Mechanism of Xianglian pills in treatment of ulcerative colitis based on network pharmacology and experimental verification

AimThis study aims to predict the potential targets and related signaling pathways of Xianglian pills against ulcerative colitis (UC) using UHPLC-TOF-MS, network pharmacology and molecular docking, and to verify the mechanism of action through animal experiments.MethodsThe intestinal components of Xianglian pills were analyzed by UHPLC-TOFMS, and the targets and related signaling pathways of UC were predicted by network pharmacology and verified by molecular docking. The UC model mice were subsequently established, and different doses of Xianglian pills were administered to investigate the changes of colon tissue phenotype and disease activity index. HE staining was used to observe pathological changes in colon tissue, and inflammatory factors in serum were detected by ELISA. The key targets in colon tissue were detected by Western blot and immunofluorescence.ResultsA total of 25 intestinal components of Xianglian pills were identified, which were predominantly alkaloids. Network pharmacological results showed that 495 compound-disease targets were found, and 95 core targets were screened out by PPI. KEGG enrichment analysis showed that Xianglian pills may play a role in the treatment of UC through JAK2-STAT3, Th17 cell differentiation, HIF-1, IL-17, and other signaling pathways. Molecular docking showed that the key active ingredients of Xianglian pills had a stable binding conformation with the core proteins. The results of animal experiments showed that Xianglian pills could significantly improve the colon pathological injury of UC model mice. ELISA indicated that Xianglian pills could significantly reduce the serum levels of IL-17 and IL-23 and increase the serum levels of TGF-β and IL-10 in UC model mice. Western blot and immunofluorescence showed that Xianglian pills could significantly inhibit the protein expression levels of p-JAK2/JAK2, p-STAT3/STAT3 and RORγt in colon tissue of UC model mice, and increase the protein expression level of Foxp3.ConclusionXianglian pills may play a role in the treatment of UC by inhibiting JAK2-STAT3 pathway, thereby improving Th17/Treg immune imbalance.