Mechanistic insights into PI3K/Akt-MMP9 signaling in Crotonis Fructus-induced reproductive toxicity via integrated network toxicology and multi-model validation

Despite the widespread use of Crotonis Fructus (derived from Croton tiglium L.) in traditional medicine, its reproductive toxicity remains poorly characterized. Here, we investigated its potential mechanisms using Caenorhabditis elegans, network toxicology, molecular docking, and multi-model validation. Exposure to croton extract (CE) impaired reproductive capacity and shortened lifespan in C. elegans. Network toxicology identified 21 putative toxic compounds and 88 female reproductive dysfunction-related targets. Molecular docking revealed strong binding affinities of phorbol diesters, among others (key Crotonis Fructus components) to critical targets, including AKT1 and MMP9. In vitro (RAW264.7 cells) and in vivo (SD rats) experiments demonstrated CE-induced dysregulation of PI3K/Akt signaling pathway genes. Our findings establish that Crotonis Fructus induces female reproductive toxicity by regulating the release of inflammatory factors and activating the PI3K/Akt pathway, providing critical insights for refining its clinical safety guidelines.