Systematically characterized mechanism of Yanhusuo powder ingredient absorbed in rat plasma for treatment osteoarthritis via UPLC-Q-TOF/MS with UPLC-MS/MS and network pharmacology

Background: Yanhusuo powder, also known as Xuanhusuo powder, is a long-standing Chinese herbal formula mainly used in the treatment of ostcoarthritis. Although the clinical effectiveness of Yanhusuo powder has long been acknowledged, its mechanism of action and bioactive components remain unknown. Methods: A novel analytical method combining the use of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and ultra-performance liquid chromatography-triple quadrupole mass spectrometry was applied to profile the formula and absorbed prototype components in plasma after oral administration of Yanhusuo powder. Then, the absorbed constituents were subjected to network pharmacology to predict targets and pathways. AutoDock software was then used for molecular docking studies to screen for potential pharmacodynamic substances. Results: A total of 34 in vitro formula components and 20 in vivo prototype compounds from the various relevant species were successfully separated and identified for the first time. compound-target-pathway analysis revealed that 20 absorbed constituents, 42 target genes and 42 pathways are probably related to the efficacy of Yanhusuo powder against osteoarthritis. The efficacy of Yanhusuo powder mainly involves AKT1, fibronectin 1 and matrix metalloproteinase 9 targets and apoptosis, as well as PI3K-AKT and mitogen-activated protein kinases signaling pathways. According to the results of the molecular docking studies, it can be preliminarily judged that protopine, dehydrocorybulbine and angelicin may be the pharmacologically active substances of Yanhusuo powder. Conclusion: The results provide a scientific basis for understanding the bioactive compounds and the pharmacological mechanism of Yanhusuo powder.