Anti-Inflammatory Effect of Procyanidins from Wild Grape (Vitis amurensis) Seeds in LPS-Induced RAW 264.7 Cells
Effects of WGP on LPS-induced p65 nuclear translocation and p65 and IκBα phosphorylation in RAW 264.7 cells. (a) The cells were treated with 1 μg/mL LPS alone or with WGP (35 μg/mL) for 2 h. Cytosolic and nuclear fractions were prepared and analyzed by Western blotting. The ratio of immunointensity between the p65 (nuclear fraction: NE)/p65 (cytosolic fraction: CE) and the Lamin B and β-actin was calculated. Each bar (open bar, p65 (NE); closed bar, p65 (CE)) represents means ± S.D. from three independent experiments. (b) Cells were pretreated with WGP (35 μg/mL) for 1 h followed by stimulation with LPS for 2 h. Samples were stained by anti-p65 antibody and DAPI then prepared for confocal microscopy analysis. (c) The whole cells were treated with WGP at 35 μg/mL for 2 h and treated with LPS for 1 h. Equal amounts of total protein were subjected to Western blot analysis. The ratio of immunointensity between the p-p65/p-IκBα and the β-actin was calculated. Each bar (open bar, p-p65; closed bar, p-IκBα) represents means ± S.D. from three independent experiments. indicates differences from the unstimulated control group. indicates differences from the LPS-treated group.