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Karl-heinz Krause - One of the best experts on this subject based on the ideXlab platform.
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role of nadph oxidase isoforms NOX1 nox2 and nox4 in myocardial ischemia reperfusion injury
Journal of Molecular and Cellular Cardiology, 2013Co-Authors: Vincent Braunersreuther, Fabrizio Montecucco, Mohammed Ashri, Graziano Pelli, Katia Galan, Miguel Frias, Fabienne Burger, Ana Luiza Gomez Quindere, Christophe Montessuit, Karl-heinz KrauseAbstract:Abstract Myocardial reperfusion injury is mediated by several processes including increase of reactive oxygen species (ROS). The aim of the study is to identify potential sources of ROS contributing to myocardial ischemia–reperfusion injury. For this purpose, we investigated myocardial ischemia/reperfusion pathology in mice deficient in various NADPH oxidase isoforms (NOX1, Nox2, Nox4, as well as NOX1/2 double knockout). Following 30 min of ischemia and 24 h of reperfusion, a significant decrease in the size of myocardial infarct was observed in NOX1-, Nox2- and NOX1/Nox2-, but not in Nox4-deficient mice. However, no protection was observed in a model of chronic ischemia, suggesting that NOX1 and NOX2-mediated oxidative damage occurs during reperfusion. Cardioprotective effect of NOX1 and Nox2 deficiencies was associated with decrease of neutrophil invasion, but, on the other hand an improved reperfusion injury was also observed in isolated perfused hearts (Langendorff model) suggesting that inflammatory cells were not the major source of oxidative damage. A decrease in global post-reperfusion oxidative stress was clearly detected in Nox2-, but not in NOX1-deficient hearts. Analysis of key signaling pathways during reperfusion suggests distinct cardioprotective patterns: increased phosphorylation was seen for Akt and Erk in NOX1-deficient mice and for Stat3 and Erk in Nox2-deficient mice. Consequently, NOX1 and NOX2 represent interesting drug targets for controlling reperfusion damage associated with revascularization in coronary disease.
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Role of NADPH oxidase isoforms NOX1, NOX2 and NOX4 in myocardial ischemia/reperfusion injury.
Journal of Molecular and Cellular Cardiology, 2013Co-Authors: Vincent Braunersreuther, Fabrizio Montecucco, Mohammed Ashri, Graziano Pelli, Katia Galan, Miguel Frias, Fabienne Burger, Ana Luiza Gomez Quindere, Christophe Montessuit, Karl-heinz KrauseAbstract:Abstract Myocardial reperfusion injury is mediated by several processes including increase of reactive oxygen species (ROS). The aim of the study is to identify potential sources of ROS contributing to myocardial ischemia–reperfusion injury. For this purpose, we investigated myocardial ischemia/reperfusion pathology in mice deficient in various NADPH oxidase isoforms (NOX1, Nox2, Nox4, as well as NOX1/2 double knockout). Following 30 min of ischemia and 24 h of reperfusion, a significant decrease in the size of myocardial infarct was observed in NOX1-, Nox2- and NOX1/Nox2-, but not in Nox4-deficient mice. However, no protection was observed in a model of chronic ischemia, suggesting that NOX1 and NOX2-mediated oxidative damage occurs during reperfusion. Cardioprotective effect of NOX1 and Nox2 deficiencies was associated with decrease of neutrophil invasion, but, on the other hand an improved reperfusion injury was also observed in isolated perfused hearts (Langendorff model) suggesting that inflammatory cells were not the major source of oxidative damage. A decrease in global post-reperfusion oxidative stress was clearly detected in Nox2-, but not in NOX1-deficient hearts. Analysis of key signaling pathways during reperfusion suggests distinct cardioprotective patterns: increased phosphorylation was seen for Akt and Erk in NOX1-deficient mice and for Stat3 and Erk in Nox2-deficient mice. Consequently, NOX1 and NOX2 represent interesting drug targets for controlling reperfusion damage associated with revascularization in coronary disease.
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NADPH Oxidase-1 Plays a Crucial Role in Hyperoxia-induced Acute Lung Injury in Mice
American Journal of Respiratory and Critical Care Medicine, 2009Co-Authors: Stephanie Carnesecchi, Christine Deffert, Alessandra Pagano, Isabelle Métrailler-ruchonnet, Michela G Schäppi, Yves Donati, Karl-heinz Krause, Sarah Garrido-urbani, Michael Matthay, Constance Barazzone ArgiroffoAbstract:Hyperoxia-induced acute lung injury has been used for many years as a model of oxidative stress mimicking clinical acute lung injury and the acute respiratory distress syndrome. Excess quantities of reactive oxygen species (ROS) are responsible for oxidative stress-induced lung injury. ROS are produced by mitochondrial chain transport, but also by NADPH oxidase (NOX) family members. Although NOX1 and NOX2 are expressed in the lungs, their precise function has not been determined until now.
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mechanisms of vascular smooth muscle nadph oxidase 1 NOX1 contribution to injury induced neointimal formation
Arteriosclerosis Thrombosis and Vascular Biology, 2009Co-Authors: Alejandra San Martin, David J Lambeth, Karl-heinz Krause, Botond Banfi, Anna Dikalova, Puja K Mehta, Abel Martin Garrido, Raju S Datla, Erin Lyons, Bernard LassegueAbstract:Objective— Vascular NADPH oxidases (Noxes) have been implicated in cardiovascular diseases; however, the importance of individual Nox homologues remains unclear. Here, the role of the vascular smooth muscle cell (VSMC) NOX1 in neointima formation was studied using genetically modified animal models. Methods and Results— Wire injury–induced neointima formation in the femoral artery, along with proliferation and apoptosis, was reduced in NOX1 y/− mice, but there was little difference in Tg SMCNOX1 mice compared with wild-type (WT) mice. Proliferation and migration were reduced in cultured NOX1 y/− VSMCs and increased in Tg SMCNOX1 cells. Tg SMCNOX1 cells exhibited increased fibronectin secretion, but neither collagen I production nor cell adhesion was affected by alteration of NOX1. Using antibody microarray and Western blotting analysis, increased cofilin phosphorylation and mDia1 expression and decreased PAK1 expression were detected in NOX1 y/− cells. Overexpression of S3A, a constitutively active cofilin mutant, partially recovered reduced migration of NOX1 y/− cells, suggesting that reduction in cofilin activity contributes to impaired migration of NOX1 y/− VSMCs. Conclusions— These results indicate that NOX1 plays a critical role in neointima formation by mediating VSMC migration, proliferation, and extracellular matrix production, and that cofilin is a major effector of NOX1-mediated migration. Inhibition of NOX1 may be an efficient strategy to suppress neointimal formation.
