as inducible nitric oxide synthase expression. Interestingly, the effect of L-arginine on macrophage activation was dependent on the phosphorylation of mitogen-activated protein kinases and activity of phospholipase C. In RAW 264.7 cells, L-arginine was shown to modulate the response of macrophages toward lipopolysaccharide via the activation of G-protein-coupled receptors. According to our data, we concluded that L-arginine availability plays a key role in the initiation of intracellular signaling pathways that trigger the lipopolysaccharide-induced inflammatory responses in murine macrophages. Although macrophages are partially stimulated in the absence of extracellular L-arginine, the presence of this amino acid significantly accelerates the sensitivity of macrophages to bacterial endotoxin.
- MeSH
- arginin imunologie farmakologie MeSH
- buněčné linie MeSH
- dieta MeSH
- endotoxiny imunologie MeSH
- extracelulárním signálem regulované MAP kinasy metabolismus MeSH
- fosfolipasy typu C metabolismus MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- oxid dusnatý metabolismus MeSH
- peritoneální makrofágy účinky léků imunologie MeSH
- peritonitida imunologie MeSH
- signální transdukce účinky léků imunologie MeSH
- synthasa oxidu dusnatého genetika metabolismus MeSH
- techniky in vitro MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Both brain and peripheral nitric oxide (NO) play a role in the control of blood pressure and circulatory homeostasis. Central NO production seems to counteract angiotensin II-induced enhancement of sympathetic tone. The aim of our study was to evaluate NO synthase (NOS) activity and protein expression of its three isoforms – neuronal (nNOS), endothelial NOS (eNOS) and inducible (iNOS) – in two brain regions involved in blood pressure control (diencephalon and brainstem) as well as in the kidney of young adult rats with either genetic (12-week-old SHR) or saltinduced hypertension (8-week-old Dahl rats). We have demonstrated reduced nNOS and iNOS expression in brainstem of both hypertensive models. In SHR this abnormality was accompanied by attenuated NOS activity and was corrected by chronic captopril treatment which prevented the development of genetic hypertension. In salt hypertensive Dahl rats nNOS and iNOS expression was also decreased in the diencephalon where neural structures important for salt hypertension development are located. As far as peripheral NOS activity and expression is concerned, renal eNOS expression was considerably reduced in both genetic and salt-induced hypertension. In conclusions, we disclosed similar changes of NO system in the brainstem (but not in the diencephalon) of rats with genetic and salt-induced hypertension. Decreased nNOS expression was associated with increased blood pressure due to enhanced sympathetic tone.
- MeSH
- financování organizované MeSH
- hypertenze enzymologie etiologie genetika MeSH
- izoenzymy genetika metabolismus MeSH
- krysa rodu rattus MeSH
- kuchyňská sůl toxicita MeSH
- ledviny enzymologie metabolismus MeSH
- mozek enzymologie metabolismus MeSH
- oxid dusnatý metabolismus MeSH
- potkani inbrední Dahl MeSH
- synthasa oxidu dusnatého genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
In this paper we assessed: (i) the change in nitric oxide synthase (NOS) isoforms' expression and intracellular localization and in NOS mRNA in porcine oocytes during meiotic maturation; (ii) the effect of NOS inhibition by N(omega)-nitro-l-arginine methyl ester (l-NAME) and aminoguanidine (AG) on meiotic maturation of cumulus-oocyte complexes (COC) as well as denuded oocytes (DO); and (iii) nitric oxide (NO) formation in COC. All three NOS isoforms (eNOS, iNOS and nNOS) and NOS mRNA (eNOS mRNA, iNOS mRNA and nNOS mRNA) were found in both porcine oocytes and their cumulus cells except for nNOS mRNA, which was not detected in the cumulus cells. NOS isoforms differed in their intracellular localization in the oocyte: while iNOS protein was dispersed in the oocyte cytoplasm, nNOS was localized in the oocyte cytoplasm and in germinal vesicles (GV) and eNOS was present in dots in the cytoplasm, GV and was associated with meiotic spindles. l-NAME inhibitor significantly suppressed metaphase (M)I to MII transition (5.0 mM experimental group: 34.9% MI, control group: 9.5% MI) and at the highest concentration (10.0 mM) also affected GV breakdown (GVBD); in contrast also AG inhibited primarily GVBD. The majority of the oocytes (10.0 mM experimental group: 60.8%, control group: 1.2%) was not able to resume meiosis. AG significantly inhibited GVBD in DO, but l-NAME had no significant effect on the GVBD of these cells. During meiotic maturation, NO is formed in COC and the NO formed by cumulus cells is necessary for the process of GVBD.
