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Our previous study showed that the extract of Inula japonica Thunb (I.japonica) has anti-inflammatoryandanti-asthmaticactivities.Recently,weisolated1,6-dihydroxy-4 H-1,10-secoeudesma-5(10),11 (13)-dien-12,8-olide (SE),6-isobutyryloxy-1-hydroxy-4 H-1,10-secoeudesma-5(10),11(13)-dien-12,8-olide(IBSE),and6-isovaleryloxy-1-hydroxy-4 H-1,10-secoeudesma-5(10),11(13)-dien-12,8-olide (IVSE) from I.japonica, and investigated their inhibitory effects on nitric oxide (NO) production in LPS-stimulated RAW264.7 cells.Among them, IVSE displayed the most potent inhibition of NO production.Then we investigated the potential anti-inflammatory mechanism.IVSE dramatically decreased the expression of inducible nitric oxide synthase (iNOS), reduced the translocation of the Nuclear factor B (NF-B)subunit p65 into the nucleus by interrupting the phosphorylation and degradation of inhibitor kappa B(IB)-, and inhibited the activation of the upstream mediator IkB kinase(IKK).Furthermore, our results indicated that IVSE inhibited the phoshorylation of mitogen-activated protein kinases (MAPKs) including extracellular regulated kinases (ERK1/2), c-Jun N-terminal kinases (JNK) and p38.Taken together, our results showed that IVSE exhibited anti-inflammatory activity by inhibiting NO production, in which inactivation of NF-B and MAPKs might be involved.