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NOX1 plays a crucial role in hyperoxia-induced acute lung injury in mice
2009Co-Authors: Stephanie Carnesecchi, Christine Deffert, Alessandra Pagano, Sarah Garrido, Isabelle Métrailler-ruchonnet, Michela G Schäppi, Yves Donati, Karl-heinz Krause, Constance Barazzone Argiroffo, Carnesecchi StephanieAbstract:Figure E1. Expression of NOX isoforms in NOX1and NOX2-deficient mice. NOX1 (A), NOX2 (B) and NOX4 (C) lung mRNA expression was measured in NOX1 -/, NOX2 -/and WT mice by real time RT-PCR in air condition. (n=3 mice in each group, P=NS, NOX2 and NOX4 mRNA expression in NOX1 -/versus WT mice, and NOX1 and NOX4 mRNA expression in NOX2 -/versus WT mice. To notice that there is no compensatory expression of NOX isoforms in all deficient mice.
David J Lambeth - One of the best experts on this subject based on the ideXlab platform.
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mechanisms of vascular smooth muscle nadph oxidase 1 NOX1 contribution to injury induced neointimal formation
Arteriosclerosis Thrombosis and Vascular Biology, 2009Co-Authors: Alejandra San Martin, David J Lambeth, Karl-heinz Krause, Botond Banfi, Anna Dikalova, Puja K Mehta, Abel Martin Garrido, Raju S Datla, Erin Lyons, Bernard LassegueAbstract:Objective— Vascular NADPH oxidases (Noxes) have been implicated in cardiovascular diseases; however, the importance of individual Nox homologues remains unclear. Here, the role of the vascular smooth muscle cell (VSMC) NOX1 in neointima formation was studied using genetically modified animal models. Methods and Results— Wire injury–induced neointima formation in the femoral artery, along with proliferation and apoptosis, was reduced in NOX1 y/− mice, but there was little difference in Tg SMCNOX1 mice compared with wild-type (WT) mice. Proliferation and migration were reduced in cultured NOX1 y/− VSMCs and increased in Tg SMCNOX1 cells. Tg SMCNOX1 cells exhibited increased fibronectin secretion, but neither collagen I production nor cell adhesion was affected by alteration of NOX1. Using antibody microarray and Western blotting analysis, increased cofilin phosphorylation and mDia1 expression and decreased PAK1 expression were detected in NOX1 y/− cells. Overexpression of S3A, a constitutively active cofilin mutant, partially recovered reduced migration of NOX1 y/− cells, suggesting that reduction in cofilin activity contributes to impaired migration of NOX1 y/− VSMCs. Conclusions— These results indicate that NOX1 plays a critical role in neointima formation by mediating VSMC migration, proliferation, and extracellular matrix production, and that cofilin is a major effector of NOX1-mediated migration. Inhibition of NOX1 may be an efficient strategy to suppress neointimal formation.
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redox dependent expression of cyclin d1 and cell proliferation by NOX1 in mouse lung epithelial cells
Antioxidants & Redox Signaling, 2006Co-Authors: Priya Ranjan, David J Lambeth, Vikas Anathy, Peter M Burch, Kelly Weirather, Nicholas H HeintzAbstract:NADPH oxidases produce reactive oxygen species (ROS) that serve as co-stimulatory signals for cell proliferation. In mouse lung epithelial cells that express NOX1, Nox2, Nox4, p22phox, p47phox, p67phox, and Noxo1, overexpression of NOX1 delayed cell cycle withdrawal by maintaining AP-1-dependent expression of cyclin D1 in low serum conditions. In cycling cells, the effects of NOX1 were dose dependent: levels of NOX1 that induced 3- to 10-fold increases in ROS promoted phosphorylation of ERK1/2 and expression of cyclin D1, whereas expression of NOX1 with Noxo1 and Noxa1 (or expression of Nox4 alone) that induced substantial increases in intracellular ROS inhibited cyclin D1 and proliferation. Catalase reversed the effects of NOX1 on cyclin D1 and cell proliferation. Diphenylene iodonium, an inhibitor of NADPH oxidase activity, did not affect dosedependent responses of ERK1/2 or Akt to serum, but markedly inhibited the sequential expression of c-Fos and Fra-1 required for induction of cyclin D1 during cell ...
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NOX1 dependent reactive oxygen generation is regulated by rac1
Journal of Biological Chemistry, 2006Co-Authors: Guangjie Cheng, Becky A Diebold, Yasmin Hughes, David J LambethAbstract:Abstract Rac1 has been implicated in the generation of reactive oxygen species (ROS) in several cell types, but the enzymatic origin of the ROS has not been proven. The present studies demonstrate that NOX1, a homolog of the phagocyte NADPH-oxidase component gp91phox, is activated by Rac1. When NOX1 is co-expressed along with its regulatory subunits NOXO1 and NOXA1, significant ROS generation is seen. Herein, co-expression of constitutively active Rac1(G12V), but not wild-type Rac1, resulted in marked further stimulation of activity. Decreased Rac1 expression using small interfering RNA reduced NOX1-dependent ROS. CDC42(G12V) failed to increase activity, and small interfering RNA directed against CDC42 failed to decrease activity, pointing to specificity for Rac. TPR domain mutants of NOXA1 that interfere with Rac1 binding were ineffective in supporting NOX1-dependent ROS generation. Immunoprecipitation experiments demonstrated a complex containing Rac1(G12V), NOXO1, NOXA1, and NOX1. CDC42(G12V) could not substitute for Rac1(G12V) in such a complex. NOX1 formed a complex with Rac1(G12V) that was independent of NOXA1 and NOXO1, consistent with direct binding of Rac1(G12V) to NOX1. Rac1(G12V) interaction with NOXA1 was enhanced by NOX1 and NOXO1, suggesting cooperative binding. A model is presented comparing activation by regulatory subunits of NOX1 versus gp91phox (Nox2) in which Rac1 activation provides a major trigger that acutely activates NOX1-dependent ROS generation.
