The aim of this study was to investigate the effect of glycyrrhizin on LPS-induced endotoxemia in mice and clarify the possible mechanism.An LPS-induced endotoxemia mouse model was used to confirm the anti-inflammatory activity of glycyrrhizin in vivo.In vitro, RAW264.7 cells were stimulated with LPS in the presence or absence of glycyrrhizin.The expression of cytokines was determined by ELISA.Toll-like receptor 4 (TLR4) was determined by Western blot analysis.Nuclear factor-kB (NF-κB) and Interferon regulatory factor 3 (IRF3) activation were detected by Western blotting and luciferase assay.Lipid raft staining was detected by immunocy to chemistry.In vivo, the results showed that glycyrrhizin can improve survival during lethal endotoxemia.In vitro,glycyrrhizin dose-dependently inhibited the expression of TNF-α, IL-6, IL-1β and RANTES in LPS-stimulated RAW264.7 cells.Western blot analysis showed that glycyrrhizin suppressed LPS-induced NF-κB and IRF3 activation.However, glycyrrhizin did not inhibit NF-κB and IRF3 activation induced by MyD88-dependent (MyD88 ,IKKβ) or TRIF-dependent (TRIF, TBK1) downstream signaling components.Moreover, glycyrrhizin did not affect the expression of TLR4 and CD14 induced by LPS.Significantly, we found that glycyrrhizin decreased the levels of cholesterol of lipid rafts and inhibited translocation of TLR4 to lipid rafts.Moreover, glycyrrhizin activated ABCA1, which could induce cholesterol efflux from lipid rafts.Glycyrrhizin exerts an anti-inflammatory property by disrupting lipid rafts and inhibiting translocation of TLR4 to lipid rafts, thereby attenuating LPS-mediated inflammatory response.Learning the anti-inflammatory mechanism of glycyrrhizin is crucial for the anti-inflammatory drug development.