Integration of multi-omics and machine learning to reveal the anti-osteoporosis effects of Hederagenin by inhibiting lipid peroxidation via the GPX4/p-STAT1/PTGS2 axis.

Osteoporosis (OP) is a multifactorial disease, leading to abnormal bone remodeling, requiring targeted therapeutic interventions. This study investigated the pharmacological effects of Hederagenin (Hed), a triterpenoid saponin, in ovariectomized (OVX) mice. Hed treatment significantly ameliorated bone loss by improving trabecular microstructure and reducing osteoclast formation and activity. Integrated metabolomics and network pharmacology revealed that Hed primarily restored glycerophospholipid metabolic homeostasis and suppressed oxidative stress. Crucially, biochemical assays demonstrated that Hed treatment significantly elevated superoxide dismutase (SOD) and glutathione (GSH) levels, while reducing malondialdehyde (MDA) and oxidized low-density lipoprotein (ox-LDL) accumulation. Concurrently, quantitative reverse transcription PCR (qRT-PCR) and Western blot analyses confirmed that Hed downregulated the expression of lipogenic genes and the key enzyme lysophosphatidylcholine acyltransferase 1 (Lpcat1). Machine learning-based transcriptomics identified prostaglandin-endoperoxide synthase 2 (PTGS2) as a pivotal target associated with lipid peroxidation, with signal transducer and activator of transcription 1 (STAT1) acting as its upstream transcriptional regulator. Subsequent experiments revealed that Hed upregulated glutathione peroxidase 4 (GPX4) expression, thereby reducing phosphorylated STAT1 (p-STAT1) levels and suppressing PTGS2 levels. Mechanistically, Hed modulates the GPX4/p-STAT1/PTGS2 axis to inhibit lipid peroxidation, thereby alleviating OVX-induced bone loss. These findings unveil Hed as a promising therapeutic candidate and provide novel mechanistic insights into the treatment of osteoporosis.