Differential Modulation of NF-κB in Neurons and Astrocytes Underlies Neuroprotection and Antigliosis Activity of Natural Antioxidant Molecules

Figure 2

Effect of antioxidants on ROS levels and astrocyte viability. (a) Survival by the MTT assay of cortical astrocytes stimulated with TNFα (10 ng/ml) or LPS (1 μg/ml) for 24 h and the effect of RSV (10 μM), QRC (10 μM), ALA (10 μM), CRC (10 μM), LYC (10 μM), OLP (100 μg/ml), GTE (12.5 μg/ml), NAC (300 μM), or pool. (b) Quantitation of astrocytic ROS levels by FACS analysis of DCH2F-DA fluorescence following stimulation with TNFα (10 ng/ml) for 6 h, or LPS (1 μg/ml) for 12 h, and effect of the indicated antioxidants and pool. (c) MTT assay on astrocytes exposed to H₂O₂ (200 μM) for 24 h in the presence or absence of the indicated antioxidants and pool. (d) ROS levels in astrocytes treated with H₂O₂ (200 μM) for 6 h in the presence or absence of the indicated antioxidants and pool. All data are expressed as percent of CTR. MTT data are the of three independent experiments, each performed with 4-6 samples for each treatment. ROS data are the of three independent experiments, each performed in duplicate. (e, f) Western blot analysis and quantitation of vGLUT levels in astrocytes treated for 24 h with TNFα (10 ng/ml) or LPS (1 μg/ml) and the effect of RSV cotreatment. Data, expressed as percent of CTR, are the of three independent experiments. , , and versus CTR (ANOVA and Dunnett’s multiple comparison test).

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