Ethanol‐Induced Modulation of Inducible Nitric‐Oxide Synthase Activity in Human A172 Astrocytoma Cells

Davis, Randall L.; Dertien, Janet; Syapin, Peter J.

doi:10.1111/j.1530-0277.2002.tb02685.x

Cited by 38 publications

(12 citation statements)

References 46 publications

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“…Details on the time course of cytokine-induced iNOS expression in A172 cells, at the level of mRNA and protein, have been discussed in a previous report (Davis and Syapin 2004). Likewise, cytokine-stimulated iNOS activity in A172 cells as measured by nitrite accumulation and the citrulline conversion assay has been characterized by our group (Davis et al 2002;Davis and Syapin 2004).…”

Section: Discussionmentioning

confidence: 94%

“…A172 cells were maintained as previously described (Davis et al 2002), and experimental cultures were seeded at a cell density (1×10 4 /cm 2 ) to provide 80-90% confluence at the time of treatment.…”

Section: Cell Culturementioning

confidence: 99%

“…Optimal induction of iNOS expression in A172 cells requires co-exposure to a mixture of cytokines (Davis et al 2002). Thus, iNOS was induced by 8-24 h exposure to a cytokine mixture containing 100 ng/ml human recombinant IFNγ (Peprotech Inc., Rocky Hill, NJ), 30 ng/ml human recombinant TNFα (Peprotech), and 5 ng/ml human recombinant IL-1β (Peprotech).…”

Section: Inos Inductionmentioning

confidence: 99%

“…To quantitate iNOS activity, nitrite accumulation in the culture media was determined spectrophotometrically 24 h post-stimulation using the Griess reagent as previously described (Davis et al 2002). Quantitative analysis of NOS2 gene expression was evaluated using the quantitative real-time polymerase chain reaction (qRT-PCR) and the SYBR green I reporter assay.…”

Section: Inos Inductionmentioning

confidence: 99%

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β-Funaltrexamine Inhibits Inducible Nitric-oxide Synthase Expression in Human Astroglial Cells

Davis

Buck

Saffarian

et al. 2008

J Neuroimmune Pharmacol

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The inducible isoform of nitric-oxide synthase (iNOS) is involved in neuropathogenesis associated with infection and disease in the brain. Hence, there is considerable interest in the identification of therapeutic interventions to prevent iNOS-mediated pathology. Astroglia are a major site of iNOS expression during neuropathogenesis. To mimic a key component of neuroinflammation, human A172 astroglial cells were exposed in vitro to a cytokine mixture containing interferon γ, tumor necrosis factor α, and interleukin-1β, resulting in significant iNOS expression. Next, we assessed the effects of the mu opioid receptor antagonist, β-funaltrexamine (β-FNA), on cytokine induced iNOS expression in human astroglia. β-FNA dose-dependently inhibited iNOS expression. β-FNA transcriptionally (or pre-transcriptionally) inhibited cytokine-induced iNOS activation as indicated by a significant decrease in NOS2 messenger RNA expression. Further characterization of the novel, anti-inflammatory actions of β-FNA may provide insights for pharmacologic strategies to treat or prevent brain pathologies associated with neuroinflammation.

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Section: Discussionmentioning

confidence: 94%

Section: Cell Culturementioning

confidence: 99%

Section: Inos Inductionmentioning

confidence: 99%

Section: Inos Inductionmentioning

confidence: 99%

See 2 more Smart Citations

β-Funaltrexamine Inhibits Inducible Nitric-oxide Synthase Expression in Human Astroglial Cells

Davis

Buck

Saffarian

et al. 2008

J Neuroimmune Pharmacol

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“…Additionally, we have previously characterized proinflammatory pathways in A172 cells (Davis et al, 2002;Davis and Syapin, 2004a;Davis and Syapin, 2004b). A172 cells were maintained in Dulbecco's modified Eagle's medium (DMEM) containing 2 mM L-glutamine, 10% fetal bovine serum, 1% nonessential amino acids, 50 U/ml penicillin, 0.05 mg/ml streptomycin and 2 g/ml amphotericin B. Cultures were maintained in a humidified incubator at 37°C, 5% CO 2 and 95% air and culture medium changed every 48-72 h. Experimental cultures were seeded at a cell density (1 × 10 4 /cm 2 ) in order to provide 80-90% confluence at the time of treatment.…”

Section: Cell Culturementioning

confidence: 99%

The opioid antagonist, β-funaltrexamine, inhibits chemokine expression in human astroglial cells

