Abstract: OBJECTIVE To explore the mechanism of diuretic effect of Lepisorus ussuriensis (Regel et Maack) Ching and to compare the diuretic effect of Lepisorus ussuriensis (Regel et Maack) Ching and its regenerated plants. METHODS Using network pharmacology to explore the diuretic mechanism of Lepisorus ussuriensis (Regel et Maack) Ching; AutoDock Vina1.1.2 was applied to acquire the molecular docking result of active components and core targets, then Pymol2.5.0 software was used to visualize the optimal results; the urine volume of mice was measured within 4 h after oral administration by in vivo experiments; to evaluate the diuretic effects of the Lepisorus ussuriensis (Regel et Maack) Ching and its regenerated plants. RESULTS A total of 1 791 drug targets were obtained from 41 candidate compounds of Lepisorus ussuriensis (Regel et Maack) Ching, and to map the 327 drug targets obtained after deleting repeated targets and the 1 324 disease targets retrieved from the GeneCards database with "Diuretic" as the key word. A total of 119 intersecting genes were obtained. The top 6 core proteins of PPI network nodes were MAPK3, VEGFA, AKT 1, IL6, ALB and TNF. The top three candidate compounds were baicalein(44), diosmetin(43) and kaempferol(43). The results of GO function analysis and KEGG pathway enrichment analysis showed that the diuretic effect of Lepisorus ussuriensis (Regel et Maack) Ching was mainly related to response to toxic substance, membrane raft et al, the main signal pathways involved were neuroactive ligand-receptor interaction, HIF-1 signaling pathway, nitrogen metabolism and so on. The results of molecular docking revealed that baicalein, diosmetinand, kaempferol had better spontaneous binding effect with the key target. The result of in vivo experiments showed that Lepisorus ussuriensis (Regel et Maack) Ching(9.5 g·kg^-1) increased the urinary volume(1.26-fold), as well as the excretion of K⁺(1.31-fold) and Cl^-(1.24-fold). The regenerated plants(9.5 g·kg^-1) increased the urinary volume(1.26-fold), as well as the excretion of K⁺(1.29-fold) and Cl^-(1.26-fold). CONCLUSION The mechanism of diuretic action of Lepisorus ussuriensis (Regel et Maack) Ching is studied by network pharmacology, and the characteristics of multi-component, multi-target and multi-pathway were verified. Lepisorus ussuriensis (Regel et Maack) Ching and its regenerated plants show the same diuretic activity through in vivo experiments.