Integrative network pharmacology, machine learning, and experimental validation reveal the protective potential and potential molecular mechanisms of orientin in DSS-induced colitis

Background Ulcerative colitis (UC) is a chronic and recurring inflammatory disease affecting the colonic epithelium. Orientin, a natural flavonoid C-glycoside, shows various anti-inflammatory and antioxidant properties. However, the effects of orientin on UC and the related molecular mechanisms have not been thoroughly studied. Methods UC-related genes were identified from GeneCards, OMIM, and TTD and analyzed using the GSE87466 dataset, followed by GO and KEGG enrichment analyses. Four machine-learning algorithms were applied to identify candidate targets. Molecular docking and 100-ns molecular dynamics simulations followed by MM-PBSA binding free-energy analysis were performed for the identified target–Orientin complexes. ROC was used to evaluate model performance. In vivo, DSS-induced colitis mice were treated orally with Orientin (10 mg/kg/day), and disease severity, histopathological changes, target gene expression, and pro-inflammatory cytokines were evaluated. Results Four central targets, such as ABCB1, PPARGC1A, FAP and LYN, were selected by all the machine learning techniques. The AUC values of the four-gene model in the training set and the validation set were all greater than 0.8. In vivo experiments show that Orientin can considerably attenuate DSS-induced weight loss, reduce the Disease Activity Index (DAI), reverse DSS-induced colon shortening and decrease the histopathological damage caused by DSS. Additionally, Orientin can restore the abnormal mRNA levels of ABCB1, PPARGC1A, FAP and LYN. It can also decrease the high mRNA expression levels of IL - 6, IL - 1β and TNF-α in the colonic tissues, which are induced by DSS. Conclusion The results of this study show that Orientin has protective effects on colitis caused by DSS via a multi-target regulatory process. ABCB1 and PPARGC1A may represent important candidate targets of Orientin.