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point mutations in the proline rich region of p22phox are dominant inhibitors of NOX1 and nox2 dependent reactive oxygen generation
Journal of Biological Chemistry, 2005Co-Authors: Tsukasa Kawahara, Darren R Ritsick, Guangjie Cheng, David J LambethAbstract:Abstract The integral membrane protein p22phox is an indispensable component of the superoxide-generating phagocyte NADPH oxidase, whose catalytic core is the membrane-associated gp91phox (also known as Nox2). p22phox associates with gp91phox and, through its proline-rich C terminus, provides a binding site for the tandem Src homology 3 domains of the activating subunit p47phox. Whereas p22phox is expressed ubiquitously, its participation in regulating the activity of other Nox enzymes is less clear. This study investigates the requirement of p22phox for Nox enzyme activity and explores the role of its proline-rich region (PRR) for regulating activity. Coexpression of specific Nox catalytic subunits (NOX1, Nox2, Nox3, Nox4, or Nox5) along with their corresponding regulatory subunits (NOXO1/NOXA1 for NOX1; p47phox/p67phox/Rac for Nox2; NOXO1 for Nox3; no subunits for Nox4 or Nox5) resulted in marked production of reactive oxygen. Small interfering RNAs decreased endogenous p22phox expression and inhibited reactive oxygen generation from NOX1, Nox2, Nox3, and Nox4 but not Nox5. Truncated forms of p22phox that disrupted the PRR-inhibited reactive oxygen generation from NOX1, Nox2, and Nox3 but not from Nox4 and Nox5. Similarly, p22phox (P156Q), a mutation that disrupts Src homology 3 binding by the PRR, potently inhibited reactive oxygen production from NOX1 and Nox2 but not from Nox4 and Nox5. Expression of p22phox (P156Q) inhibited NOXO1-stimulated Nox3 activity, but co-expression of NOXA1 overcame the inhibitory effect. The P157Q and P160Q mutations of p22phox showed selective inhibition of Nox2/p47phox/p67phox, and selectivity was specific for the organizing subunit (p47phox or NOXO1) rather than the Nox catalytic subunit. These studies stress the importance of p22phox for the function of NOX1, Nox2, Nox3, and Nox4, and emphasize the key role of the PRR for regulating Nox proteins whose activity is dependent upon p47phox or NOXO1.
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increased NOX1 and hydrogen peroxide in prostate cancer
The Prostate, 2005Co-Authors: David J Lambeth, Fray F Marshall, Mahul B Amin, Leland W K Chung, John A Petros, Rebecca S ArnoldAbstract:BACKGROUND Reactive oxygen species (ROS) are emerging as candidate mediators of growth and angiogenesis in cancer. Increased ROS often correlates with cell growth, e.g., Ras-transformed cells and cells treated with growth factors. While non-transformed cells respond to growth factors/cytokines with the regulated production of ROS, tumor cells in culture frequently overproduce H2O2. We propose that NADPH oxidases (Nox) account for increased levels of ROS in some cancers. Previously, transfection of NOX1 into a prostate cancer cell line dramatically enhanced tumor growth (Arbiser et al.: PNAS 99:715–720, 2001). METHODS Using immunohistochemistry, immunofluorescence, dihydroethidium staining, and Flow cytometry, we investigated the correlation between NOX1 and ROS in prostate cancer. RESULTS Here, we demonstrate that human prostate tumors show increased H2O2 levels. Furthermore, 80% of human prostate tumor samples show markedly increased NOX1 protein levels and increased mRNA levels. In addition, a series of cell lines developed from LNCaP prostate cancer cells that demonstrate increasing tumor and metastatic potential, show increased NOX1 and a parallel increase in H2O2 levels. CONCLUSIONS The results illustrate that human prostate cancer frequently show both increased H2O2 and NOX1, and that in an animal model system increased NOX1/H2O2 correlates with increased tumorigenicity. © 2004 Wiley-Liss, Inc.
Chihiro Yabenishimura - One of the best experts on this subject based on the ideXlab platform.
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depressive like behaviors are regulated by NOX1 nadph oxidase by redox modification of nmda receptor 1
The Journal of Neuroscience, 2017Co-Authors: Noriaki Arakawa, Misaki Matsumoto, Masato Katsuyama, Kazumi Iwata, Shiho Kitaoka, Ai Kawaji, Kenichi Matsuda, Satoshi Teramukai, Tomoyuki Furuyashiki, Chihiro YabenishimuraAbstract:Involvement of reactive oxygen species (ROS) has been suggested in the development of psychiatric disorders. NOX1 is a nonphagocytic form of NADPH oxidase whose expression in the nervous system is negligible compared with other NOX isoforms. However, NOX1-derived ROS increase inflammatory pain and tolerance to opioid analgesia. To clarify the role of NOX1 in the brain, we examined depressive-like behaviors in mice deficient in NOX1 ( NOX1 −/Y). Depressive-like behaviors induced by chronic social defeat stress or administration of corticosterone (CORT) were significantly ameliorated in NOX1 −/Y. Generation of ROS was significantly elevated in the prefrontal cortex (PFC) of mice administrated with CORT, while NOX1 mRNA was upregulated only in the ventral tegmental area (VTA) among brain areas responsible for emotional behaviors. Delivery of miRNA against NOX1 to VTA restored CORT-induced depressive-like behaviors in wild-type (WT) littermates. Administration of CORT to WT, but not to NOX1 −/Y, significantly reduced transcript levels of brain-derived neurotrophic factor ( bdnf ), with a concomitant increase in DNA methylation of the promoter regions in bdnf . Delivery of miRNA against NOX1 to VTA restored the level of BDNF mRNA in WT PFC. Redox proteome analyses demonstrated that NMDA receptor 1 (NR1) was among the molecules redox regulated by NOX1. In cultured cortical neurons, hydrogen peroxide significantly suppressed NMDA-induced upregulation of BDNF transcripts in NR1-expressing cells but not in cells harboring mutant NR1 (C744A). Together, these findings suggest a key role of NOX1 in depressive-like behaviors through NR1-mediated epigenetic modification of bdnf in the mesoprefrontal projection. SIGNIFICANCE STATEMENT NADPH oxidase is a source of reactive oxygen species (ROS) that have been implicated in the pathogenesis of various neurological disorders. We presently showed the involvement of a nonphagocytic type of NADPH oxidase, NOX1, in major depressive disorders, including behavioral, biochemical, and anatomical changes in mice. The oxidation of NR1 by NOX1-derived ROS was demonstrated in prefrontal cortex (PFC), which may be causally linked to the downregulation of BDNF, promoting depressive-like behaviors. Given that NOX1 is upregulated only in VTA but not in PFC, mesocortical projections appear to play a crucial role in NOX1-dependent depressive-like behaviors. Our study is the first to present the potential molecular mechanism underlying the development of major depression through the NOX1-induced oxidation of NR1 and epigenetic modification of bdnf .