Davis

Buck

Saffarian

et al. 2007

Journal of Neuroimmunology

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Emerging evidence indicates that neuroinflammatory responses in astroglia, including chemokine expression, are altered by opioids. Astroglial chemokines, such as CXCL10, are instrumental in response to many neuropathological insults. Opioid mediated disruption of astroglial CXCL10 expression may be detrimental in opioid abusers or patients receiving acute opioid therapy. We have characterized the in vitro effects of opioids on CXCL10 protein expression in human astroglial (A172) cells. The proinflammatory cytokine, tumor necrosis factor (TNF)α induced CXCL10 expression in A172 cells. Using MG-132, helenalin and SN50 [inhibitors of the transcription factor, nuclear factor (NF)-κB], we determined that NF-κB activation is instrumental in TNFα induced CXCL10 expression in A172 astroglia. Morphine exposure during the 24 h TNFα stimulation period did not alter CXCL10 expression. However, fentanyl, a more potent mu opioid receptor (MOR) agonist, inhibited TNFα induced CXCL10 expression. Interestingly, neither the nonselective opioid receptor antagonist, naltrexone nor β-funaltrexamine (β-FNA), a highly selective MOR antagonist, blocked fentanyl mediated inhibition of TNFα induced CXCL10 expression. Rather, β-FNA dose dependently inhibited TNFα induced CXCL10 expression with a greater potency than that observed for fentanyl. Immunoblot analysis indicated that morphine, fentanyl and β-FNA each reduced TNFα induced nuclear translocation of NF-κB p65. These data show that β-FNA and fentanyl inhibit TNFα induced CXCL10 expression via a MOR independent mechanism. Data also suggest that inhibition of TNFα induced CXCL10 expression by fentanyl and β-FNA is not directly related to a reduction in NF-κB p65 nuclear translocation. Further investigation is necessary in order to fully elucidate the mechanism through which these two opioid compounds inhibit CXCL10 expression. Understanding the mechanism by which chemokine expression is suppressed, particularly by the opioid antagonist, β-FNA, may provide insights into the development of safe and effective treatments for neuroinflammation.

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Severe alcohol‐induced neuronal deficits in the hippocampus and neocortex of neonatal mice genetically deficient for neuronal nitric oxide synthase (nNOS)

Bonthius

McKim

Koele

et al. 2006

J of Comparative Neurology

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Alcohol can severely damage the developing brain, and neuronal loss is a critical component of this injury. Thus, identification of molecular factors that ameliorate alcohol-induced neuronal loss is of great importance. Previous in vitro work has demonstrated that nitric oxide (NO) protects neurons against alcohol toxicity. We tested the hypothesis that neonatal mice carrying a null mutation for neuronal nitric oxide synthase (nNOS), the enzyme that synthesizes NO in neurons, have an increased vulnerability to alcohol-induced neuronal loss in the neocortex and hippocampus. Wildtype mice and nNOS-/- mice received ethanol (0.0, 2.2, 3.3, or 4.4 g/kg) daily over postnatal days (P) 4-9 and were sacrificed on P10. The number of hippocampal CA1 and CA3 pyramidal cells, dentate gyrus granule cells, and neocortical neurons were determined using stereological methods. Alcohol pharmacokinetics did not differ between wildtype and nNOS-/- strains. Alcohol induced dose-dependent reductions in all four neuronal populations, and the losses were substantially more severe in the nNOS-/- mice than in wildtype. Furthermore, the threshold dose of alcohol to induce cell death was lower in the nNOS-/- mice than in the wildtype mice for all neuronal populations. While nNOS deficiency worsened alcohol-induced neuronal losses, the magnitude of this exacerbation varied among brain regions and depended on alcohol dose. These results demonstrate that nNOS deficiency decreases the ability of developing neurons in vivo to survive the toxic effects of alcohol and strengthen the hypothesis that NO exerts a neuroprotective effect against alcohol toxicity in the developing brain.

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Ethanol‐Induced Modulation of Inducible Nitric‐Oxide Synthase Activity in Human A172 Astrocytoma Cells

Cited by 38 publications

References 46 publications

β-Funaltrexamine Inhibits Inducible Nitric-oxide Synthase Expression in Human Astroglial Cells

β-Funaltrexamine Inhibits Inducible Nitric-oxide Synthase Expression in Human Astroglial Cells

The opioid antagonist, β-funaltrexamine, inhibits chemokine expression in human astroglial cells

Severe alcohol‐induced neuronal deficits in the hippocampus and neocortex of neonatal mice genetically deficient for neuronal nitric oxide synthase (nNOS)

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