Phenolic Compounds of Red Wine Aglianico del Vulture Modulate the Functional Activity of Macrophages via Inhibition of NF-κB and the Citrate Pathway

Figure 4

RWP inhibited NF-κB transcription factor. (a) Primary human monocytes were incubated with RWP 20 μg/mL or RWP 200 μg/mL and activated to macrophages with 1 μg/mL LPS. Specific antibodies detected the expression levels of subunit p65 of NF-κB and β-actin. The intensities of immunolabeled protein bands were measured by a quantitative software and normalized to β-actin; values obtained are reported under western blot images. Protein expression levels in control sample were taken as 1, and other samples were expressed in proportion to the control. (b) HEK293 cells were transfected with NF-κB luciferase reporter plasmid and treated with LPS in the presence or not of RWP 20 μg/mL or RWP 200 μg/mL. Bar chart reports the of three independent experiments, each in triplicate. According to one-way ANOVA, differences were significant (). Therefore, Tukey’s post hoc test was performed, and different letters indicate significant differences between treatments at . (c-d) Immunocytochemistry experiments were performed to identify the cellular localization of subunit p65 of NF-κB, recognized by a specific antibody. Cells were treated with RWP 20 μg/mL or RWP 200 μg/mL and activated with LPS. C: control; L: LPS; R20: RWP 20 μg/mL; R200: RWP 200 μg/mL.

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