- MeSH
- konfokální mikroskopie MeSH
- kumulární buňky enzymologie MeSH
- meióza MeSH
- messenger RNA metabolismus MeSH
- NG-nitroargininmethylester farmakologie MeSH
- oocyty účinky léků enzymologie MeSH
- protein - isoformy antagonisté a inhibitory genetika metabolismus MeSH
- synthasa oxidu dusnatého antagonisté a inhibitory genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Koronární nemoc srdecního štepu (KNSŠ) predstavuje jednu z nejzávažnejších komplikací transplantace srdce (TS). Za základní patofyziologický moment v rozvoji KNSŠ je považována endoteliální dysfunkce, zpusobená množstvím faktoru imunologické i neimunologické povahy. Klícovou úlohu v regulaci rady procesu na úrovni cévní steny hraje oxid dusnatý (NO), produkovaný endoteliální syntázou oxidu dusnatého (eNOS). Polymorfismy genu pro eNOS, ovlivnující dostupnost a aktivitu tohoto enzymu, mohou ovlivnovat dostupnost NO na úrovni cévní steny, a vzhledem k významným ochranným vlastnostem NO i náchylnost vencitých tepen srdecního štepu k rozvoji KNSŠ. Cíl: Cílem práce je posoudit vliv genových polymorfismu pro eNOS (intron 4, T786C, G894T) na rozvoj morfologických známek a výskyt klinických manifestací (výskyt akutního koronárního syndromu, maligní poruchy rytmu, náhlé smrti, srdecního selhání pri vyloucení ostatních prícin) KNSŠ u príjemcu TS. Studovaná populace a metody: Studovaná populace byla tvorena príjemci TS, u nichž bylo casne v jednom roce po TS provedeno intra vaskulární ultrazvukové vyšetrení vencitých tepen srdecního štepu. Soucasne byla u jejich korespondujících dárcu provedena genotypizace eNOS ve výše uvedených oblastech. Výsledky: Byl nalezen statisticky významný vztah mezi alelou 4b a genotypem 4b4b (intron 4) na strane jedné a výskytem jak casne, tak v jednom roce po TS detekovaných morfologických známek KNSŠ na strane druhé. Alela 4b a genotyp 4b4b byly shledány nezávislými rizikovými faktory pro prítomnost morfologických známek KNSŠ casne po TS. Dále byl nalezen statisticky významný vztah mezi alelou C a genotypem CC (T786C) a výskytem klinických manifestací KNSŠ. Záver: Byla nalezena pozitivní korelace mezi genovým polymorfismem pro eNOS v oblasti intron 4 a prítomností morfologických známek KNSŠ jak casne, tak v jednom roce po TS. Genotyp 4b4b a alela 4b intron-4 genového polymorfismu pro eNOS byly shledány rizikovými faktory pro prítomnost morfologických známek casne detekované KNSŠ. Rovnež tak byla nalezena pozitivní korelace mezi genovým polymorfismem T785C pro eNOS a výskytem klinických manifestací KNSŠ.
