Abstract: Through network pharmacology and molecular docking technology, this study investigated the active components and mechanism of mulberry leaves, a food-medicine homologous plant, in treatment of hyperuricemia. The primary components, targets and pathways underwent systematic evaluation, moreover in vitro experiments were constructed to further analyze. The results demonstrated that a total of 16 active components associated with hyperuricemia, including Oxysanguinarine, moricin D and Inophyllum E were identified through the screening of 269 active components derived from mulberry leaves. Subsequently, 80 targets related to hyperuricemia were identified. Furthermore, 15 core targets were screened out, respectively glyceraldehyde 3- phosphate dehydrogenase (GAPDH), tumor necrosis factor (TNF), estrogen receptor 1 (ESR1), etc. A total of 321 biological process entries, 46 cell components entries and 108 molecular function entries were received by conducting Gene ontology (GO) analysis. 97 signal pathways related to hyperuricemia were obtained through Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, among which metabolic pathways and nucleic acid metabolic pathways played an anti-hyperuricemia role. Molecular docking indicated that the binding energies of the key components of mulberry leaves to the top 6 core targets were less than or equal to -6 kcal/mol, and the binding energies of Oxysanguinarine to the three core target proteins ESR1, CASP3 and PTGS2 were found to be the highest. Further research on the mechanism of Oxysanguinarine in treating hyperuricemia in vitro presents that this substance can reduce the activity of xanthine oxidase, and enhance the uric acid excretion ability of HK-2 and HEK-293T cells by regulating the proteins such as urate transporter 1 (URAT1) and adenosine triphosphate binding cassette transporter G2 (ABCG2). In summary, present investigation reveals that mulberry leaves are efficacious in the treatment of hyperuricemia through a multi-targets and multi-channels, which provides scientific basis for animal experiments and development of uric acid-reducing foods from mulberry leaves.