Most ginsenosides (the main bioactive substances in ginseng) have been shown to exert potent anti- inflammatory effects, but their anti-inflammatory mechanisms, especially that of ginsenoside F2 (GF2), are incompletely understood. This study was conducted to explore the anti-inflammatory effect of GF2 in an in vitro model of inflammation using LPS-induced RAW264.7 cells. Cell viability analysis revealed that the safe concentration threshold of GF2 to be 100 μmol/L. GF2 significantly inhibited NO release in a dose-dependent manner (0~100 μmol/L) in LPS-induced RAW264.7 cells. GF2 (50, 100 μmol/L) also inhibited the mRNA expression of iNOS, IL-1β, COX2, and TNF-α. GF2 markedly downregulated the protein expression of iNOS, but not that of COX2. Further, treatment with 100 μmol/L GF2 markedly attenuated LPS-induced cell morphology changes in RAW264.7 cells. A western blot showed that GF2 increased PDK1, AKT, and IκB-α protein expression, and decreased AKT phosphorylationand NF-κB nuclear translocation. Our findings demonstrate that GF2 exerts potent anti-inflammatory effects and suggest that these effects are achieved through the AKT/IκB-α/NF-κB signaling pathway. This study can thus provide theoretical support for the development of natural anti-inflammatory drugs.