Abstract: Objective This study aimed to explore the therapeutic targets and mechanisms of Broussonetia papyrifera (BP) in the treatment of atopic dermatitis (AD) through network pharmacology, molecular docking, and animal experiments. Methods An AD mouse model was established through 2, 4-dinitrochlorobenzene (DNCB) induction. The mice were divided into five groups: normal control group (Nor), model group (Mod), dexamethasone group (DEX), low-dose BP extract group (L-BP), and high-dose BP extract group (H-BP), and treated for 12 consecutive days. The therapeutic effects of BP on AD were evaluated by observing the skin damage, scoring the dermatitis, and conducting histopathological staining. The active components of BP and their targets were predicted using the HERB, TCMSP, and SwissTargetPrediction databases. Targets related to AD were screened using the GeneCards, DisGeNET, and OMIM databases. The common targets of BP and AD were identified by Venny and visualized using a Venn diagram. Protein-protein interaction (PPI) network analysis was performed using STRING and Cytoscape software, and the top 10 targets with the highest degree values were identified as core targets. The possible mechanism of BP in treating AD was analyzed using Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. The binding affinity between the active components of BP and the core targets was assessed using AutoDock software. Finally, Western blot experiments were conducted to verify the predictions made by network pharmacology. Results The in vivo experimental results showed that, compared with the model group, BP significantly increased the epidermal thickness of DNCB-induced AD mice, reduced the dermatitis score, and improved AD symptoms. Network pharmacology analysis identified 47 common targets from 115 BP targets and 1 815 AD targets. PPI network topology analysis determined 10 core targets, including ESR1, AKT1, PTGS2, HIF1A, and NF-κB1. Molecular docking results indicated that the active components of BP had good binding affinity with these core targets. GO function enrichment analysis revealed 74 molecular functions, 22 cellular components, and 167 biological processes. KEGG pathway enrichment analysis suggested that the PI3K-AKT and the NF-κB signaling pathway might be important pathways for BP in treating AD. Animal experiments demonstrated that BP could inhibit the activation of the PI3K/AKT and NF-κB signaling pathways by reducing the phosphorylation levels of PI3K, IKB, p65, and AKT. Conclusion BP exerts its therapeutic effects on AD by modulating the PI3K/AKT and NF-κB signaling pathways, which provides a scientific basis for further in-depth research and potential clinical application of BP in the treatment of AD.