Abstract: This studyinvestigated the analgesic mechanism of the ethanolic extract of Dalbergia hancei stem by using ultra-high performanceliquid chromatography-quadrupole time-of-flight tandemmass spectrometry (UPLC-Q-TOF-MS/MS) combined withnetwork pharmacology. The chemical constituents of the ethanolic extract, as well as those in the medicated serum, feces, and urine, were analyzed using UPLC-Q-TOF-MS/MS. Active components and analgesic targets were screened through relevant databases. Subsequently, a “component-target” network and a protein-protein interaction (PPI) network were constructed, followed bygene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses. A total of seventy-seven compounds were identified from the ethanolic extract, with thirty-six of them screened as active components. Among these active components, fifteen were confirmed as prototype constituents in vivo. Network pharmacology analysis yielded 139 intersecting targets, from which PTGS2, TIMP1, SERPINE1, and PPARG were identified as the core targets. Pathway enrichment analysis revealed that the analgesic effect was associated with five key pathways, primarily involving the calcium signaling pathway and the serotonergic synapse. The results suggest that Dalbergia hancei stem likely exerts its analgesic effect through a multi-component, multi-target, and multi-pathway mechanism. Active components such as geranylacetone and butein act on multiple targets including PTGS2, and regulate signaling networks involving pathways such as the calcium signaling pathway.

Key words: Dalbergia hancei stem, UPLC-Q-TOF-MS/MS, analgesic, mechanism of action, network pharmacology