Exploring the potential mechanisms of action of Gentiana veitchiorum Hemsl. extract in the treatment of cholestasis using UPLC-MS/MS, systematic network pharmacology, and molecular docking

Abstract Ethnopharmacological relevance Gentiana veitchiorum Hemsl. (GV) has a long history in Tibetan medicine for the treatment of the hepatobiliary disease, cholestasis. However, the mechanisms mediating its efficacy in the treatment of cholestasis have not been determined. Purpose To elucidate the mechanisms of action of GV in the treatment of cholestasis, an integrated approach combining ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis with network pharmacology and molecular docking was established. Materials and methods A comprehensive analysis of the chemical composition of GV was achieved by UPLC-MS/MS. Subsequently, a network pharmacology method that integrated target prediction, a protein-protein interaction (PPI) network, gene set enrichment analysis and component-target-pathway network was established. The network pharmacological results were validated through molecular docking analysis. Results The UPLC-MS/MS analysis identified twenty compounds in GV. Network pharmacology identified 299 targets for the components of GV and 2999 targets for cholestatic diseases, from which 103 intersectional targets were obtained by Venn diagram. A PPI network was constructed to screen the following core proteins: AKT1, GAPDH, TNF, IL6, VEGFA, and CASP3. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses results suggested that GV may treat cholestatic disease through biological pathways related to proteolysis, cytosol, caspase complex, enzyme binding, and pathways in cancer, lipid and atherosclerosis, HIF-1, TNF, IL-17 and other key signaling pathways. The results were validated by molecular docking studies, which indicated good binding affinities between core compounds and targets, with similar binding energies to those of the positive control drugs. Conclusions In this study, UPLC-MS/MS analysis and network pharmacology were used in conjunction to provide potential mechanisms of action of GV in the treatment of cholestasis.