Multi-omics uncovers formononetin's protection against antibiotic-induced liver injury via microbiota-metabolite-transcriptome coordination

Tetracycline antibiotics (TETs) cause hepatotoxicity via gut-liver axis disruption and oxidative stress. This study evaluated whether formononetin (FMN), a natural isoflavone, alleviates TET-induced liver injury through microbiota-metabolite-transcriptome coordination. Using network toxicology and pharmacology, we predicted key targets and pathways involved. A mouse model of TET-induced injury (150 mg/kg, 3 days) was treated with FMN (50 or 100 mg/kg) for 14 days. Multi-omics analyses revealed that FMN attenuated hepatic steatosis, reduced ALT/AST levels, enhanced gut microbial diversity (enriched Limosilactobacillus), and restored intestinal barrier integrity. Mechanistically, FMN activated the Nrf2/HO-1 antioxidant pathway, modulated PPARα/CYP2E1-dependent lipid metabolism, and inhibited ferroptosis. FMN demonstrates multi-target hepatoprotection by coordinating microbiota restoration, oxidative stress reduction, and metabolic reprogramming, supporting its potential as a dietary intervention against antibiotic-related liver damage.