Mangostenone F (MF) is a natural xanthone isolated from Garcinia mangostana. However, little is known about the biological activities of MF. This study was designed to investigate the anti-inflammatory effect and underlying molecular mechanisms of MF in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. MF dose-dependently inhibited the production of NO, iNOS, and pro-inflammatory cytokines (TNF-${alpha}$, IL-6, and IL-$1{eta}$) in LPS-stimulated RAW264.7 macrophages. Moreover, MF decreased the NF-${kappa}B$ luciferase activity and NF-${kappa}B$ DNA binding capacity in LPS-stimulated RAW264.7 macrophages. Furthermore, MF suppressed the NF-${kappa}B$ activation by inhibiting the degradation of $I{kappa}B{alpha}$ and nuclear translocation of p65 subunit of NF-${kappa}B$. In addition, MF attenuated the AP-1 luciferase activity and phosphorylation of ERK, JNK, and p38 MAP kinases. Taken together, these results suggest that the anti-inflammatory effect of MF is associated with the suppression of NO production and iNOS expression through the down-regulation of NF-${kappa}B$ activation and MAPK signaling pathway in LPS-stimulated RAW264.7 macrophages.