Comprehensive Examination of Hypericum perforatum and Hypericin: Anti-obesity Mechanisms Uncovered by Network Pharmacology and Gene Expression Analysis.

Hypericum perforatum L. or St. John's Wort has long been utilized to treat metabolic and inflammatory conditions in traditional medical systems around the world. Although its antidepressant effects are well known, its role in obesity management remains insufficiently studied. This study aimed to investigate the anti-obesity potential of Hypericum perforatum extract and its major active constituent, hypericin. A combined in vitro and in silico experimental approach was used to explore their effects on adipogenesis, inflammation, insulin signaling and mitochondrial dysfunction. After differentiation, 3T3-L1 preadipocytes were treated with either hypericin (1-8 µM) or Hypericum perforatum extract (25-150 µg/mL). The MTT assay was used to measure cell survival, Oil Red O staining was used to measure lipid accumulation, RT-qPCR was used to measure the expression of 18 obesity-related targets, and ELISA was used to measure protein levels. In silico analyses included target prediction, disease relevance, protein-protein interaction networks, and hub gene identification. GO and KEGG analyses identified key biological pathways. Molecular docking, pharmacokinetics, and toxicity profiling were also performed. Hypericum perforatum extract and hypericin modulated adipogenesis, reduced lipid accumulation, and regulated genes/proteins involved in insulin sensitivity and inflammation. Docking showed strong binding of hypericin to targets such as PPAR-γ, IRS-1, FABP4, and TNF-α. Bioinformatic results indicated activation of the PPAR signaling. Hypericum perforatum and hypericin exhibit multi-target anti-obesity effects via modulation of metabolic, inflammatory, and mitochondrial pathways, supporting their potential as natural therapeutic agents.