Myricitrin Modulates NADPH Oxidase-Dependent ROS Production to Inhibit Endotoxin-Mediated Inflammation by Blocking the JAK/STAT1 and NOX2/p47^phox Pathways

Figure 8

p47^phox and gp91^phox RNAi inhibited LPS-induced JAK/STAT1 activation and decreased iNOS and COX-2 expression. RAW264.7 cells were, respectively, transfected with shRNA targeting p47^phox or gp91^phox and cotransfected with p47^phox and gp91^phox shRNA. The scrambled RNAi group was transfected with pFU-GW plasmid vector. Forty-eight hours after transfection, RAW264.7 cells were stimulated with LPS (100 ng/ml) for 15 min (a), 4 h (b), and 16 h (c). Cell lysates were prepared and subjected to western blotting by using phospho-JAK1 and phospho-JAK2 antibodies (a), phospho-STAT1 antibody (b), and anti-iNOS and anti-COX-2 antibodies (c). GAPDH and nonphosphorylated antibodies were estimated as the protein-loading control for each lane. p47^phox and gp91^phox were estimated to identify the effect of interference. Each bar represents the mean ± SD of three independent experiments. and versus scrambled RNAi group.

(a)

(b)

(c)