This study explored the effect and mechanism of Dracocephalum moldavica L. on acute kidney injury (AKI) based on network pharmacology, molecular docking and animal experiments. Through online pharmacological studies, the effective components and targets of Dracocephalum moldavica L. in the treatment of AKI were obtained. Seventeen active components and 60 intersection targets were screened out, among which there were 12 core targets. Through the analysis of the function of Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, it was screened out that it was mainly related to pathways such as cancer. Molecular docking was conducted between the active ingredients and the core targets. The results showed a strong association with matrix metalloprotein 9 (MMP9) and glycogen synthase kinase-3β (GSK-3β). The experiment selected the extract of Dracocephalum moldavica L. as the therapeutic drug and randomly divided ICR mice into normal group, model group, low, medium, and high (90, 180, 360 mg·kg^-1) dose groups of Dracocephalum moldavica L. extract, amifostine group (200 mg·kg^-1), and Uremic Clear Granules group (3 750 mg·kg^-1). Intragastric administration was given 7 days in advance. On the 7^th day, the amifostine group was injected with amifostine solution. Two hours after the end of intragastric administration on the 7^th day, except for the normal group, the other groups were intraperitoneally injected with cisplatin (CDDP) solution to establish an AKI mouse model. Samples were collected 72 hours after modeling. This experiment was approved by the Animal Ethics Committee of Xinjiang Institute of Materia Medica (approval number: XJIMM-20240802). The organ indices of liver, spleen, and kidney were calculated; the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) in mouse serum, glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) in mouse kidney were determined by kits; the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in mouse kidney were detected by ELISA kits; the pathological changes of the kidney were observed by hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining; and the expression of MMP9, GSK-3β, β-catenin, as well as Bcl-2 associated X-protein (Bax) and B-cell lymphoma-2 protein (Bcl-2) in renal tissue was verified by Western blot. The experimental results showed that the extract of Dracocephalum moldavica L. could significantly improve the organ indices of spleen, liver, and kidney in AKI mice; significantly reduce the levels of SCr and BUN in blood; reduce the levels of TNF-α, IL-6, and IL-1β; up-regulate the levels of GSH-PX and SOD; improve the pathological damage of kidney; and regulate the expression of MMP9, GSK-3β, β-catenin, Bax and Bcl-2 in renal tissue. It suggests that the extract of Dracocephalum moldavica L. may exert a protective effect on AKI caused by CDDP through regulating the expression of the GSK-3β/β-catenin signaling pathway.