OBJECTIVE: To investigate the mechanisms of 6-hydroxygenistein (6-OHG) in the treatment of high-altitude hypoxia-induced lung injury. METHODS: The intersection targets of 6-OHG and high-altitude hypoxia-induced lung injury were identified using databases, including Swiss Target Prediction, SuperPred, GeneCards, and OMIM. The STRING database and Cytoscape software were used to construct a protein interaction network for the intersection targets of drugs and diseases, and targets with degree values greater than the median were identified as key targets. GO and KEGG enrichment analyses of key targets were performed using the DAVID database to identify relevant signaling pathways. The Maestro 13.7 software was used for molecular docking validation. A large hypobaric hypoxic chamber was used to establish a high-altitude lung injury model in mice. A total of 42 male BALB/c mice were randomly assigned to 3 groups ( RESULTS: Key targets such as serine/threonine protein kinase 1 (AKT1), HIF-1α, epidermal growth factor receptor (EGFR), matrix metalloproteinase 9 (MMP9), and peroxisome proliferator-activated receptor A (PPARA) were identified. GO and KEGG enrichment analyses showed that the targets of 6-OHG in the treatment of high altitude hypoxia-induced lung injury were mainly involved in PI3K/AKT, HIF-1α/VEGF, tumor necrosis factor (TNF), and other signaling pathways. The results of animal experiments demonstrated that compared with the model group, the 6-OHG group showed significant improvement in the pathological damage of lung tissues induced by high altitude hypoxia, presenting statistically significant differences in the levels of MDA, H CONCLUSION: 6-OHG may alleviate lung injury induced by high altitude hypoxia in mice by activating the PI3K/AKT signaling pathway and inhibiting the HIF/VEGF signaling pathway.