N-acetylcysteine (NAC), often used as an antioxidant-scavenging reactive oxygen species (ROS) in vitro, was recently shown to increase the cytotoxicity of other compounds through ROS-dependent and ROS-independent mechanisms. In this study, NAC itself was found to induce extensive ROS production in human leukemia HL-60 and U937 cells. The cytotoxicity depends on ROS-modulating enzyme expression. In HL-60 cells, NAC activated NOX2 to produce superoxide (O2•-). Its subsequent conversion into H2O2 by superoxide dismutase 1 and 3 (SOD1, SOD3) and production of ClO- from H2O2 by myeloperoxidase (MPO) was necessary for cell death induction. While the addition of extracellular SOD potentiated NAC-induced cell death, extracellular catalase (CAT) prevented cell death in HL-60 cells. The MPO inhibitor partially reduced the number of dying HL-60 cells. In U937 cells, the weak cytotoxicity of NAC is probably caused by lower expression of NOX2, SOD1, SOD3, and by the absence of MOP expression. However, even here, the addition of extracellular SOD induced cell death in U937 cells, and this effect could be reversed by extracellular CAT. NAC-induced cell death exhibited predominantly apoptotic features in both cell lines. Conclusions: NAC itself can induce extensive production of O2•- in HL-60 and U937 cell lines. The fate of the cells then depends on the expression of enzymes that control the formation and conversion of ROS: NOX, SOD, and MPO. The mode of cell death in response to NAC treatment bears apoptotic and apoptotic-like features in both cell lines.

• Klíčová slova
• HL-60 cells, MPO, N-acetylcysteine, NOX, SOD, U937 cells, oxidative stress,
• MeSH
• acetylcystein farmakologie   MeSH
• HL-60 buňky MeSH
• katalasa genetika   MeSH
• leukemie farmakoterapie   genetika   metabolismus   MeSH
• lidé MeSH
• NADPH-oxidasa 2 genetika   MeSH
• oxidační stres účinky léků   MeSH
• peroxidasa genetika   MeSH
• proliferace buněk účinky léků   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• stanovení celkové genové exprese MeSH
• superoxiddismutasa genetika   MeSH
• U937 buňky MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• acetylcystein MeSH
• CYBB protein, human MeSH Prohlížeč
• katalasa MeSH
• MPO protein, human MeSH Prohlížeč
• NADPH-oxidasa 2 MeSH
• peroxidasa MeSH
• reaktivní formy kyslíku MeSH
• superoxiddismutasa MeSH

Autoinflammatory diseases are characterized by dysregulation of the innate immune system, leading to spontaneous inflammation. Pstpip2cmo mouse strain is a well-characterized model of this class of disorders. Because of the mutation leading to the lack of adaptor protein PSTPIP2, these animals suffer from autoinflammatory chronic multifocal osteomyelitis similar to several human syndromes. Current evidence suggests that it is driven by hyperproduction of IL-1β by neutrophil granulocytes. In this study, we show that in addition to IL-1β, PSTPIP2 also negatively regulates pathways governing reactive oxygen species generation by neutrophil NOX2 NADPH oxidase. Pstpip2cmo neutrophils display highly elevated superoxide production in response to a range of stimuli. Inactivation of NOX2 NADPH oxidase in Pstpip2cmo mice did not affect IL-1β levels, and the autoinflammatory process was initiated with similar kinetics. However, the bone destruction was almost completely alleviated, suggesting that dysregulated NADPH oxidase activity is a key factor promoting autoinflammatory bone damage in Pstpip2cmo mice.

• MeSH
• adaptorové proteiny signální transdukční genetika   metabolismus   MeSH
• buněčné linie MeSH
• cytoskeletální proteiny genetika   metabolismus   MeSH
• interleukin-1beta imunologie   metabolismus   MeSH
• kosti a kostní tkáň imunologie   patologie   MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• mutace MeSH
• myši transgenní MeSH
• myši MeSH
• NADPH-oxidasa 2 genetika   metabolismus   MeSH
• neutrofily imunologie   metabolismus   MeSH
• osteomyelitida genetika   imunologie   patologie   MeSH
• primární buněčná kultura MeSH
• signální transdukce genetika   imunologie   MeSH
• superoxidy imunologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• Cybb protein, mouse MeSH Prohlížeč
• cytoskeletální proteiny MeSH
• IL1B protein, mouse MeSH Prohlížeč
• interleukin-1beta MeSH
• NADPH-oxidasa 2 MeSH
• Pstpip2 protein, mouse MeSH Prohlížeč
• superoxidy MeSH

