Hedyotis diffusa Willd.-Scutellaria barbata D.Don herb pair induces ferroptosis in gastric cancer by inhibiting the PI3K/AKT/HIF-1α pathway.

The herb pair comprising Hedyotis diffusa Willd. and Scutellaria barbata D.Don (HD-SB) is widely utilized in oncological therapies. Nevertheless, the mechanisms underlying its suppressive effects on gastric cancer (GC) remain inadequately elucidated. This investigation aims to delineate the therapeutic efficacy and molecular targets of HD-SB against GC through an integrative approach combining high-performance liquid chromatography-mass spectrometry (HPLC-MS), network pharmacology, molecular docking, and experimental validation. Thirty-three bioactive compounds were retrieved for HD-SB, yielding 288 putative therapeutic targets relevant to GC. Protein-protein interaction (PPI) network analysis highlighted 23 pivotal targets, including AKT1 and HIF-1α. Functional enrichment analyses via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways indicated significant involvement of the PI3K/AKT signaling cascade and transcription factor activity. Molecular docking studies demonstrated strong binding affinities between principal HD-SB constituents and core targets AKT1 and HIF-1α. Bioinformatic analysis revealed elevated mRNA and protein expression levels of AKT1 and HIF-1α in GC tissues relative to normal gastric tissues, with increased AKT1 expression correlating with poorer patient prognosis. In vitro assays confirmed that HD-SB markedly inhibited GC cell proliferation, while in vivo experiments substantiated its antitumor efficacy. Mechanistically, HD-SB was shown to induce ferroptosis in GC cells by suppressing the PI3K/AKT/HIF-1α pathway. Collectively, these findings suggest that HD-SB exerts antineoplastic effects in GC by promoting ferroptosis through modulation of the PI3K/AKT/HIF-1α signaling axis, underscoring its potential clinical utility in GC management.