Redox-sensitive regulation of macrophage-inducible nitric oxide synthase expression in vitro does not correlate with the failure of apocynin to prevent lung inflammation induced by endotoxin

Viačková, Daniela; Pekarová, Michaela; Crhák, Tomáš; Búcsaiová, Martina; Matiašovic, Ján; Lojek, Antonı́n; Kubala, Lukáš

doi:10.1016/j.imbio.2010.09.005

Cited by 13 publications

(13 citation statements)

References 44 publications

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“…The NADPH oxidase activity was determined in cell lysates prepared according to the well-established protocol [31]. Briefly, to the 100 μ L of tested solution, lucigenin was added at final concentration 5 μ M. After that, NADPH at final concentration 100 μ M was added to start the production of O 2 • − .…”

Section: Methodsmentioning

confidence: 99%

“…Real-time PCR was performed on RTCykler7300 (Applied Biosystems), and the parameters of amplification were set up according to the supplier's instructions. The fold of the mRNA induction was calculated using the ΔΔ C t method, with GAPDH as a housekeeping gene (TaqMan Rodent GAPDH Control reagent, Applied Biosystems, USA) [31]. …”

Section: Methodsmentioning

confidence: 99%

“…Using an electroporation system (Gene Pulser II, Bio-Rad laboratopries, USA, for details see [31]), cells were transfected with plasmids containing the shRNA construct, against iNOS and negative control plasmid with a scrambled sequence (Origene, USA). Stably transfected cells were grown in DMEM + 5% FBS and 5 μ g/mL puromycin.…”

Section: Methodsmentioning

confidence: 99%

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New Role for L-Arginine in Regulation of Inducible Nitric-Oxide-Synthase-Derived Superoxide Anion Production in Raw 264.7 Macrophages

Pekarová

Lojek

Martišková

et al. 2011

The Scientific World JOURNAL

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Dietary supplementation with L-arginine was shown to improve immune responses in various inflammatory models. However, the molecular mechanisms underlying L-arginine effects on immune cells remain unrecognized. Herein, we tested the hypothesis that a limitation of L-arginine could lead to the uncoupled state of murine macrophage inducible nitric oxide synthase and, therefore, increase inducible nitric-oxide-synthase-derived superoxide anion formation. Importantly, we demonstrated that L-arginine dose- and time dependently potentiated superoxide anion production in bacterial endotoxin-stimulated macrophages, although it did not influence NADPH oxidase expression and activity. Detailed analysis of macrophage activation showed the time dependence between LPS-induced iNOS expression and increased O2∙− formation. Moreover, downregulation of macrophage iNOS expression, as well as the inhibition of iNOS activity by NOS inhibitors, unveiled an important role of this enzyme in controlling O2∙− and peroxynitrite formation during macrophage stimulation. In conclusion, our data demonstrated that simultaneous induction of NADPH oxidase, together with the iNOS enzyme, can result in the uncoupled state of iNOS resulting in the production of functionally important levels of O2∙− soon after macrophage activation with LPS. Moreover, we demonstrated, for the first time that increased concentrations of L-arginine further potentiate iNOS-dependent O2∙− formation in inflammatory macrophages.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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New Role for L-Arginine in Regulation of Inducible Nitric-Oxide-Synthase-Derived Superoxide Anion Production in Raw 264.7 Macrophages

Pekarová

Lojek

Martišková

et al. 2011

The Scientific World JOURNAL

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“…Both of these cell lines are commonly used for experiments focused on the examination of macrophage functions under resting and activated conditions (Pekarova et al, 2009a(Pekarova et al, , 2009bViackova et al, 2010). Moreover, the difference in their origin could contribute to the amount of information about the potential universal effects of methylarginines in regulation of macrophage activation.…”

Section: Cell Culturesmentioning

confidence: 99%

“…The luciferase reporter construct pBIIX-LUC (kindly donated by Dr. Kalle Saksela, Institute of Molecular Medicine, University of Tampere, Finland) was transiently transfected into RAW 264.7 and MHS macrophages using an electroporation system (Gene Pulser II, Bio-Rad Laboratories) (Viackova et al, 2010). The construction of pfLUC (from pBL-KS plasmid) and NF-kB-driven plasmid (pBIIX-LUC) was described in details elsewhere (Saksela and Baltimore, 1993).…”

Section: Nf-κb Binding Activitymentioning

confidence: 99%

Asymmetric dimethylarginine regulates the lipopolysaccharide-induced nitric oxide production in macrophages by suppressing the activation of NF-kappaB and iNOS expression

Pekarová

Kubala

Martišková

et al. 2013

European Journal of Pharmacology

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Apocynin inhibits Toll-like receptor-4-mediated activation of NF-κB by suppressing the Akt and mTOR pathways

Nam

Kim

Sohn

et al. 2016

Naunyn-Schmiedeberg's Arch Pharmacol

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Microbial product lipopolysaccharide has been shown to be involved in the pathogenesis of inflammatory skin diseases. Apocynin has demonstrated to have an anti-inflammatory effect. However, the effect of apocynin on the Toll-like receptor-4-dependent activation of Akt, mammalian target of rapamycin (mTOR), and nuclear factor (NF)-κB pathway, which is involved in productions of inflammatory mediators in keratinocytes, has not been studied. Using human keratinocytes, we investigated the effect of apocynin on the inflammatory mediator production in relation to the Toll-like receptor-4-mediated-Akt/mTOR and NF-κB pathways, which regulates the transcription genes involved in immune and inflammatory responses. Apocynin, Akt inhibitor SH-5, Bay 11-7085 and N-acetylcysteine each attenuated the lipopolysaccharide-induced production of cytokines, PGE, and chemokines, changes in the levels of Toll-like receptor-4, p-Akt, mTOR, and NF-κB, and production of reactive oxygen species in keratinocytes. The results show that apocynin appears to attenuate the lipopolysaccharide-stimulated production of inflammatory mediators in keratinocytes by suppressing the Toll-like receptor-4-mediated activation of the Akt, mTOR, and NF-κB pathways. The effect of apocynin appears to be attributed to its inhibitory effect on the production of reactive oxygen species. Apocynin appears to attenuate the microbial product-mediated inflammatory skin diseases.

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Redox-sensitive regulation of macrophage-inducible nitric oxide synthase expression in vitro does not correlate with the failure of apocynin to prevent lung inflammation induced by endotoxin

Cited by 13 publications

References 44 publications

New Role for L-Arginine in Regulation of Inducible Nitric-Oxide-Synthase-Derived Superoxide Anion Production in Raw 264.7 Macrophages

New Role for L-Arginine in Regulation of Inducible Nitric-Oxide-Synthase-Derived Superoxide Anion Production in Raw 264.7 Macrophages

Asymmetric dimethylarginine regulates the lipopolysaccharide-induced nitric oxide production in macrophages by suppressing the activation of NF-kappaB and iNOS expression

Apocynin inhibits Toll-like receptor-4-mediated activation of NF-κB by suppressing the Akt and mTOR pathways

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