Introduction: Cardiac graft coronary artery disease (CGCAD) constitutes of the most serious complications of heart transplantation (HTx). The principal pathophysiological trigger in the development of CGCAD is believed to be endothelial dysfunction due to a number of immunological and non-immunological factors. The key role in the regulation of a variety of processes at the level of the vessel wall is played by nitric oxide (NO), produced by endothelial nitric oxide synthase (eNOS). eNOS gene polymorphisms, a/ecting the availability and activity of this enzyme, may modulate NO availability at the level of the vessel wall and, given the significant protective properties of NO, also the susceptibility of the coronary arteries of the cardiac graft to develop CGCAD. Aim: The aim of the study was to assess the e/ect of eNOS gene polymorphisms (intron 4, T786C, G894T) on the development of morphological traits and occurrence of clinical presentations (incidence of acute coronary syndrome, malignant arrhythmias, sudden death, heart failure after exclusion of other causes) of CGCAD in HTx recipients. Study population and methods: The study population was made up by HTx recipients having intravascular ultrasound of the cardiac graft coronary vessels early and at 1 year post-HTx. At the same time, matched donors had eNOS genotyping in the above regions. Results: A signifcant correlation was found between the 4b allele and 4b4b genotype (intron 4) on the one hand and the incidence of morphological traits of CGCAD, detected both early and at 1 year post-HTx on the other. The 4b allele and the 4b4b genotype were identified as independent risk factors for the presence of morphological traits of CGCAD early post-HTx. A significant correlation was also demonstrated between the C allele and the CC genotype (T786C) on the one hand, and the incidence of clinical presentations of CGCAD on the other. Conclusion: A correlation was found between the eNOS gene polymorphism in the region of intron 4 and the presence of morphological traits of CGCAD both early and at 1 year post-HTx. The 4b4b genotype and the 4b allele of the eNOS gene polymorphism were identified as risk factors for the presence of morphological traits of CGCAD detected early. Likewise, a correlation was found between the eNOS T785C gene polymorphism and the incidence of clinical presentations of CGCAD.
- MeSH
- akutní koronární syndrom etiologie genetika MeSH
- dospělí MeSH
- financování organizované MeSH
- koronární okluze etiologie genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- oxid dusnatý nedostatek MeSH
- polymorfismus genetický MeSH
- synthasa oxidu dusnatého genetika MeSH
- transplantace srdce škodlivé účinky MeSH
- vzorkové studie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
Certain liver metabolic diseases point to the presence of disturbances in glycogen deposition. Epinephrine raises the cAMP level that activates protein kinase A leading to the activation of phosphorylase and glycogen breakdown. In the present report, we sought to investigate whether NO is produced during adrenoceptor agonist-induced glycogenolysis in rat hepatocytes in cultures. Isolated glycogen rich rat hepatocytes in cultures were used. NO production (NO2 -) was assessed under the effect of adrenergic agonists and adrenergic agonist/antagonist pairs, dibutyryl cyclic AMP sodiumpotassium salt (db-cAMP), NO synthase (NOS) inhibitors N?-nitro-L-arginine methyl ester (L-NAME), aminoguanidine (AG) and the NO donor S-nitroso-N-acetyl penicillamine (SNAP). The inducible NO synthase (iNOS) mRNA was examined by the reverse transcription-polymerase chain reaction (RT-PCR). Glycogenolysis was quantified by glucose levels released into medium. The amount of glucose and NO2 - released by hepatocytes was increased as a result of epinephrine, phenylephrine or db-cAMP treatments. The increase in glucose and NO2 - released by epinephrine or phenylephrine was blocked or reduced by prazosin pretreatment and by NOS inhibitors aminoguanidine and LNAME. iNOS gene expression was up-regulated by epinephrine. It can be concluded that glycogenolysis occurs through ?-adrenoceptor stimulation and a signaling cascade may involve NO production.
- MeSH
- adrenalin genetika metabolismus MeSH
- financování organizované využití MeSH
- glykogenolýza genetika účinky léků MeSH
- hepatocyty metabolismus účinky léků MeSH
- jaterní glykogen chemie metabolismus MeSH
- nemoci jater etiologie komplikace metabolismus MeSH
- oxid dusnatý metabolismus MeSH
- polymerázová řetězová reakce s reverzní transkripcí metody využití MeSH
- potkani Wistar metabolismus MeSH
- synthasa oxidu dusnatého genetika metabolismus MeSH
- MeSH
- antigeny CD4 imunologie MeSH
- antigeny CD8 imunologie MeSH
- cytokininy biosyntéza MeSH
- finanční podpora výzkumu jako téma MeSH
- lidé MeSH
- monoklonální protilátky farmakologie MeSH
- myši inbrední BALB C MeSH
- oxid dusnatý biosyntéza MeSH
- potkani inbrední LEW MeSH
- synthasa oxidu dusnatého genetika metabolismus MeSH
- transplantace heterologní MeSH
- transplantace rohovky MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- MeSH
- aorta patologie MeSH
- arterioskleróza diagnóza MeSH
- cévní endotel patologie MeSH
- finanční podpora výzkumu jako téma MeSH
- homocystein analýza krev MeSH
- lidé MeSH
- synthasa oxidu dusnatého genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- kongresy MeSH
