Integrating Network Pharmacology and Metabolomics to Elucidate the Mechanism of Cryptotanshinone Against Platelet...

Cryptotanshinone (CTS), an antiplatelet compound from Salvia miltiorrhiza, exhibits in vitro potency comparable to aspirin. This study integrated network pharmacology and metabolomics to elucidate its underlying mechanisms. An acute blood stasis model was induced in Sprague-Dawley rats using epinephrine and ice-water immersion. Animals were assigned to seven groups. Platelet aggregation was measured turbidimetrically using arachidonic acid (AA) and adenosine diphosphate (ADP) as agonists. Core targets were predicted by network pharmacology, differential metabolites were screened, and pathways were enriched using untargeted metabolomics. Integrated analysis identified shared pathways and key targets, validated by molecular docking. AA- and ADP-induced aggregation was significantly increased in model rats versus the blank group. CTS at all doses markedly inhibited aggregation in a dose-dependent manner. Network pharmacology identified 15 core targets. Metabolomics identified 51 differential metabolites enriched in seven pathways, including glycerophospholipid and butanoate metabolism. Integrated analysis revealed five common pathways: linoleic acid metabolism, arginine biosynthesis, AA metabolism, glutathione metabolism, and drug metabolism-and four key targets (CYP3A4, NOS3, PTGS2, and GSTP1). Molecular docking showed strong binding energies (<-9 kcal/mol) between CTS and these targets. CTS inhibits platelet aggregation by regulating CYP3A4, NOS3, PTGS2, and GSTP1 and intervening in five metabolic pathways, supporting its potential as an anti-platelet agent.