Integrated Network Pharmacology, Metabolomics, And in vivo Validation of Liangxue Dihuang Decoction In DSS-Induced Ulcerative Colitis Mice.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by persistent intestinal inflammation and metabolic dysregulation. Traditional Chinese medicine (TCM) formulas are widely used in clinical practice for inflammatory disorders; however, systematic experimental approaches to elucidate their multi-component and multi-target mechanisms remain limited. This article presents a reproducible experimental protocol integrating network pharmacology, untargeted metabolomics, and in vivo validation to investigate the anti-inflammatory effects of Liangxue Dihuang Decoction (LXDHD) in a dextran sulfate sodium (DSS)-induced mouse model of UC. The workflow includes preparation of the herbal decoction, DSS-induced colitis modeling, disease activity assessment, histopathological and hematological evaluation, serum cytokine measurement, LC-MS-based metabolomic profiling, and computational network analysis. Using this integrated approach, DSS-induced colitis was successfully established, and LXDHD treatment resulted in measurable improvements in the disease activity index, colon length, histopathological damage, inflammatory cytokine levels, and metabolic profiles. Network pharmacology and metabolomics analyses further demonstrated that this protocol enables the identification of key targets, pathways, and metabolites associated with therapeutic intervention. This visualized and standardized workflow provides a practical methodological framework for studying the pharmacological mechanisms of multi-component TCM formulas in inflammatory bowel disease and can be adapted to other herbal prescriptions and disease models.