In Vitro Evidence for the Dual Antioxidant and Anti-Inflammatory Roles of Hypericum triquetrifolium in Cancer Therapy: Selective Cytotoxicity Against Pancreatic Cancer Cells

Hypericum species are known for their phytochemical richness and diverse pharmacological activities; however, the biological potential and underlying mechanisms of Hypericum triquetrifolium remain unclear. This study aimed to investigate the phytochemical profile and bioactivities of Hypericum triquetrifolium hydroalcoholic extract (HTE). Plant samples collected from Gelibolu (Çanakkale, Türkiye) were analyzed by HPTLC, revealing the presence of rutin, hyperoside, chlorogenic acid, neochlorogenic acid, and hypericin. Antioxidant activity was evaluated using DPPH, CUPRAC, and FRAP assays, demonstrating strong activity across all tests. Anti-inflammatory effects were assessed in LPS-stimulated RAW264.7 macrophages. HTE significantly reduced nitrite, PGE2, IL-6, ROS, and MDA levels, with the most pronounced effect observed at 0.5 mg/mL (p < 0.05). Cytotoxic and anticancer activities were evaluated in MIA PaCa-2 pancreatic cancer cells and healthy human dermal fibroblasts. The extract exhibited selective cytotoxicity toward cancer cells (IC50 = 0.16 mg/mL), with approximately fivefold lower toxicity in healthy cells. Cell cycle analysis demonstrated G0/G1 arrest, while the Annexin V assay revealed dose-dependent apoptosis (p < 0.0001). DNA damage analysis further supported its anticancer potential. Overall, HTE exhibits potent antioxidant, anti-inflammatory, and selective anticancer activities by reducing oxidative stress, inducing cell cycle arrest, and promoting apoptosis, highlighting its potential as a therapeutic agent.