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the ros generating oxidase NOX1 is required for epithelial restitution following colitis
Experimental Animals, 2016Co-Authors: Masayoshi Kato, Chihiro Yabenishimura, Misaki Matsumoto, Masaya Marumo, Jun Nakayama, Tohru KamataAbstract:Accumulating evidence suggests that reactive oxygen species (ROS) generated by endogenous metabolic enzymes are involved in a variety of intracellular mechanisms. In particular, superoxide-generating NADPH oxidase (Nox) 1 is highly expressed in the colon and has been implicated in physiological and pathophysiological states of colon tissues. However, its role in tissue repair following colitis has not been fully elucidated. Our study using experimental colitis in mice showed that repair of the mucosal layer did not occur in NOX1-deficient mice following dextran sulfate sodium-induced colitis. This was accompanied by inhibition of proliferation, cell survival, migration, and terminal differentiation (generation of goblet cells) of crypt progenitor cells, as determined by histochemical analyses. Furthermore, NOX1 expression as well as ROS production in the colon crypt was increased during the repair process, and NOX1 deficiency suppressed these events. The results suggest that NOX1 promotes colon mucosal wound repair by sustaining the bioactivity of crypt progenitor cells and plays a crucial role in the epithelial restitution in the case of damage associated with colitis.
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ros generating oxidases NOX1 and nox4 contribute to oncogenic ras induced premature senescence
Genes to Cells, 2013Co-Authors: Ryo Kodama, Masayoshi Kato, Shuichi Furuta, Shouta Ueno, Yugen Zhang, Kuniharu Matsuno, Chihiro Yabenishimura, Eiji Tanaka, Tohru KamataAbstract:Activated oncogenes induce premature cellular senescence, a permanent state of proliferative arrest in primary rodent and human fibroblasts. Recent studies suggest that generation of reactive oxygen species (ROS) is involved in oncogenic Ras-induced premature senescence. However, the signaling mechanism controlling this oxidant-mediated irreversible growth arrest is not fully understood. Here, we show that through the Ras/MEK pathway, Ras oncogene up-regulated the expression of superoxide-generating oxidases, NOX1 in rat REF52 cells and Nox4 in primary human lung TIG-3 cells, leading to an increase in intracellular level of ROS. Ablation of NOX1 and Nox4 by small interfering RNAs (siRNAs) blocked the RasV12 senescent phenotype including β-galactosidase activity, growth arrest and accumulation of tumor suppressors such as p53 and p16Ink4a. This suggests that Nox-generated ROS transduce senescence signals by activating the p53 and p16Ink4a pathway. Furthermore, NOX1 and Nox4 siRNAs inhibited both Ras-induced DNA damage response and p38MAPK activation, whereas overexpression of NOX1 and Nox4 alone was able to induce senescence. The involvement of NOX1 in Ras-induced senescence was also confirmed with embryonic fibroblasts derived from NOX1 knockout mice. Together, these findings suggest that NOX1- and Nox4-generated ROS play an important role in Ras-induced premature senescence, which may involve DNA damage response and p38MAPK signaling pathways.
