F.62. Essential Role of Nuclear Factor-kappa B for NADPH Oxidase Activity, CYBB and NCF-1 Gene Expression in Normal and Anhidrotic Ectodermal Dysplasia Leukocytes

2008 ◽  
Vol 127 ◽  
pp. S63
Author(s):  
Marcos Luengo ◽  
Carolina Prando ◽  
Jacinta Bustamante ◽  
Paulo-Vitor Pereira ◽  
Walmir Aragão-Filho ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nadph Oxidase ◽  
Oxidase Activity ◽  
Essential Role ◽  
Nuclear Factor Kappa B ◽  
Ectodermal Dysplasia ◽  
Nuclear Factor ◽  
Nadph Oxidase Activity ◽  
Anhidrotic Ectodermal Dysplasia

scholarly journals Essential role of nuclear factor-κB for NADPH oxidase activity in normal and anhidrotic ectodermal dysplasia leukocytes

Blood ◽  
2008 ◽  
Vol 112 (4) ◽  
pp. 1453-1460 ◽  
Author(s):  
Marcos Luengo-Blanco ◽  
Carolina Prando ◽  
Jacinta Bustamante ◽  
Walmir Cutrim Aragão-Filho ◽  
Paulo Vitor Soeiro Pereira ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nadph Oxidase ◽  
Ectodermal Dysplasia ◽  
Nuclear Factor Κb ◽  
Nuclear Factor ◽  
U937 Cells ◽  
Cybb Gene ◽  
Anhidrotic Ectodermal Dysplasia ◽  
Ncf1 Gene

AbstractThis work investigated the functional role of nuclear factor–κB (NF-κB) in respiratory burst activity and in expression of the human phagocyte nicotinamide adenine dinucleotide phosphate (NADPH) oxidase genes CYBB, CYBA, NCF1, and NCF2. U937 cells with a stably transfected repressor of NF-κB (IκBα-S32A/S36A) demonstrated significantly lower superoxide release and lower CYBB and NCF1 gene expression compared with control U937 cells. We further tested Epstein-Barr virus (EBV)-transformed B cells from patients with anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID), an inherited disorder of NF-κB function. Superoxide release and CYBB gene expression by EDA-ID cells were significantly decreased compared with healthy cells and similar to cells from patients with X-linked chronic granulomatous disease (X910 CGD). NCF1 gene expression in EDA-ID S32I cells was decreased compared with healthy control cells and similar to that in autosomal recessive (A470) CGD cells. Gel shift assays demonstrated loss of recombinant human p50 binding to a NF-κB site 5′ to the CYBB gene in U937 cells treated with NF-κB inhibitors, repressor-transfected U937 cells, and EDA-ID patients' cells. Zymosan phagocytosis was not affected by transfection of U937 cells with the NF-κB repressor. These studies show that NF-κB is necessary for CYBB and NCF1 gene expression and activation of the phagocyte NADPH oxidase in this model system.

scholarly journals Nuclear factor-κB is not essential for NADPH oxidase activity in neutrophils from anhidrotic ectodermal dysplasia patients

Blood ◽  
2009 ◽  
Vol 113 (21) ◽  
pp. 5362-5363 ◽  
Author(s):  
Bram J. van Raam ◽  
Robin van Bruggen ◽  
Anton T. J. Tool ◽  
Machiel H. Jansen ◽  
Adilia Warris ◽  
...  
Keyword(s):  
Nadph Oxidase ◽  
Oxidase Activity ◽  
Ectodermal Dysplasia ◽  
Nuclear Factor Κb ◽  
Nuclear Factor ◽  
Nadph Oxidase Activity ◽  
Anhidrotic Ectodermal Dysplasia

scholarly journals IGF-I increases the expression of fibronectin by Nox4-dependent Akt phosphorylation in renal tubular epithelial cells

2012 ◽  
Vol 302 (1) ◽  
pp. C122-C130 ◽  
Author(s):  
David D. New ◽  
Karen Block ◽  
Basant Bhandhari ◽  
Yves Gorin ◽  
Hanna E. Abboud
Keyword(s):  
Epithelial Cells ◽  
Nadph Oxidase ◽  
Oxidase Activity ◽  
Dominant Negative ◽  
Tubular Epithelial Cells ◽  
Nadph Oxidase Activity ◽  
Akt Phosphorylation ◽  
Fibronectin Expression ◽  
Igf I

