Background and Objective: Refractory ulcerative colitis has few effective drugs and an unknown etiology. Artesunate, an artemisinin derivative, may decrease colon inflammation in animal models via immunomodulation, although its targets and mechanisms are unclear. This study predicted artesunate targets, processes and effectiveness in treating ulcerative colitis using network pharmacology, molecular docking and animal trials. Materials and Methods: Artesunate and ulcerative colitis gene targets were searched in databases. The KEGG pathway enrichment and gene ontology analysis evaluated possible targets. Artesunate was evaluated in C57 mice with 3% dextran sodium sulphate-induced ulcerative colitis. Target gene expression was shown by western blotting, Haematoxylin and Eosin (H&E) staining, RT-PCR and immunohistochemistry. Results: Overall, network pharmacology evaluated 154 artesunate drug targets and 2,104 ulcerative colitis targets. The intersection of these objectives has 29 candidates. These targets controlled apoptosis, p53 and JAK/STAT signalling. The final validation targets were IL-6 and TP53 using molecular docking studies. In mouse models, artesunate reduced dextran sodium sulphate-induced weight loss, colon length, disease activity index and pathological scores and increased TP53 and decreased interleukin-6 expression in ulcerative colitis colon tissues. In the colon, artesunate activated Bax and Caspase-3 and downregulated STAT3 and Bcl-2. Conclusion: Network pharmacology and molecular docking identified artesunate’s ulcerative colitis targets and processes. Artesunate decreased colon edema and damage in 3% dextran sodium sulphate-treated rats. To induce these effects, TP53 and interleukin-6 may control apoptosis and inflammation. Current findings might lead to ulcerative colitis medications.