Norfluoxetine Prevents Degeneration of Dopamine Neurons by Inhibiting Microglia-Derived Oxidative Stress in an MPTP Mouse Model of Parkinson’s Disease

Figure 4

Norfluoxetine attenuates LPS-induced activation of NADPH oxidase and NO production in cortical microglial cultures. Cortical cultures of rat microglia were pretreated with fluoxetine (50 and 100 μM) or norfluoxetine (50 μM) for 6 hr followed by exposure to LPS (100 μg/ml) or vehicle as a control or for 24 h. Cultured cells were homogenized, fractionated, and analyzed NADPH oxidase by Western blot analysis (a, b), and media from cultures were collected for measuring the levels of nitrate (c). (a, b) Western blot analysis showing p47^phox activation. The levels of p47^phox (NADPH oxidase subunits) in the cytosol and membrane were determined by normalizing calnexin as membrane protein level. CON: control; NF: norfluoxetine; L: LPS; L + NF: LPS and norfluoxetine. Values are expressed as a percentage of the total level of p47^phox. , compared with nontreated control; , compared with LPS only treated. (c) Nitrate levels are presented as means ± SEM of triplicate cultures on two separate plates. , compared with nontreated control cultures; , , and , compared with LPS-only-treated cultures (ANOVA and Student–Newman–Keuls analyses).

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