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NOX1 nicotinamide adenine dinucleotide phosphate reduced form nadph oxidase promotes proliferation of stellate cells and aggravates liver fibrosis induced by bile duct ligation
Hepatology, 2011Co-Authors: Kuniharu Matsuno, Misaki Matsumoto, Jia Zhang, Masato Katsuyama, Kazumi Iwata, Natalie J Torok, Chihiro YabenishimuraAbstract:Among multiple isoforms of nicotinamide adenine dinucleotide phosphate, reduced form (NADPH) oxidase expressed in the liver, the phagocytic NOX2 isoform in hepatic stellate cells (HSCs) has been demonstrated to play a key role in liver fibrogenesis. The aim of this study was to clarify the role of NOX1, a nonphagocytic form of NADPH oxidase, in the development of fibrosis using NOX1-deficient mice (NOX1KO). Liver injury and fibrosis were induced by bile duct ligation (BDL) and carbon tetrachloride in NOX1KO and wildtype littermate mice (WT). Primary HSCs were isolated to characterize the NOX1-induced signaling cascade involved in liver fibrogenesis. Following BDL, a time-dependent increase in NOX1 messenger RNA (mRNA) was demonstrated in WT liver. Compared with those in WT, levels of collagen-1α mRNA and hydroxyproline were significantly suppressed in NOX1KO with a reduced number of activated HSCs and less severe fibrotic lesions. The expression levels of α-smooth muscle actin, a marker of HSCs activation, were similar in cultured HSCs isolated from both genotypes. However, cell proliferation was significantly attenuated in HSCs isolated from NOX1KO. In these cells, the expression of p27kip1, a cell cycle suppressor, was significantly up-regulated. Concomitantly, a significant reduction in phosphorylated forms of Akt and forkhead box O (FOXO) 4, a downstream effector of Akt that regulates the transcription of p27kip1 gene, was demonstrated in NOX1KO. Finally, the level of the oxidized inactivated form of phosphatase and tensin homolog (PTEN), a negative regulator of PI3K/Akt pathway, was significantly attenuated in HSCs of NOX1KO. Conclusion: These findings indicate that reactive oxygen species derived from NOX1/NADPH oxidase oxidize and inactivate PTEN to positively regulate the Akt/FOXO4/p27kip1 signaling pathway. NOX1 may thus promote proliferation of HSCs and accelerate the development of fibrosis following BDL-induced liver injury. (HEPATOLOGY 2011;)
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NOX1 is involved in angiotensin ii mediated hypertension a study in NOX1 deficient mice
Circulation, 2005Co-Authors: Kuniharu Matsuno, Masato Katsuyama, Kazumi Iwata, Hiroyuki Yamada, Masato Matsuki, Shinji Takai, Kiyofumi Yamanishi, Mizuo Miyazaki, Hiroaki Matsubara, Chihiro YabenishimuraAbstract:Background— Increased production of reactive oxygen species (ROSs) by angiotensin II (Ang II) is involved in the initiation and progression of cardiovascular diseases. NADPH oxidase is a major source of superoxide generated in vascular tissues. Although NOX1 has been identified in vascular smooth muscle cells as a new homolog of gp91phox (Nox2), a catalytic subunit of NADPH oxidase, the pathophysiological function of NOX1-derived ROSs has not been fully elucidated. To clarify the role of NOX1 in Ang II–mediated hypertension, we generated NOX1-deficient (−/Y) mice. Methods and Results— No difference in the baseline blood pressure was observed between NOX1+/Y and NOX1−/Y. Infusion of Ang II induced a significant increase in mean blood pressure, accompanied by augmented expression of NOX1 mRNA and superoxide production in the aorta of NOX1+/Y, whereas the elevation in blood pressure and production of superoxide were significantly blunted in NOX1−/Y. Conversely, the infusion of pressor as well as subpressor d...
Kuniharu Matsuno - One of the best experts on this subject based on the ideXlab platform.
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ros generating oxidases NOX1 and nox4 contribute to oncogenic ras induced premature senescence
Genes to Cells, 2013Co-Authors: Ryo Kodama, Masayoshi Kato, Shuichi Furuta, Shouta Ueno, Yugen Zhang, Kuniharu Matsuno, Chihiro Yabenishimura, Eiji Tanaka, Tohru KamataAbstract:Activated oncogenes induce premature cellular senescence, a permanent state of proliferative arrest in primary rodent and human fibroblasts. Recent studies suggest that generation of reactive oxygen species (ROS) is involved in oncogenic Ras-induced premature senescence. However, the signaling mechanism controlling this oxidant-mediated irreversible growth arrest is not fully understood. Here, we show that through the Ras/MEK pathway, Ras oncogene up-regulated the expression of superoxide-generating oxidases, NOX1 in rat REF52 cells and Nox4 in primary human lung TIG-3 cells, leading to an increase in intracellular level of ROS. Ablation of NOX1 and Nox4 by small interfering RNAs (siRNAs) blocked the RasV12 senescent phenotype including β-galactosidase activity, growth arrest and accumulation of tumor suppressors such as p53 and p16Ink4a. This suggests that Nox-generated ROS transduce senescence signals by activating the p53 and p16Ink4a pathway. Furthermore, NOX1 and Nox4 siRNAs inhibited both Ras-induced DNA damage response and p38MAPK activation, whereas overexpression of NOX1 and Nox4 alone was able to induce senescence. The involvement of NOX1 in Ras-induced senescence was also confirmed with embryonic fibroblasts derived from NOX1 knockout mice. Together, these findings suggest that NOX1- and Nox4-generated ROS play an important role in Ras-induced premature senescence, which may involve DNA damage response and p38MAPK signaling pathways.
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ROS‐generating oxidases NOX1 and Nox4 contribute to oncogenic Ras‐induced premature senescence
Genes to Cells, 2012Co-Authors: Ryo Kodama, Masayoshi Kato, Shuichi Furuta, Shouta Ueno, Yugen Zhang, Kuniharu Matsuno, Eiji Tanaka, Chihiro Yabe-nishimura, Tohru KamataAbstract:Activated oncogenes induce premature cellular senescence, a permanent state of proliferative arrest in primary rodent and human fibroblasts. Recent studies suggest that generation of reactive oxygen species (ROS) is involved in oncogenic Ras-induced premature senescence. However, the signaling mechanism controlling this oxidant-mediated irreversible growth arrest is not fully understood. Here, we show that through the Ras/MEK pathway, Ras oncogene up-regulated the expression of superoxide-generating oxidases, NOX1 in rat REF52 cells and Nox4 in primary human lung TIG-3 cells, leading to an increase in intracellular level of ROS. Ablation of NOX1 and Nox4 by small interfering RNAs (siRNAs) blocked the RasV12 senescent phenotype including β-galactosidase activity, growth arrest and accumulation of tumor suppressors such as p53 and p16Ink4a. This suggests that Nox-generated ROS transduce senescence signals by activating the p53 and p16Ink4a pathway. Furthermore, NOX1 and Nox4 siRNAs inhibited both Ras-induced DNA damage response and p38MAPK activation, whereas overexpression of NOX1 and Nox4 alone was able to induce senescence. The involvement of NOX1 in Ras-induced senescence was also confirmed with embryonic fibroblasts derived from NOX1 knockout mice. Together, these findings suggest that NOX1- and Nox4-generated ROS play an important role in Ras-induced premature senescence, which may involve DNA damage response and p38MAPK signaling pathways.
