inducible nitric oxide synthase inos inhibitor 1400 w  (MedChemExpress)

Journal: International journal of molecular sciences

Article Title: Treatment of Status Epilepticus after Traumatic Brain Injury Using an Antiseizure Drug Combined with a Tissue Recovery Enhancer Revealed by Systems Biology.

doi: 10.3390/ijms241814049

Figure Lengend Snippet: Figure 2. Effect of in silico-identified drugs on tissue biomarkers of neuroprotection, oxidative stress, and neuroinflammation. (A) Neuronal viability. For a comparative viability analysis, neu- ronal survival in LPS/IFNγ+-treated neuronal–microglial co-cultures was set to 0% and that in co-cultures treated with iNOS inhibitor 1400 W (positive treatment control) to 100%. Trichostatin A at a 50 nM concentration increased neuronal survival from 0% to 73% (p < 0.001 compared with LPS/IFNγ+). The neuroprotective effect of TSA was almost as good as that of the positive control 1400 W (73% vs. 100%, p < 0.001). Also, chlorpromazine at a 50-µM concentration increased neuronal survival from 0% to 57% (p < 0.001) and calpain inhibitor at a 50-µM concentration increased neu- ronal survival from 0% to 47% (p < 0.001). Geldanamycin and tranylcypromine showed no consistent neuroprotection. IL10 had no effect on neuronal viability. Note that in untreated co-cultures without neuroinflammation (LPS/IFNγ−), neuronal viability was 38% of that in untreated BV2 neuronal cultures (p < 0.001) and 48% of that in 1400 W treated inflammation-exposed co-cultures (p < 0.001). Wells with cortical neurons only (BV2−cells without LPS/IFNγ+ exposure) showed the greatest viability. (B) Nitrite levels. For a comparative viability analysis, nitrite levels in LPS/IFNγ+-treated neuronal–microglial co-culture medium were set to 100% and that in co-culture medium treated with iNOS inhibitor 1400 W (positive treatment control) was set to 0%. Trichostatin A at a 50 nM concentration reduced the LPS/IFNγ+-induced nitrite release from 100% to 28% (p < 0.001). At a 10 nM concentration, there was a slight increase in nitrite levels to 121% (p < 0.01). Calpain inhibitor at 50 µM reduced nitrite release from 100% to −21% (p < 0.001). Chlorpromazine at 50 µM reduced nitrite levels to 81% (p < 0.001). At 10 µM, tranylcypromine reduced nitrite levels from 100% to 89% (p < 0.05) and at 1 µM, to 81% (p < 0.001). Interestingly, tranylcypromine at 100 µM had no effect (p > 0.05). Geldanamycin at 100 nM concentrations resulted in low nitrite levels, likely due to the cell death that was visualized in microscopic images (data not shown). IL10 had a mild effect on the LPS/IFNγ+-induced increase in nitrite levels, reducing it to 63% (p < 0.001). Note that neuron-only (BV2−) and LPS/IFNγ−neuronal–microglial co-cultures were not exposed to neuroinflammation, and thus showed no nitrite release to the culture medium. (C) TNFα levels. For a comparative viabil- ity analysis, TNFα levels in LPS/IFNγ+-treated neuronal–microglial co-culture medium were set to 100% and that in co-culture medium treated with TNFα inhibitor, IL10 (positive treatment control) to 0%. Trichostatin A at 10 nM concentration reduced TNFα levels from 100% to −2% (p < 0.001) and at 50 nM to −139% (p < 0.001), being better than the positive control. Calpain inhibitor at 50 µM reduced TNFα levels from 100% to −226% (p < 0.001). Chlorpromazine at 50 µM reduced TNFα levels from 100% to −21% (p < 0.001). The iNOS inhibitor, at 1400 W, reduced TNFα levels from 100% to 61% (p < 0.05). Abbreviations: BV2−, cortical neuronal cultures without BV2-microglial cells; IL10+, co-cultures treated with positive control anti-inflammatory incytokine, interleukin 10; LPS/IFNγ−, co-culture wells without lipopolysaccharides and interferon γ-induced neuroinflam- mation; LPS/IFNγ+, wells with lipopolysaccharides and interferon γ-induced neuroinflammation (untreated controls); TNFα, tumor necrosis factor α; 1400 W+, co-cultures treated with positive con- trol, inducible nitric oxide synthase (iNOS) inhibitor, 1400 W hydrochloride. Statistical significance: ***, p < 0.001; **, p < 0.01; *, p < 0.05 compared with LPS + IFNγ+ (linear regression in R). The number of independent experiments in panel A was three and in panels B-C was four. Data are presented as box plots with whiskers (minimum and maximum; box: interquartile range; line: median). Each dot shows an experimental value.

Article Snippet: Anti-inflammatory cytokine, interleukin 10 (IL10, 50 μg/mL, #500-M128, Peprotech, Rocky Hill, NJ, USA; final concentration of 50 ng/mL), and the neuroprotective inducible nitric oxide synthase (iNOS) inhibitor 1400 W hydrochloride (2 mM, #1415, Tocris, Bristol, UK; final concentration of 20 μM) were used as positive controls.

Techniques: In Silico, Control, Concentration Assay, Positive Control, Co-Culture Assay

Journal: PLoS ONE

Article Title: A Key Role for the Endothelium in NOD1 Mediated Vascular Inflammation: Comparison to TLR4 Responses

doi: 10.1371/journal.pone.0042386

Figure Lengend Snippet: Aortic rings with endothelium intact (+EC) and denuded (−EC) were treated for 24 hours with media alone (CTRL), C12-iE-DAP 1 µg/ml (NOD1 agonist) or LPS 1 µg/ml (TLR4 agonist) (A) In some experiments, cells were treated for a further 24 hour period with replacement of media and agonists (48 hours; B). In addition, endothelium intact rings were pre-treated with the iNOS inhibitor 1400 W 1 µM (C) or the IκBβ inhibitor SC-514 30 µM (D) for 30 minutes prior to 24 hours treatment with agonists. iNOS activity was assessed by measurement of nitrite (breakdown product of nitric oxide) by the Griess assay. Results are expressed as mean ± SEM for n = 9 (A+B) or n = 4 (C+D) animals. Data was analysed by one-way ANOVA followed by Bonferroni’s Multiple Comparison Test (*P<0.05) or by two-way ANOVA followed by Bonferroni’s post tests (+P<0.05).

Article Snippet: The inhibitor of NFκB kinase subunit beta (IκBβ) inhibitor SC-514, transforming growth factor β activating kinase (TAK1) inhibitor 5Z-7-oxozeaonol and the pan-caspase inhibitor Z-VAD-fmk were from Tocris Bioscience (UK) and the inducible nitric oxide synthase (iNOS) inhibitor 1400 W from Cayman Chemical (USA).

Techniques: Activity Assay, Griess Assay