Limonin is a natural triterpenoid derived from the cortex of Phellodendron amurense Ruprecht. Limonin has been reported to have various pharmacological effects such as anti-inflammatory, anti-cancer, anti-bacterial, anti-virus, and anti-oxidative effects. However, the effects and cellular mechanisms of limonin on mast cell-mediated allergic inflammatory responses have remained unclear yet. Therefore, we investigated the effects of limonin on allergic inflammation using phorbol 12-myristate 13-acetate and A23187 (PMACI)-stimulated human mast cell line HMC-1 cells. Limonin inhibited mRNA expression and production of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α) in PMACI-stimulated HMC-1 cells, whereas it did not show cytotoxicity in HMC-1 cells. Limonin also suppressed mRNA expression of Th2-type cytokines (IL-4, IL-5, and IL-13) and chemokines (MCP-1, MIP-1α, MIP-1β, and RANTES) in PMACI-stimulated HMC-1 cells. In addition, limonin inhibited the activation of MAPKs (p38 and JNK) and NF-κB signaling pathways in PMACI-stimulated HMC-1 cells. Moreover, limonin ameliorated the passive cutaneous anaphylaxis reaction of ICR mice. In summary, this study demonstrated that limonin inhibits both allergic inflammatory mediators in HMC-1 cells by suppressing the MAPKs/NF-κB signaling pathway and anaphylaxis reaction in the murine model. Thus, based on these results, limonin might have the potential to become a novel therapeutic agent for allergic inflammatory diseases.