Exploration of the effect and mechanism of Eclipta prostrata L. against gastric cancer based on network pharmacology and experimental verification

The present study aimed to explore the therapeutic effects and underlying mechanisms of the active constituents of Eclipta prostrata L. in gastric cancer (GC) and to identify potential targets for its prevention and treatment. Active compounds and their corresponding targets were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform. In comparison, GC‑related targets were retrieved from GeneCards and Online Mendelian Inheritance in Man databases. Common targets were identified using Venny 2.1.0. Cytoscape 3.10.2 was then employed to construct a traditional Chinese medicine component‑target‑disease network and a protein‑protein interaction network. Core compounds and targets were further assessed through molecular docking, and the potential mechanisms were investigated using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. The biological activity of luteolin, the principal active component of Eclipta prostrata L., was experimentally validated in AGS cells using Cell Counting Kit‑8 assays, wound‑healing (scratch) assays, Transwell migration assays, flow cytometry and western blotting. A total of 10 active ingredients and 166 shared targets were identified. Among them, quercetin, luteolin and acacetin were identified as key bioactive constituents, whereas AKT serine/threonine kinase 1 (AKT1), TNF and hypoxia‑inducible factor‑1 (HIF‑1A) were recognized as central targets. Molecular docking analysis revealed strong binding affinities between the core compounds and these targets. Enrichment analyses further indicated that the therapeutic effects of Eclipta prostrata L. may involve pathways related to oxidative stress, the phosphatidylinositol 3‑kinase‑protein kinase B signaling pathway, and the HIF‑1 signaling pathway. Among the identified compounds, luteolin showed the most pronounced inhibitory effects on GC cells and was therefore selected for further mechanistic investigation. In vitro experiments demonstrated that luteolin significantly suppressed GC cell proliferation and migration, reduced AKT phosphorylation and HIF‑1A expression, and did not induce apoptosis. These findings suggest that luteolin may exert its anti‑GC effects primarily by modulating the AKT1/HIF‑1A signaling axis. The present study provides experimental support for the therapeutic potential of Eclipta prostrata L.‑derived compounds and offers insights into novel molecular targets for GC treatment.