Multi-omics decoding of Phalloidin hepatotoxicity: network toxicology-guided identification of FoxO/PLD/cAMP signaling targets.

Phalloidin is a lethal toxin found in highly toxic mushrooms of the genus Amanita; it causes severe liver injury, yet its molecular mechanism has not been systematically elucidated. In this study, Male Kunming mice received a single intraperitoneal injection of Phalloidin (0.3 mg/kg and 0.6 mg/kg). we predicted Phalloidin targets by network toxicology and explored its toxicity mechanism by combining metabolomics and proteomics in the livers of Phalloidin-toxicized mice. Metabolomic and proteomic profiling revealed that Phalloidin broadly perturbed hepatic metabolism. Specifically, it down-regulated nucleotide metabolism, coenzyme A synthesis and phospholipid metabolism, and reduced the DNA-replication-related MCM family proteins. Concomitantly, the lipid-metabolism proteins Hsd17b6 and Dhcr24 were aberrantly expressed. These alterations collectively promoted hepatocyte death. Integrative multi-omics analysis further identified abnormal expression of ten core regulatory targets, including the cell-cycle checkpoint protein Cdkn1b, the MAPK signaling proteins Mapk14 and Map2k2, and the insulin signaling components Insr and Igf2r. Functional enrichment indicated that Phalloidin synergistically induces hepatic injury by interfering with three key pathways: the FoxO, phospholipase D and cAMP signaling cascades. Molecular-docking simulations confirmed high-affinity binding between Phalloidin and these core targets, providing direct evidence of their physical interaction. Collectively, this study systematically delineates the molecular network underlying Phalloidin-induced liver injury and proposes these pathways as potential therapeutic targets for Amanita mushroom poisoning.