AIM: To explore the core target and mechanism of α-Linolenic acid (ALA) in improving neuroinflammation through network pharmacology combined with in vitro experiments.METHODS: Pharmacological studies have shown that ALA has anti-inflammatory, antioxidant, and neuroprotective properties. The targets of α-Linolenic acid were obtained from PharmMapper and Swiss Target Prediction databases, the targets of neuroinflammation were searched from GeneCards, TTD and OMIM databases, and the potential targets of ALA and neuroinflammation were obtained from Wayne diagram.Protein interaction network (protein - protein interaction, PPI) of potential targets was constructed by STRING website, and the core targets in PPI were screened by Cytoscape 3.8.0 software.At the same time, potential targets are imported into DAVID database, GO and KEGG data were obtained and the results were visualized.Autodock vina and Pymol software were used to dock the selected core targets with ALA and visualize the results. An in vitro model of neuroinflammation was constructed, and cell growth status, oxidative stress, and migration or repairing capacity were determined by CCK-8 analysis, SOD, MDA and cell scratches, and the expression of IL-6, iba 1, COX-2 (PTGS2), and iNOS proteins was determined by ELISA or Western blot experiments.RESULTS: Network pharmacology analysis revealed 46 potential targets of ALA for neuroinflammation, and 10 core targets, including IL-6 and PTGS 2. With 232 entries enriched by GO enrichment analysis and 70 signaling pathways enriched by KEGG enrichment analysis, molecular docking showed that ALA can form hydrogen bonding with COX-2.Experiments showed that ALA could improve cell viability, alleviate cell oxidative stress levels, and promote cell migration and motor repair in an in vitro model of neuroinflammation. CONCLUSIONS: ALA may improve neuroinflammation by alleviating oxidative stress and inhibiting IL-6 and COX-2 protein expression.