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NOX1 nadph oxidase is involved in endotoxin induced cardiomyocyte apoptosis
Free Radical Biology and Medicine, 2012Co-Authors: Kuniharu Matsuno, Misaki Matsumoto, Jia Zhang, Masato Katsuyama, Kazumi Iwata, Ayumi Murata, Hideo Nakamura, Kanako Ikami, Satoaki Matoba, Shinji TakaiAbstract:Abstract The functional significance of NOX1/NADPH oxidase in the heart has not been explored due to its low expression relative to other NOX homologs identified so far. We aimed to clarify the role of NOX1/NADPH oxidase in the septic heart by utilizing mice deficient in the NOX1 gene (NOX1 −/Y ). Sepsis was induced by intraperitoneal administration of lipopolysaccharides (LPS: 25 mg/kg) or cecal ligation and puncture (CLP) surgery. A marked elevation of NOX1 mRNA was demonstrated in cardiac tissue, which was accompanied by increased production of reactive oxygen species (ROS). In NOX1 −/Y treated with LPS, cardiac dysfunction and survival were significantly improved compared with wild-type mice (NOX1 +/Y ) treated with LPS. Concomitantly, LPS-induced cardiomyocyte apoptosis and activation of caspase-3 were alleviated in NOX1 −/Y . The level of phosphorylated Akt in cardiac tissue was significantly lowered in NOX1 +/Y but not in NOX1 −/Y treated with LPS or that underwent CLP surgery. Increased oxidation of cysteine residues of Akt and enhanced interaction of Akt with protein phosphatase 2A (PP2A), a major phosphatase implicated in the dephosphorylation of Akt, were demonstrated in LPS-treated NOX1 +/Y . These responses to LPS were significantly attenuated in NOX1 −/Y . Taken together, ROS derived from NOX1/NADPH oxidase play a pivotal role in endotoxin-induced cardiomyocyte apoptosis by increasing oxidation of Akt and subsequent dephosphorylation by PP2A. Marked up-regulation of NOX1 may affect the risk of mortality under systemic inflammatory conditions.
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NOX1/NADPH oxidase is involved in endotoxin-induced cardiomyocyte apoptosis
Free Radical Biology and Medicine, 2012Co-Authors: Kuniharu Matsuno, Misaki Matsumoto, Masato Katsuyama, Kazumi Iwata, Ayumi Murata, Hideo Nakamura, Kanako Ikami, Jia ZhangAbstract:Abstract The functional significance of NOX1/NADPH oxidase in the heart has not been explored due to its low expression relative to other NOX homologs identified so far. We aimed to clarify the role of NOX1/NADPH oxidase in the septic heart by utilizing mice deficient in the NOX1 gene (NOX1 −/Y ). Sepsis was induced by intraperitoneal administration of lipopolysaccharides (LPS: 25 mg/kg) or cecal ligation and puncture (CLP) surgery. A marked elevation of NOX1 mRNA was demonstrated in cardiac tissue, which was accompanied by increased production of reactive oxygen species (ROS). In NOX1 −/Y treated with LPS, cardiac dysfunction and survival were significantly improved compared with wild-type mice (NOX1 +/Y ) treated with LPS. Concomitantly, LPS-induced cardiomyocyte apoptosis and activation of caspase-3 were alleviated in NOX1 −/Y . The level of phosphorylated Akt in cardiac tissue was significantly lowered in NOX1 +/Y but not in NOX1 −/Y treated with LPS or that underwent CLP surgery. Increased oxidation of cysteine residues of Akt and enhanced interaction of Akt with protein phosphatase 2A (PP2A), a major phosphatase implicated in the dephosphorylation of Akt, were demonstrated in LPS-treated NOX1 +/Y . These responses to LPS were significantly attenuated in NOX1 −/Y . Taken together, ROS derived from NOX1/NADPH oxidase play a pivotal role in endotoxin-induced cardiomyocyte apoptosis by increasing oxidation of Akt and subsequent dephosphorylation by PP2A. Marked up-regulation of NOX1 may affect the risk of mortality under systemic inflammatory conditions.
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NOX1 nicotinamide adenine dinucleotide phosphate reduced form nadph oxidase promotes proliferation of stellate cells and aggravates liver fibrosis induced by bile duct ligation
Hepatology, 2011Co-Authors: Kuniharu Matsuno, Misaki Matsumoto, Jia Zhang, Masato Katsuyama, Kazumi Iwata, Natalie J Torok, Chihiro YabenishimuraAbstract:Among multiple isoforms of nicotinamide adenine dinucleotide phosphate, reduced form (NADPH) oxidase expressed in the liver, the phagocytic NOX2 isoform in hepatic stellate cells (HSCs) has been demonstrated to play a key role in liver fibrogenesis. The aim of this study was to clarify the role of NOX1, a nonphagocytic form of NADPH oxidase, in the development of fibrosis using NOX1-deficient mice (NOX1KO). Liver injury and fibrosis were induced by bile duct ligation (BDL) and carbon tetrachloride in NOX1KO and wildtype littermate mice (WT). Primary HSCs were isolated to characterize the NOX1-induced signaling cascade involved in liver fibrogenesis. Following BDL, a time-dependent increase in NOX1 messenger RNA (mRNA) was demonstrated in WT liver. Compared with those in WT, levels of collagen-1α mRNA and hydroxyproline were significantly suppressed in NOX1KO with a reduced number of activated HSCs and less severe fibrotic lesions. The expression levels of α-smooth muscle actin, a marker of HSCs activation, were similar in cultured HSCs isolated from both genotypes. However, cell proliferation was significantly attenuated in HSCs isolated from NOX1KO. In these cells, the expression of p27kip1, a cell cycle suppressor, was significantly up-regulated. Concomitantly, a significant reduction in phosphorylated forms of Akt and forkhead box O (FOXO) 4, a downstream effector of Akt that regulates the transcription of p27kip1 gene, was demonstrated in NOX1KO. Finally, the level of the oxidized inactivated form of phosphatase and tensin homolog (PTEN), a negative regulator of PI3K/Akt pathway, was significantly attenuated in HSCs of NOX1KO. Conclusion: These findings indicate that reactive oxygen species derived from NOX1/NADPH oxidase oxidize and inactivate PTEN to positively regulate the Akt/FOXO4/p27kip1 signaling pathway. NOX1 may thus promote proliferation of HSCs and accelerate the development of fibrosis following BDL-induced liver injury. (HEPATOLOGY 2011;)
Kazumi Iwata - One of the best experts on this subject based on the ideXlab platform.