P38alpha kinase plays an important role in the regulation of both cell stress response and cell fate. In this study, we report that p38alpha kinase-deficient embryonic stem cells exhibit a higher production of reactive oxygen species (ROS) in contrast to their wild-type counterpart. Analysis of the expressions of NADPH oxidases (NOXs) and dual oxidases, crucial enzymes involved in intracellular ROS formation, shows NOX2/gp91phox is over-expressed in p38alpha deficient cells. The particular increase in superoxide formation was confirmed by the specific detection of hydroethidine derivate 2-hydroxyethidium. ROS formation decreased when the level of NOX2 was silenced by siRNA in p38alpha deficient cells. These data suggest the importance of p38alpha kinase in the regulation of ROS metabolism in embryonic stem cells and the significance of the observed phenomena of cancer cell-like phenotypes, which is discussed.

• Klíčová slova
• Embryonic stem cell, NADPH oxidase, Reactive oxygen species, p38 kinase,
• MeSH
• buněčná diferenciace fyziologie   MeSH
• genový knockdown MeSH
• genový knockout MeSH
• kultivované buňky MeSH
• membránový potenciál mitochondrií fyziologie   MeSH
• mitochondrie metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 14 genetika   metabolismus   MeSH
• myší embryonální kmenové buňky metabolismus   MeSH
• myši MeSH
• NADPH-oxidasa 2 genetika   metabolismus   MeSH
• superoxidy metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Cybb protein, mouse MeSH Prohlížeč
• mitogenem aktivovaná proteinkinasa 14 MeSH
• NADPH-oxidasa 2 MeSH
• superoxidy MeSH

BACKGROUND AND AIM: Increased homocysteine (Hcy) is associated with coronary artery disease (CAD). Hcy increases reactive oxygen species (ROS) via NADPH oxidases (Nox), reducing acetylcholine-mediated vasorelaxation. We aimed to determine if putative Nox2 inhibitors prevent Hcy-impaired acetylcholine-mediated vasorelaxation. METHODS AND RESULTS: New Zealand White rabbit and wild-type (C57BL/6) and Nox2-/- (NOX) mice aortic rings were mounted in organ baths. Rabbit rings were incubated with either apocynin (10 μM), gp91ds-tat (GP, 1 μM) or PhoxI2 (1 μM) and mice rings GP (1 μM) only. Some rabbit rings were incubated with 3 mM Hcy, before pre-contraction, followed by dose-response relaxation to acetylcholine (ACh; 0.01μM-10μM). In rabbit rings treated with Hcy and GP, O2‾ donor pyrogallol (1 μM) or Akt activator SC79 (1 μM) was added 5 min before ACh. Mice rings were used to compare Nox2 deletion to normal acetylcholine-mediated relaxation. In rabbits, Hcy reduced acetylcholine-mediated relaxation vs. control (p < 0.0001). Treatment + Hcy reduced relaxation compared with treatment alone (p < 0.0001). Pyrogallol and SC79 reversed the response of GP + Hcy (p = 0.0001). In mice, Nox2 deletion reduced acetylcholine-mediated vasorelaxation. Rabbit tissue analysis revealed that Hcy reduced eNOS phosphorylation at Thr495 and increased eNOS phosphorylation at Ser1177; no further alteration at Thr495 was observed with GP. In contrast, GP prevented increased phosphorylation at Ser1177. CONCLUSIONS: Apocynin, GP and PhoxI2 worsens acetylcholine-mediated vascular relaxation in rabbit aorta, which is supported by results from mouse Nox2 deletion data. These inhibitors worsen Hcy-induced vascular dysfunction, suggesting that current putative Nox2 inhibitors might not be useful in treating HHcy.

