Background: Mesuaferrin-A, bio active flavonoid has been isolated from bark of Mesua ferrea L. shown to inhibit NO and TNF –α production in LPS induced RAW 264.7 cells. These flavonoids possess biological properties, such as antioxidant, anti-inflammatory, antimicrobial and anticancer activities. Objective: The main aim of the study is to investigate the mechanisms underlying the anti-inflammatory effects of Mesuaferrin-A on lipopolysaccharide- (LPS) induced inflammation in RAW 264.7 cells. Methods: To evaluate the anti-inflammatory effect of Mesuaferrin-A, on lipopolysaccharide (LPS)-stimulated inflammation in mouse RAW264.7 cells. The inflammatory mediator Cyclooxygenase-2 (COX-2) and regulatory transducing factor like nuclear factor-κB (NF-κB), p38 MAPK (p38) and AKT were analysed by Western blotting. Results: The results showed that Mesuaferrin A (50 μg/mL) significantly inhibited the COX-2 gene and protein expressions through prevent the nuclear translocation of NF-κB by inhibiting the phosphorylation of IkBα and also inhibited activation of the p38 MAPK phosphorylation and Akt in LPS-treated RAW 264.7 cells. Conclusion: Mesuaferrin A, inhibited COX-2 expression through the down regulation of NF-κB-dependent transcriptional factor suggesting that Mesuaferrin A bioactive flavonoids is the important constituent in the bark of Mesua ferrea L and has a potential novel anti-inflammatory agent targeting on the NF-κB signaling pathway, to the treatment of inflammatory diseases.