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depressive like behaviors are regulated by NOX1 nadph oxidase by redox modification of nmda receptor 1
The Journal of Neuroscience, 2017Co-Authors: Noriaki Arakawa, Misaki Matsumoto, Masato Katsuyama, Kazumi Iwata, Shiho Kitaoka, Ai Kawaji, Kenichi Matsuda, Satoshi Teramukai, Tomoyuki Furuyashiki, Chihiro YabenishimuraAbstract:Involvement of reactive oxygen species (ROS) has been suggested in the development of psychiatric disorders. NOX1 is a nonphagocytic form of NADPH oxidase whose expression in the nervous system is negligible compared with other NOX isoforms. However, NOX1-derived ROS increase inflammatory pain and tolerance to opioid analgesia. To clarify the role of NOX1 in the brain, we examined depressive-like behaviors in mice deficient in NOX1 ( NOX1 −/Y). Depressive-like behaviors induced by chronic social defeat stress or administration of corticosterone (CORT) were significantly ameliorated in NOX1 −/Y. Generation of ROS was significantly elevated in the prefrontal cortex (PFC) of mice administrated with CORT, while NOX1 mRNA was upregulated only in the ventral tegmental area (VTA) among brain areas responsible for emotional behaviors. Delivery of miRNA against NOX1 to VTA restored CORT-induced depressive-like behaviors in wild-type (WT) littermates. Administration of CORT to WT, but not to NOX1 −/Y, significantly reduced transcript levels of brain-derived neurotrophic factor ( bdnf ), with a concomitant increase in DNA methylation of the promoter regions in bdnf . Delivery of miRNA against NOX1 to VTA restored the level of BDNF mRNA in WT PFC. Redox proteome analyses demonstrated that NMDA receptor 1 (NR1) was among the molecules redox regulated by NOX1. In cultured cortical neurons, hydrogen peroxide significantly suppressed NMDA-induced upregulation of BDNF transcripts in NR1-expressing cells but not in cells harboring mutant NR1 (C744A). Together, these findings suggest a key role of NOX1 in depressive-like behaviors through NR1-mediated epigenetic modification of bdnf in the mesoprefrontal projection. SIGNIFICANCE STATEMENT NADPH oxidase is a source of reactive oxygen species (ROS) that have been implicated in the pathogenesis of various neurological disorders. We presently showed the involvement of a nonphagocytic type of NADPH oxidase, NOX1, in major depressive disorders, including behavioral, biochemical, and anatomical changes in mice. The oxidation of NR1 by NOX1-derived ROS was demonstrated in prefrontal cortex (PFC), which may be causally linked to the downregulation of BDNF, promoting depressive-like behaviors. Given that NOX1 is upregulated only in VTA but not in PFC, mesocortical projections appear to play a crucial role in NOX1-dependent depressive-like behaviors. Our study is the first to present the potential molecular mechanism underlying the development of major depression through the NOX1-induced oxidation of NR1 and epigenetic modification of bdnf .
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Depressive-Like Behaviors Are Regulated by NOX1/NADPH Oxidase by Redox Modification of NMDA Receptor 1
The Journal of Neuroscience, 2017Co-Authors: Noriaki Arakawa, Misaki Matsumoto, Kazumi Iwata, Shiho Kitaoka, Ai Kawaji, Kenichi Matsuda, Masato KatsuyamaAbstract:Involvement of reactive oxygen species (ROS) has been suggested in the development of psychiatric disorders. NOX1 is a nonphagocytic form of NADPH oxidase whose expression in the nervous system is negligible compared with other NOX isoforms. However, NOX1-derived ROS increase inflammatory pain and tolerance to opioid analgesia. To clarify the role of NOX1 in the brain, we examined depressive-like behaviors in mice deficient in NOX1 ( NOX1 −/Y). Depressive-like behaviors induced by chronic social defeat stress or administration of corticosterone (CORT) were significantly ameliorated in NOX1 −/Y. Generation of ROS was significantly elevated in the prefrontal cortex (PFC) of mice administrated with CORT, while NOX1 mRNA was upregulated only in the ventral tegmental area (VTA) among brain areas responsible for emotional behaviors. Delivery of miRNA against NOX1 to VTA restored CORT-induced depressive-like behaviors in wild-type (WT) littermates. Administration of CORT to WT, but not to NOX1 −/Y, significantly reduced transcript levels of brain-derived neurotrophic factor ( bdnf ), with a concomitant increase in DNA methylation of the promoter regions in bdnf . Delivery of miRNA against NOX1 to VTA restored the level of BDNF mRNA in WT PFC. Redox proteome analyses demonstrated that NMDA receptor 1 (NR1) was among the molecules redox regulated by NOX1. In cultured cortical neurons, hydrogen peroxide significantly suppressed NMDA-induced upregulation of BDNF transcripts in NR1-expressing cells but not in cells harboring mutant NR1 (C744A). Together, these findings suggest a key role of NOX1 in depressive-like behaviors through NR1-mediated epigenetic modification of bdnf in the mesoprefrontal projection. SIGNIFICANCE STATEMENT NADPH oxidase is a source of reactive oxygen species (ROS) that have been implicated in the pathogenesis of various neurological disorders. We presently showed the involvement of a nonphagocytic type of NADPH oxidase, NOX1, in major depressive disorders, including behavioral, biochemical, and anatomical changes in mice. The oxidation of NR1 by NOX1-derived ROS was demonstrated in prefrontal cortex (PFC), which may be causally linked to the downregulation of BDNF, promoting depressive-like behaviors. Given that NOX1 is upregulated only in VTA but not in PFC, mesocortical projections appear to play a crucial role in NOX1-dependent depressive-like behaviors. Our study is the first to present the potential molecular mechanism underlying the development of major depression through the NOX1-induced oxidation of NR1 and epigenetic modification of bdnf .
