Figure 2
Fisetin decreases LPS-induced inflammatory mediator and cytokine levels in RAW 264.7 macrophages. RAW 264.7 macrophages (1 × 105 cells/mL) were treated with fisetin (0–8 µM) 2 h before treatment with 500 ng/mL LPS. (A) Total mRNA was isolated at 6 h after 500 ng/mL LPS treatment, and RT-PCR was performed. GAPDH was used as an internal control. (B) Total proteins were isolated at 24 h and western blotting was performed. β-Actin was used as an internal control. (C) The amount of NO production in the culture medium was determined using the Griess Reagent Assay. (D) The amount of PGE2 was determined at 24 h using an ELISA according to the manufacturer’s instructions. (E) Total mRNA was isolated at 6 h and subjected to RT-PCR for IL-6 and TNF-α. The amount of (F) IL-6 and (G) TNF-α was measured at 24 h by an ELISA. Each value indicates the mean ± SEM from three independent experiments. Significant differences among the groups were determined using the Student’s t-test (###p < 0.001 vs. untreated cells) and one-way ANOVA with Bonferroni correction (***p < 0.001, **p < 0.05, and *p < 0.01 vs. LPS-treated cells).