Extracellular matrix accumulation contributes to the progression of chronic kidney disease. Many growth factors including insulin-like growth factor-I (IGF-I) enhance matrix protein accumulation. Proximal tubular epithelial cells (PTCs) synthesize matrix proteins. NADPH oxidases are major sources of reactive oxygen species (ROS), important signaling molecules that mediate biological responses in a variety of cells and tissue. We investigated the mechanism by which IGF-I regulates fibronectin accumulation in PTCs and the role of a potential redox-dependent signaling pathway. IGF-I induces an increase in NADPH-dependent superoxide generation, enhances the release of hydrogen peroxide, and increases the expression of NADPH oxidase 4 (Nox4) in PTCs. IGF-I also stimulates phosphorylation of Akt, and inhibition of Akt or its upstream activator phosphatidylinositol 3-kinase attenuates IGF-I-induced fibronectin accumulation. Expression of dominant negative Akt also inhibits IGF-I-induced expression of fibronectin, indicating a role for this kinase in fibronectin accumulation. Expression of dominant negative adenovirus Nox4 inhibits IGF-I-induced NADPH oxidase activity, Akt phosphorylation, and fibronectin protein expression. Moreover, transfection of small interfering RNA targeting Nox4 decreases Nox4 protein expression and blocks IGF-I-induced Akt phosphorylation and the increase in fibronectin, placing Nox4 and ROS upstream of Akt signaling pathway. To confirm the role of Nox4, PTCs were infected with adenovirus construct expressing wild-type Nox4. Ad-Nox4, but not control Ad-green fluorescent protein, upregulated Nox4 expression and increased NADPH oxidase activity as well as fibronectin expression. Taken together, these results provide the first evidence for a role of Nox4 in IGF-I-induced Akt phosphorylation and fibronectin expression in tubular epithelial cells.

scholarly journals FcγR-stimulated activation of the NADPH oxidase: phosphoinositide-binding protein p40phox regulates NADPH oxidase activity after enzyme assembly on the phagosome

Blood ◽  
2008 ◽  
Vol 112 (9) ◽  
pp. 3867-3877 ◽  
Author(s):  
Wei Tian ◽  
Xing Jun Li ◽  
Natalie D. Stull ◽  
Wenyu Ming ◽  
Chang-Il Suh ◽  
...  
Keyword(s):  
Nadph Oxidase ◽  
Oxidase Activity ◽  
Superoxide Production ◽  
Resting Cells ◽  
Cellular Activation ◽  
Nadph Oxidase Activity ◽  
Phagocyte Nadph Oxidase ◽  
Before And After ◽  
Membrane Bound

AbstractThe phagocyte NADPH oxidase generates superoxide for microbial killing, and includes a membrane-bound flavocytochrome b558 and cytosolic p67phox, p47phox, and p40phox subunits that undergo membrane translocation upon cellular activation. The function of p40phox, which binds p67phox in resting cells, is incompletely understood. Recent studies showed that phagocytosis-induced superoxide production is stimulated by p40phox and its binding to phosphatidylinositol-3-phosphate (PI3P), a phosphoinositide enriched in membranes of internalized phagosomes. To better define the role of p40phox in FcγR-induced oxidase activation, we used immunofluorescence and real-time imaging of FcγR-induced phagocytosis. YFP-tagged p67phox and p40phox translocated to granulocyte phagosomes before phagosome internalization and accumulation of a probe for PI3P. p67phox and p47phox accumulation on nascent and internalized phagosomes did not require p40phox or PI3 kinase activity, although superoxide production before and after phagosome sealing was decreased by mutation of the p40phox PI3P-binding domain or wortmannin. Translocation of p40phox to nascent phagosomes required binding to p67phox but not PI3P, although the loss of PI3P binding reduced p40phox retention after phagosome internalization. We conclude that p40phox functions primarily to regulate FcγR-induced NADPH oxidase activity rather than assembly, and stimulates superoxide production via a PI3P signal that increases after phagosome internalization.

