Figure 3.

Effect of 100 µm Agm on O₂⁻ level, XO activity, and lipid peroxidation in control and BV-2 cells stimulated with Lps. BV-2 cells were pretreated with Agm (100 µm) for 30 min and then stimulated with Lps (1 µg/mL) for an additional 4 and 6 h (qPCR analysis) or 24 h (O₂⁻, XO, 4-HNE, and MDA). (a) NOX2 gene (Cybb) expression relative to the expression of the Gapdh gene. Superoxide-producing (b) and XO activity (c). Panel (d) represents immunofluorescence labeling against 4-HNE bound proteins (red) counterstained with Hoechst (blue); plotted scale bar applies to all micrographs. (e) Mean per-pixel fluorescence intensity (MFI) of 4-HNE per cell was measured in ~200 cells per experimental group and analyzed as described in Materials and Methods. (f) The intracellular concentration of MDA. Data collected from all assays are presented as mean ± SEM, from three separate determinations, and were analyzed by Kruskal–Wallis test followed by Dunn’s multiple comparisons tests. * p < 0.05 compared with a control group; # compared with Lps group.