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

Luengo-Blanco, Marcos; Prando, Carolina; Bustamante, Jacinta; Aragão-Filho, Walmir Cutrim; Pereira, Pedro Paulo; Rehder, Jussara; Padden, Carolyn; Casanova, Jean‐Laurent; Newburger, Peter E.; Condino‐Neto, Antônio

doi:10.1182/blood-2007-07-099267

Cited by 29 publications

(30 citation statements)

References 46 publications

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“…Among other mechanisms, activation of NADPH oxidase (NOX) by NF-κB represents an important factor in OS induction, as NOX is one of the most important sources of superoxide generation (Manea et al, 2007; Luengo-Blanco et al . 2008).…”

Section: Discussionmentioning

confidence: 99%

NF-κB in the mechanism of brain edema in acute liver failure: Studies in transgenic mice

Jayakumar

Bethea

Tong

et al. 2011

Neurobiology of Disease

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Astrocyte swelling and brain edema are major complications of the acute form of hepatic encephalopathy (acute liver failure, ALF). While elevated brain ammonia level is a well-known etiological factor in ALF, the mechanism by which ammonia brings about astrocyte swelling is not well understood. We recently found that astrocyte cultures exposed to ammonia activated nuclear factor-kappaB (NF-κB), and that pharmacological inhibition of such activation led to a reduction in astrocyte swelling. Although these findings suggest the involvement of NF-κB in astrocyte swelling in vitro, it is not known whether NF-κB contributes to the development of brain edema in ALF in vivo. Furthermore, pharmacological agents used to inhibit NF-κB may have non-specific effects. Accordingly, we used transgenic (Tg) mice that have a functional inactivation of astrocytic NF-κB and examined whether these mice are resistant to ALF-associated brain edema. ALF was induced in mice by treatment with the hepatotoxin thioacetamide (TAA). Wild type (WT) mice treated with TAA showed a significant increase in brain water content (1.65%) along with prominent astrocyte swelling and spongiosis of the neuropil, consistent with the presence of cytotoxic edema. These changes were not observed in Tg mice treated with TAA. Additionally, WT mice with ALF showed an increase in inducible nitric oxide synthase (iNOS) immunoreactivity in astrocytes from WT mice treated with TAA (iNOS is known to be activated by NF-κB and to contribute to cell swelling). By contrast, Tg mice treated with TAA did not exhibit brain edema, histological changes nor an increase in iNOS immunoreactivity. We also examined astrocytes cultures derived from Tg mice to determine whether these cells exhibit a lesser degree of swelling and cytopathological changes following exposure to ammonia. Astrocyte cultures derived from Tg mice showed no cell swelling nor morphological abnormalities when exposed to ammonia for 24 h. By contrast, ammonia significantly increased cell swelling (31.7%) in cultured astrocytes from WT mice and displayed cytological abnormalities. Moreover, we observed a lesser increment in inducible nitric oxide synthase and NADPH oxidase activity (both are also known to be activated by NF-κB and to contribute to astrocyte swelling) in astrocyte cultures from Tg mice treated with ammonia, as compared to ammonia-treated WT mice astrocytes. These findings strongly suggest that activation of NF-κB is a critical factor in the development of astrocyte swelling/brain edema in ALF.

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Section: Discussionmentioning

confidence: 99%

NF-κB in the mechanism of brain edema in acute liver failure: Studies in transgenic mice

Jayakumar

Bethea

Tong

et al. 2011

Neurobiology of Disease

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“…Reverse transcription was performed on 3 μg of RNA with SuperScript II RT (Life Technologies) from random hexamers. The cDNA was amplified by 30 cycles of PCR with oligonucleotides specific for gp91 -phox (Gene Bank NM 00397) or β-actin (Gene Bank NM 001101) as previously described [Luengo-Blanco et al, 2008]. PCR products were analyzed by agarose electrophoresis and ethidium bromide staining, and quantitated by digital photograph and computer analysis with Molecular Analyst software (Bio-Rad).…”

Section: Methodsmentioning

confidence: 99%

“…In vivo, mutations in the IKBKG gene, which encodes the NF-κB regulatory protein NEMO, cause anhidrotic ectodermal dysplasia associated with immunodeficiency (EDA-ID; OMIM 300291), a genetic disorder characterized by the aberrant development of skin appendages, including eccrine sweat glands (anhidrosis), as well as severe infections caused by mycobacteria and other microorganisms [Bustamante et al, 2011]. In vitro, EBV-transformed B cells from EDA-ID patients, as well as U937 cells stably transfected with an NF-κB repressor (IκBα-S32A/S36A), showed significantly decreased superoxide release and CYBB gene expression [Luengo-Blanco et al, 2008]. Gallin’s group has also shown impaired superoxide production in neutrophils from patients with NEMO deficiency [Singh et al, 2009].…”