Apocynin administration prevents the changes induced by a fructose-rich diet on rat liver metabolism and the antioxidant system

2012 ◽  
Vol 123 (12) ◽  
pp. 681-692 ◽  
Author(s):  
María Cecilia Castro ◽  
Flavio Francini ◽  
Guillermo Schinella ◽  
Claudia Inés Caldiz ◽  
María Guillermina Zubiría ◽  
...  
Keyword(s):  
Gene Expression ◽  
Drinking Water ◽  
Nadph Oxidase ◽  
Protein Expression ◽  
Oxidase Activity ◽  
Fatty Acid Synthase ◽  
Control Diet ◽  
Liver Steatosis ◽  
Nadph Oxidase Activity ◽  
Gene And Protein Expression

In the present study, we investigated the role of NADPH oxidase in F (fructose)-rich-diet-induced hepatic OS (oxidative stress) and metabolic changes, and their prevention by apocynin co-administration. Wistar rats were fed for 21 days on (i) a control diet, (ii) a control diet plus 10% F in the drinking water, (iii) a control diet with apocynin in the drinking water (CA) and (iv) F plus apocynin in the drinking water (FA). Glycaemia, triglyceridaemia, NEFAs (non-esterified fatty acids) and insulinaemia were determined. In the liver, we measured (i) NADPH oxidase activity, and gene and protein expression; (ii) protein carbonyl groups, GSH and TBARSs (thiobarbituric acid-reactive substances); (iii) catalase, CuZn-SOD (superoxide dismutase) and Mn-SOD expression; (iv) liver glycogen and lipid content; (v) GK (glucokinase), G6Pase (glucose-6-phosphatase) and G6PDH (glucose-6-phosphate dehydrogenase) activities; (vi) FAS (fatty acid synthase), GPAT (glycerol-3-phosphate acyltransferase), G6Pase and G6PDH, IL-1β (interleukin-1β), PAI-1 (plasminogen-activator inhibitor-1) and TNFα (tumour necrosis factor α) gene expression; and (vii) IκBα (inhibitor of nuclear factor κB α) protein expression. F-fed animals had high serum TAG (triacylglycerol), NEFA and insulin levels, high liver NADPH oxidase activity/expression, increased OS markers, reduced antioxidant enzyme expression, and increased glycogen, TAG storage and GK, G6Pase and G6PDH activities. They also had high G6Pase, G6PDH, FAS, GPAT, TNFα and IL-1β gene expression and decreased IκBα expression. Co-administration of apocynin to F-fed rats prevented the development of most of these abnormalities. In conclusion, NADPH oxidase plays a key role in F-induced hepatic OS production and probably also in the mechanism of liver steatosis, suggesting its potential usefulness for the prevention/treatment of T2DM (Type 2 diabetes mellitus).

Novel Role of Protein Disulfide Isomerase in the Regulation of NADPH Oxidase Activity: Pathophysiological Implications in Vascular Diseases

2008 ◽  
Vol 10 (6) ◽  
pp. 1101-1114 ◽  
Author(s):  
Francisco R.M. Laurindo ◽  
Denise C. Fernandes ◽  
Angélica M. Amanso ◽  
Lucia R. Lopes ◽  
Célio X.C. Santos
Keyword(s):  
Nadph Oxidase ◽  
Oxidase Activity ◽  
Protein Disulfide Isomerase ◽  
Vascular Diseases ◽  
Disulfide Isomerase ◽  
Nadph Oxidase Activity ◽  
Protein Disulfide

scholarly journals Composition of partially purified NADPH oxidase from pig neutrophils

1984 ◽  
Vol 223 (3) ◽  
pp. 639-648 ◽  
Author(s):  
P Bellavite ◽  
O T G Jones ◽  
A R Cross ◽  
E Papini ◽  
F Rossi
Keyword(s):  
Enzyme Activity ◽  
Cytochrome C ◽  
Nadph Oxidase ◽  
Cytochrome B ◽  
Oxidase Activity ◽  
Gel Filtration ◽  
Purification Procedure ◽  
Nadph Oxidase Activity ◽  
Cytochrome C Reductase

The superoxide (O2.-)-forming enzyme NADPH oxidase from pig neutrophils was solubilized and partially purified by gel-filtration chromatography. The purification procedure allowed the separation of NADPH oxidase activity from NADH-dependent cytochrome c reductase and 2,6-dichlorophenol-indophenol reductase activities. O2.-forming activity was co-purified with cytochrome b-245 and was associated with phospholipids. However, active fractions endowed with cytochrome b were devoid of ubiquinone and contained only little FAD. The cytochrome b/FAD ratio was 1.13:1 in the crude solubilized extract and increased to 18.95:1 in the partially purified preparations. Most of FAD was associated with fractions containing NADH-dependent oxidoreductases. These results are consistent with the postulated role of cytochrome b in O2.-formation by neutrophil NADPH oxidase, but raise doubts about the participation of flavoproteins in this enzyme activity.

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