ETHNOPHARMACOLOGICAL RELEVANCE: Achillea alpina L. is a traditional herbal medicine with a long history, which is often used to detoxify and relieve pain. Achillea alpina L. essential oil (AHO) is extracted from the aboveground part of the Achillea alpina L. The role of AHO on the in vivo anti-inflammatory effects remains unclear. AIM OF THE STUDY: To explore the anti-inflammatory effect and interaction mechanism of AHO in zebrafish tail fin model. MATERIALS AND METHODS: The chemical components of AHO were first identified utilizing gas chromatography-mass spectrometry (GC-MS). A zebrafish tail fin model was employed to evaluate the anti-inflammatory effect of AHO by observing the numbers of neutrophils and the expression levels of pro-inflammatory cytokines. The combined application of transcriptomics and metabolomics helped us to explore the potential anti-inflammatory mechanism of AHO, and the expression of core gene was verified by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The principal constituents of the AHO included bicyclo sesquiphellandrene (11.99%), α-thujene (6.19%), 1-methyl-7-isopropyl naphthalene (5.90%), and β-elemene (5.58%). AHO exhibited potent anti-inflammatory properties by dramatically inhibiting the migration of neutrophils to the tail fin amputation site, along with autophagy linked to inflammation. Moreover, AHO had an excellent regulatory influence on the expression of pro-inflammatory cytokines, including tumor necrosis factor alpha, interleukin 6, and interleukin 1β. Furthermore, transcriptome and metabolomic analyses identified a crucial gene and fourteen significant metabolites influenced by AHO in relation to inflammation. The investigation demonstrated that AHO modulated the inflammatory response via influencing amino acid and glucose metabolism. CONCLUSION: In this study, AHO has excellent anti-inflammatory effects and shown remarkable regulatory effects on the expression of immune cells and pro-inflammatory factors in vivo, which is highlighting the necessity for more research and development as a potential anti-inflammatory drug.