• Klíčová slova
• Acetylcholine, Acetylcholine-mediated vasorelaxation, Homocysteine, Nox2, Pharmacology,
• MeSH
• acetofenony farmakologie   MeSH
• acetylcholin farmakologie   MeSH
• aorta účinky léků   enzymologie   MeSH
• fosforylace MeSH
• glykoproteiny farmakologie   MeSH
• homocystein farmakologie   MeSH
• inhibitory enzymů farmakologie   MeSH
• králíci MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• NADPH-oxidasa 2 antagonisté a inhibitory   genetika   metabolismus   MeSH
• serin MeSH
• synthasa oxidu dusnatého, typ III metabolismus   MeSH
• techniky in vitro MeSH
• threonin MeSH
• vazodilatace účinky léků   MeSH
• vazodilatancia farmakologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• acetofenony MeSH
• acetovanillone MeSH Prohlížeč
• acetylcholin MeSH
• Cybb protein, mouse MeSH Prohlížeč
• glykoproteiny MeSH
• gp91ds-tat protein, chimeric MeSH Prohlížeč
• homocystein MeSH
• inhibitory enzymů MeSH
• NADPH-oxidasa 2 MeSH
• Nos3 protein, mouse MeSH Prohlížeč
• serin MeSH
• synthasa oxidu dusnatého, typ III MeSH
• threonin MeSH
• vazodilatancia MeSH

This short review article summarizes what is known clinically and biochemically about the seven human NADPH oxidases. Emphasis is put on the connection between mutations in the catalytic and regulatory subunits of Nox2, the phagocyte defense enzyme, with syndromes like chronic granulomatous disease, as well as a number of chronic inflammatory diseases. These arise paradoxically from a lack of reactive oxygen species production needed as second messengers for immune regulation. Both Nox2 and the six other human NADPH oxidases display signaling functions in addition to the functions of these enzymes in specialized biochemical reactions, for instance, synthesis of the hormone thyroxine. NADPH oxidases are also needed by Saccharomyces cerevisiae cells for the regulation of the actin cytoskeleton in times of stress or developmental changes, such as pseudohyphae formation. The article shows that in certain cancer cells Nox4 is also involved in the re-structuring of the actin cytoskeleton, which is required for cell mobility and therefore for metastasis.

In diesem kurzen Übersichtsbeitrag fassen wir den klinischen und biochemischen Wissensstand zu den sieben menschlichen NADPH-Oxidasen zusammen. Ein Schwerpunkt liegt dabei auf dem Zusammenhang zwischen Mutationen in den katalytischen und regulatorischen Untereinheiten von Nox2 – der NADPH-Oxidase der Phagozyten mit Abwehrfunktion – und Syndromen wie der septischen Granulomatose, aber auch einer Reihe von chronisch-entzündlichen Erkrankungen. Hervorgerufen werden diese durch einen Mangel an reaktiven Sauerstoffspezies, welche als Produkte der NADPH-Oxidase-Reaktion für die Immunregulation notwendig sind. Sowohl Nox2 als auch die anderen sechs menschlichen NADPH-Oxidasen haben neben ihren Funktionen in spezialisierten biochemischen Reaktionen (Beispiel: Thyroxinsynthese) auch eine Funktion in der Signaltransduktion. In der Hefe Saccharomyces cerevisiae wird die NADPH-Oxidase Yno1 als Signalgeber für die Restrukturierung des Aktinzytoskeletts gebraucht, z. B. unter Stress oder bei der Bildung von Pseudohyphen. In bestimmten Tumorzellen ist die NADPH-Oxidase Nox4 ebenfalls an der Restrukturierung des Aktinzytoskeletts beteiligt – eine Voraussetzung für die Zellmobilität und damit auch für die Metastasierung dieser Tumoren.

