Mechanism of cardamonin in inhibiting pulmonary fibroblast senescence through the mTOR-ROS axis

Cardamonin, a natural flavonoid compound, exhibits potential anti-aging properties, yet its precise mechanisms still remain unclear. In this study, we established cellular senescence models using bleomycin and H₂O₂ and treated the cells with varying concentrations of cardamonin to investigate its anti-senescence effects and underlying mechanisms. Senescence-associated β-galactosidase (SA-β-gal) staining was employed to assess senescent phenotypes, while immunofluorescence was used to detect DNA damage levels. Intracellular reactive oxygen species (ROS) levels were measured using the DCFH-DA probe, and Western blot was performed to analyze the expression of p53, p21, collagen, α-smooth muscle actin (α-SMA), mechanistic target of rapamycin (mTOR), and p-mTOR. To further validate the mechanistic target, MHY1485 was utilized to activate the mTOR pathway and evaluate its regulatory impact on senescence phenotypes. The results demonstrated that cardamonin significantly alleviated bleomycin- and H₂O₂-induced cellular senescence. Mechanistic studies revealed that cardamonin reduced ROS accumulation and suppressed mTOR phosphorylation. Notably, MHY1485-mediated activation of the mTOR pathway reversed senescence and fibrotic phenotypes, providing reciprocal validation of the target mechanism. In conclusion, cardamonin mitigates cellular senescence by targeting the mTOR-ROS axis, offering a promising therapeutic strategy for anti-aging interventions and the treatment of idiopathic pulmonary fibrosis (IPF).