Objective: To explore the potential mechanism and pharmacodynamic substances of Huangqin-Tang decoction in the treatment of bacterial infection. Methods: A target gene set and ac-tive compounds of Huangqin-Tang decoction against bacterial infection were obtained using the Traditional Chinese Medicine Systems Pharmacology database (TCMSP) and GeneCards databases. STRING database was utilized for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The compounds-targets network, compounds-targets-pathways net-work (C-T-P) and proteins-proteins interaction (PPI) network were constructed using Cytoscape 3.8.3 software. Molecular docking was performed to visualize the patterns of interactions between the core compounds and key target. Bacteriostatic experiment in vitro was performed to verify the antibacterial activity of core compounds screened. Result: 43 potential targets and 11 active com-pounds of Huangqin-Tang decoction in treatment of bacterial infection were screened. Network analysis indicated that quercetin, kaempferol, wogonin, and beta-carotene may act on 4 core tar-gets, which were TNF, JUN, IL6 and CASP3, and Toll-like Receptor, NOD-like receptor, NF-kappa B, and RIG-I-like receptor signaling pathway and other pathways played a role in anti-bacterial infec-tion. The molecular docking result showed that the key targets had high binding affinity with four core compounds of Huangqin-Tang decoction. In vitro bacteriostatic experimental verified that quercetin, kaempferol had good anti-inflammatory effect. Conclusion: The network pharmacological strategy integrates molecular docking and bacteriostatic experiment in vitro to reveal the thera-peutic effect and potential mechanism of Huangqin-Tang decoction on bacterial infection, which could provide the way to develop new combination medicines for bacterial infection.