• Klíčová slova
• Actin cytoskeleton, Chronic granulomatous disease, Nox enzymes, Reactive oxygen species, Saccharomyces cerevisiae,
• MeSH
• eukaryotické buňky * MeSH
• lidé MeSH
• NADPH-oxidasa 2 MeSH
• NADPH-oxidasa 4 MeSH
• NADPH-oxidasy * fyziologie   MeSH
• reaktivní formy kyslíku MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• CYBB protein, human MeSH Prohlížeč
• NADPH-oxidasa 2 MeSH
• NADPH-oxidasa 4 MeSH
• NADPH-oxidasy * MeSH
• NOX4 protein, human MeSH Prohlížeč
• reaktivní formy kyslíku MeSH

UNLABELLED: Introduction and aim. Hepatocellular carcinoma (HCC) is the most common primary malignant liver tumor. It is primarily caused by hepatic cirrhosis or chronic viral hepatitis. Hepatic carcinogenesis is associated with increased oxidative stress. Thus, the aim of our study was to assess expression of the genes involved in the homeostasis of oxidative stress in patients with HCC. MATERIAL AND METHODS: The study was performed on 32 patients with primary HCC (verified by liver histology in 29 patients) and 27 control subjects (in 11 subjects, liver histology was available either with no or minimal changes in the liver tissue). Gene expressions of heme oxygenase 1 (HMOX1), biliverdin reductase A/B (BLVRA/B), NADPH oxidase 2 (NOX2) and p22phox were analyzed in the liver and peripheral blood leukocytes (PBL) in the subjects. RESULTS: Compared to controls, almost a 3 times higher mRNA level of BLVRA was detected in livers of HCC patients (p = 0.002); while those of BLVRB as well as HMOX1 were unchanged (p > 0.05). In accord with these results in the liver tissue, BLVRA mRNA levels in PBL were also significantly increased in HCC patients (p = 0.012). mRNA levels of NOX2 and p22phox in the liver tissue, although higher in HCC patients, did not differ significantly compared to control subjects (p > 0.05). Nevertheless, NOX2 mRNA level in PBL was significantly higher in HCC patients (p = 0.003). CONCLUSIONS: BLVRA mRNA levels in the liver as well as in PBL are significantly higher in HCC patients most likely as a feedback mechanism to control increased oxidative stress associated with HCC progression.

• MeSH
• hemoxygenasa-1 genetika   MeSH
• hepatocelulární karcinom krev   enzymologie   genetika   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové glykoproteiny genetika   MeSH
• messenger RNA genetika   MeSH
• nádorové biomarkery krev   genetika   MeSH
• nádory jater krev   enzymologie   genetika   patologie   MeSH
• NADPH-oxidasa 2 MeSH
• NADPH-oxidasy genetika   MeSH
• oxidační stres genetika   MeSH
• oxidoreduktasy působící na CH-CH vazby krev   genetika   MeSH
• progrese nemoci MeSH
• regulace genové exprese enzymů MeSH
• regulace genové exprese u nádorů MeSH
• senioři MeSH
• signální transdukce MeSH
• studie případů a kontrol MeSH
• upregulace MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biliverdin reductase MeSH Prohlížeč
• CYBA protein, human MeSH Prohlížeč
• CYBB protein, human MeSH Prohlížeč
• hemoxygenasa-1 MeSH
• HMOX1 protein, human MeSH Prohlížeč
• membránové glykoproteiny MeSH
• messenger RNA MeSH
• nádorové biomarkery MeSH
• NADPH-oxidasa 2 MeSH
• NADPH-oxidasy MeSH
• oxidoreduktasy působící na CH-CH vazby MeSH

INTRODUCTION: Nicotinamide adenine dinucleotide phosphate oxidases (NOX 1-5) are enzymes that generate cellular reactive oxygen species (ROS) besides mitochondria and might be important ROS sources associated with pregnancy complications, particularly preterm premature rupture of membranes (pPROM), that has been related to ROS. OBJECTIVE: To characterize NOX enzymes expression in human fetal membranes. METHODS: Differential expression and localization of NOX isoforms in human fetal membranes collected from women with uncomplicated pregnancies at term, preterm birth (PTB) or pPROM and in vitro in normal term membranes maintained in an organ explant system stimulated with water-soluble cigarette smoke extract (wsCSE) were documented by real time PCR and immunohistochemistry. RESULTS: Fetal membranes from term deliveries, PTB and pPROM expressed NOX 2, 3 and 4 mRNAs whereas NOX 1 and 5 were not detected. NOX 2 expression was 2.3-fold higher in PTB than pPROM (p = 0.005) whereas NOX 3 was 2.2-fold higher in pPROM compared to PTB (p = 0.04). NOX 2 and 3 expressions at term mimicked pPROM and PTB, respectively. No difference in NOX 4 expression was observed among the studied groups. NOX 2, 3 and 4 were localized to both amniotic and chorionic cells. Expression of NOX 2, 3 and 4 were not significant in wsCSE-stimulated membranes compared to untreated controls. DISCUSSION/CONCLUSIONS: NOX enzymes are present in the fetal membranes and are differentially expressed in PTB and pPROM. Absence of any changes in NOXs expression after wsCSE stimulation suggests ROS generation in the membranes does not always correlate with NOX expression.