mentioning

confidence: 99%

Regulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF‐κB Binding Site

Frazão

Thain

Zhu

et al. 2015

J of Cellular Biochemistry

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The human CYBB gene encodes the gp91-phox component of the phagocyte oxidase enzyme complex, which is responsible for generating superoxide and other downstream reactive oxygen species essential to microbial killing. In the present study, we have identified by sequence analysis a putative NF-κB binding site in a DNase I hypersensitive site, termed HS-II, located in the distant 5′ flanking region of the CYBB gene. Electrophoretic mobility assays showed binding of the sequence element by recombinant NF-κB protein p50 and by proteins in nuclear extract from the HL-60 myeloid leukemia cell line corresponding to p50 and to p50/p65 heterodimers. Chromatin immunoprecipitation demonstrated NF-κB binding to the site in intact HL-60 cells. Chromosome conformation capture (3C) assays demonstrated physical interaction between the NF-κB binding site and the CYBB promoter region. Inhibition of NF-κB activity by salicylate reduced CYBB expression in peripheral blood neutrophils and differentiated U937 monocytic leukemia cells. U937 cells transfected with a mutant inhibitor of κB “super-repressor” showed markedly diminished CYBB expression. Luciferase reporter analysis of the NF-κB site linked to the CYBB 5′ flanking promoter region revealed enhanced expression, augmented by treatment with interferon-γ. These studies indicate a role for this distant, 15 kb upstream, binding site in NF-κB regulation of the CYBB gene, an essential component of phagocyte-mediated host defense.

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“…One important consequence of NF-jB activation is the stimulation of NOX, 33,34 and such activation of NOX is known to mediate astrocyte swelling after hyperammonemia. 35,36 We therefore examined whether trauma-induced NF-jB activation contributes to NOX activation.…”

Section: Nf-jb Inhibition Diminishes the Trauma-induced Nox Activationmentioning

confidence: 99%

“…6 Activation of NOX by NF-jB represents an important factor in the induction of OS, as NOX is one of the most important sources of superoxide formation. 33,34 Activation of NOX has been shown to stimulate cell swelling in cultured astrocytes after ammonia treatment. 36 Consistent with these observations, we found increased NOX activity after trauma in cultured astrocytes and that BAY 11-7082, an inhibitor of NF-jB, significantly diminished such activation.…”

Section: Nf-jb In Trauma-induced Astrocyte Swellingmentioning

confidence: 99%

Activation of NF-κB Mediates Astrocyte Swelling and Brain Edema in Traumatic Brain Injury

Jayakumar

Tong

Ruíz-Cordero

et al. 2014

Journal of Neurotrauma

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Brain edema and associated increased intracranial pressure are major consequences of traumatic brain injury (TBI). While astrocyte swelling (cytotoxic edema) represents a major component of the brain edema in the early phase of TBI, its mechanisms are unclear. One factor known to be activated by trauma is nuclear factor-jB (NF-jB). Because this factor has been implicated in the mechanism of cell swelling/brain edema in other neurological conditions, we examined whether NF-jB might also be involved in the mediation of post-traumatic astrocyte swelling/brain edema. Here we show an increase in NF-jB activation in cultured astrocytes at 1 and 3 h after trauma (fluid percussion injury, FPI), and that BAY 11-7082, an inhibitor of NF-jB, significantly blocked the trauma-induced astrocyte swelling. Increased activities of nicotinamide adenine dinucleotide phosphate-oxidase and the Na + , K + , 2Cl -cotransporter were also observed in cultured astrocytes after trauma, and BAY 11-7082 reduced these effects. We also examined the role of NF-jB in the mechanism of cell swelling by using astrocyte cultures derived from transgenic (Tg) mice with a functional inactivation of astrocytic NF-jB. Exposure of cultured astrocytes from wild-type mice to in vitro trauma (3 h) caused a significant increase in cell swelling. By contrast, traumatized astrocyte cultures derived from NF-jB Tg mice showed no swelling. We also found increased astrocytic NF-jB activation and brain water content in rats after FPI, while BAY 11-7082 significantly reduced such effects. Our findings strongly suggest that activation of astrocytic NF-jB represents a key element in the process by which cytotoxic brain edema occurs after TBI.

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

Cited by 29 publications

References 46 publications

NF-κB in the mechanism of brain edema in acute liver failure: Studies in transgenic mice

NF-κB in the mechanism of brain edema in acute liver failure: Studies in transgenic mice

Regulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF‐κB Binding Site

Activation of NF-κB Mediates Astrocyte Swelling and Brain Edema in Traumatic Brain Injury

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