Objective To explore the therapeutic effect and mechanism of Biyu Shibai xiji (BYSB) on eczema. Methods Using network pharmacology methods, we collected compound-related targets and eczema-related gene targets, identified the intersection targets between drugs and diseases, constructed protein-protein interaction networks as well as drug-compound-target-disease networks, and performed KEGG pathway enrichment analysis to identify pathways associated with eczema treatment. An eczema model was established by repeatedly stimulating the skin of male KM mice with 2,4-dinitrochlorobenzene (DNCB). A total of 48 male KM mice were randomly divided into control group (Control), model group (Model), positive drug group (Positive), high dose group (BYSB-H, 2g·kg-1), middle dose group (BYSB-M, 1g·kg-1), low dose group (BYSB-L, 0.5g·kg-1). Except for the blank group, the mice in the other groups were repeatedly stimulated with 2,4-dinitrochlorobenzene to establish eczema models. Each group was given corresponding test drugs for 14 days, and samples were taken. The scores of skin lesions on the back of mice were observed and recorded. Hematoxylin-eosin (H&E)staining was used to observe the pathological changes of skin tissue on the back of mice. The mast cells in the skin tissue of the mouse''s back were observed by toluidine blue (TB) staining.The levels of interleukin-4 (IL-4), interferon-γ(IFN-γ) and immunoglobulin E (IgE) in serum of mice were detected by enzyme-linked immunosorbent assay (ELISA). The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) in skin tissue homogenate of mice were determined by biochemical kit. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in skin homogenate of mice were determined by ELISA. The expression of caspase-3 protein in skin lesions was determined by immunohistochemistry (IHC). The expression of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and nuclear factor-κB (NF-κB) protein in skin lesions was detected by Western blot (WB). Results Network pharmacology analysis identified 130 potential targets for the treatment of eczema by the key herbs Portulaca oleracea and Sophora flavescens, with IL6, CASP3, NFKBIA, TNF, and RELA selected as core targets associated with eczema. KEGG pathway enrichment analysis revealed several pathways related to eczema treatment, including the PI3K-Akt signaling pathway, HIF-1 signaling pathway, NF-κB signaling pathway, and chemokine signaling pathway. Animal experiment results showed that all doses of BYSB improved inflammatory infiltration in the skin (P<0.01, P<0.05) and reduced the number of degranulated mast cells (P<0.01). Compared with the model group, Biyu Shibai Lotion could reduce the expression levels of IL-4, IFN-γ and IgE in serum (P<0.01), and callback the ratio of IFN-γ/IL-4 (P<0.01). The levels of SOD and GSH-PX in the homogenate of skin lesions were increased, and the levels of IL-6 and TNF-αwere decreased. Reduce the expression level of Caspase-3 protein. The expression of TLR4, MyD88 and NF-κB protein in skin lesions was significantly reduced (P<0.01).Conclusion Biyu Shibai Lotion has a good therapeutic effect on DNCB-induced eczema model in mice. Its mechanism may be to regulate immune balance, inhibit TLR4/MyD88/NF-κB inflammatory pathway and enhance antioxidant capacity, so as to achieve the purpose of treating eczema.