• Klíčová slova
• Cigarette smoking extract, Oxidase enzymes, Preterm birth, Preterm premature rupture of membranes, Reactive species of oxygen,
• MeSH
• dospělí MeSH
• extraembryonální obaly enzymologie   MeSH
• kouření patofyziologie   MeSH
• lidé MeSH
• membránové glykoproteiny biosyntéza   MeSH
• membránové proteiny biosyntéza   MeSH
• NADPH-oxidasa 2 MeSH
• NADPH-oxidasy biosyntéza   MeSH
• novorozenec MeSH
• předčasný odtok plodové vody enzymologie   MeSH
• předčasný porod enzymologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• těhotenství MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CYBB protein, human MeSH Prohlížeč
• membránové glykoproteiny MeSH
• membránové proteiny MeSH
• NADPH-oxidasa 2 MeSH
• NADPH-oxidasy MeSH
• Nox3 protein, human MeSH Prohlížeč
• reaktivní formy kyslíku MeSH

Hyper-immunoglobulin (Ig)E syndrome (HIES) is a primary immunodeficiency associated with mutations in STAT3 resulting in impaired development of T helper type 17 (Th17) lymphocytes. HIES patients with a reduced frequency of Th17 cells present with infections caused by Staphylococcus aureus and/or Candida strains. The same spectrum of pathogens is present in patients with chronic granulomatous disease (CGD).We analysed the characteristics of the Th17 compartment in HIES and CGD. HIES patients showed very low numbers of Th17 cells. By contrast, the frequency of Th17 cells and production of Th17-derived cytokines was significantly higher among CGD patients when compared to both control samples and HIES. Naive CD4(+) cells in CGD patients had a normal capacity to differentiate into IL-17-producing cells and the numbers of Th17 cells in the CGD patients normalized following successful bone marrow transplantation. Our findings complement recent data on the importance of Th17 cells for elimination of infections with C. albicans and S. aureus.

• MeSH
• buněčná diferenciace imunologie   MeSH
• buňky Th17 imunologie   metabolismus   patologie   MeSH
• Candida albicans růst a vývoj   MeSH
• chronická granulomatózní nemoc genetika   imunologie   metabolismus   patologie   terapie   MeSH
• dítě MeSH
• dospělí MeSH
• ELISA MeSH
• homologní transplantace MeSH
• interferon gama imunologie   metabolismus   MeSH
• interleukin-17 imunologie   metabolismus   MeSH
• Jobův syndrom genetika   imunologie   metabolismus   patologie   MeSH
• kandidóza imunologie   mikrobiologie   MeSH
• lidé MeSH
• membránové glykoproteiny genetika   imunologie   metabolismus   MeSH
• mutace MeSH
• NADPH-oxidasa 2 MeSH
• NADPH-oxidasy genetika   imunologie   metabolismus   MeSH
• počet lymfocytů MeSH
• předškolní dítě MeSH
• průtoková cytometrie MeSH
• stafylokokové infekce imunologie   mikrobiologie   MeSH
• Staphylococcus aureus růst a vývoj   MeSH
• studie případů a kontrol MeSH
• transkripční faktor STAT3 genetika   imunologie   metabolismus   MeSH
• transplantace hematopoetických kmenových buněk MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CYBB protein, human MeSH Prohlížeč
• interferon gama MeSH
• interleukin-17 MeSH
• membránové glykoproteiny MeSH
• NADPH-oxidasa 2 MeSH
• NADPH-oxidasy MeSH
• transkripční faktor STAT3 MeSH