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Characterization of N-glycosylation sites on the extracellular domain of NOX1/NADPH oxidase.
Free Radical Biology and Medicine, 2013Co-Authors: Misaki Matsumoto, William M Nauseef, Jia Zhang, Masato Katsuyama, Kazumi Iwata, Chihiro Yabe-nishimuraAbstract:Abstract Extensive evidence demonstrates the pathophysiological importance of NOX1, the catalytic subunit of superoxide-generating enzyme NADPH oxidase, as a source of reactive oxygen species in nonphagocytic cells. However, the biochemical properties of NOX1 have not been extensively characterized due to a lack of specific immunological tools. We used a newly raised NOX1 polyclonal antibody to investigate posttranslational modifications of NOX1 overexpressed in cultured cells and in the colon, where endogenous NOX1 is highly expressed. Immunoblots of lysates from cells expressing NOX1 revealed a doublet of 56 and 60 kDa accompanied by a broad band of 60–90 kDa. Based on differential sensitivity to glycosidases, the doublet was identified as two high-mannose-type glycoforms of NOX1, whereas the broad band represented NOX1 with complex-type N-linked oligosaccharides. Deglycosylated NOX1 migrated at ~53 kDa and N-glycosylation was demonstrated in NOX1 derived from both rat and human. Site-directed mutagenesis identified N-glycosylation sites at Asn161 and Asn241 on the extracellular loop of mouse NOX1. Elimination of N-glycosylation on NOX1 did not affect its electron transferase activity, protein stability, targeting to the cell surface, or localization in F-actin-positive membrane protrusions. Taken together, these data identify the two specific sites of N-linked glycosylation of murine NOX1 and demonstrate that they are not required for normal enzyme activity, protein stability, and membrane trafficking. As is true for NOX2, the contribution of glycosylation in NOX1 to its biologic function(s) merits further study.
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NOX1 nadph oxidase is involved in endotoxin induced cardiomyocyte apoptosis
Free Radical Biology and Medicine, 2012Co-Authors: Kuniharu Matsuno, Misaki Matsumoto, Jia Zhang, Masato Katsuyama, Kazumi Iwata, Ayumi Murata, Hideo Nakamura, Kanako Ikami, Satoaki Matoba, Shinji TakaiAbstract:Abstract The functional significance of NOX1/NADPH oxidase in the heart has not been explored due to its low expression relative to other NOX homologs identified so far. We aimed to clarify the role of NOX1/NADPH oxidase in the septic heart by utilizing mice deficient in the NOX1 gene (NOX1 −/Y ). Sepsis was induced by intraperitoneal administration of lipopolysaccharides (LPS: 25 mg/kg) or cecal ligation and puncture (CLP) surgery. A marked elevation of NOX1 mRNA was demonstrated in cardiac tissue, which was accompanied by increased production of reactive oxygen species (ROS). In NOX1 −/Y treated with LPS, cardiac dysfunction and survival were significantly improved compared with wild-type mice (NOX1 +/Y ) treated with LPS. Concomitantly, LPS-induced cardiomyocyte apoptosis and activation of caspase-3 were alleviated in NOX1 −/Y . The level of phosphorylated Akt in cardiac tissue was significantly lowered in NOX1 +/Y but not in NOX1 −/Y treated with LPS or that underwent CLP surgery. Increased oxidation of cysteine residues of Akt and enhanced interaction of Akt with protein phosphatase 2A (PP2A), a major phosphatase implicated in the dephosphorylation of Akt, were demonstrated in LPS-treated NOX1 +/Y . These responses to LPS were significantly attenuated in NOX1 −/Y . Taken together, ROS derived from NOX1/NADPH oxidase play a pivotal role in endotoxin-induced cardiomyocyte apoptosis by increasing oxidation of Akt and subsequent dephosphorylation by PP2A. Marked up-regulation of NOX1 may affect the risk of mortality under systemic inflammatory conditions.
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NOX1/NADPH oxidase is involved in endotoxin-induced cardiomyocyte apoptosis
Free Radical Biology and Medicine, 2012Co-Authors: Kuniharu Matsuno, Misaki Matsumoto, Masato Katsuyama, Kazumi Iwata, Ayumi Murata, Hideo Nakamura, Kanako Ikami, Jia ZhangAbstract:Abstract The functional significance of NOX1/NADPH oxidase in the heart has not been explored due to its low expression relative to other NOX homologs identified so far. We aimed to clarify the role of NOX1/NADPH oxidase in the septic heart by utilizing mice deficient in the NOX1 gene (NOX1 −/Y ). Sepsis was induced by intraperitoneal administration of lipopolysaccharides (LPS: 25 mg/kg) or cecal ligation and puncture (CLP) surgery. A marked elevation of NOX1 mRNA was demonstrated in cardiac tissue, which was accompanied by increased production of reactive oxygen species (ROS). In NOX1 −/Y treated with LPS, cardiac dysfunction and survival were significantly improved compared with wild-type mice (NOX1 +/Y ) treated with LPS. Concomitantly, LPS-induced cardiomyocyte apoptosis and activation of caspase-3 were alleviated in NOX1 −/Y . The level of phosphorylated Akt in cardiac tissue was significantly lowered in NOX1 +/Y but not in NOX1 −/Y treated with LPS or that underwent CLP surgery. Increased oxidation of cysteine residues of Akt and enhanced interaction of Akt with protein phosphatase 2A (PP2A), a major phosphatase implicated in the dephosphorylation of Akt, were demonstrated in LPS-treated NOX1 +/Y . These responses to LPS were significantly attenuated in NOX1 −/Y . Taken together, ROS derived from NOX1/NADPH oxidase play a pivotal role in endotoxin-induced cardiomyocyte apoptosis by increasing oxidation of Akt and subsequent dephosphorylation by PP2A. Marked up-regulation of NOX1 may affect the risk of mortality under systemic inflammatory conditions.
