Integrated network toxicology, molecular docking, molecular dynamics simulation, and experimental verification to...

Fructus Meliae Toosendan (FMT) has been traditionally used in Chinese medicine, yet its mechanisms of its hepatotoxicity, as well as the detoxification process following processing, are still not fully elucidateed. This study aimed to investigate the hepatotoxic components and targets from FMT and elucidate both its mechanism of hepatotoxicity and the detoxification mechanism induced by processing. We employed a combination of network pharmacology, molecular docking, molecular dynamics simulations and in vitro experiments to explore the potential hepatotoxic targets and mechnisms of FMT. A total of 12 toxicity components and 71 potential hepatotoxicity targets of FMT were identified. Protein-protein interaction (PPI) network analysis identified the top six core potential targets, three of which possess suitable crystal structures for molecular docking. These include meliasenin B-HSD17B4 (ΔG = -7.40 kcal mol^-1) and meliasenin B-HMGCR (-7.32 kcal mol^-1), melianone-HSD17B4 (-8.19 kcal mol^-1) and melianone-SOD2 (-8.51 kcal mol^-1). toosendanin-HMGCR (-7.58 kcal mol^-1). Additionally, 185 Gene Ontology (GO) terms and 72 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were identified. The three major toxicity ingredients and three core targets mentioned above were subjected to molecular docking analysis using AutoDock Vina. Molecular dynamic simulations confirmed a stable interaction between the meliasenin B, melianone and toosendanin and HSD17B4, HMGCR and SOD2 ligand system. It indicated that melianone, toosendanin and isotoosendanin significantly reduced cell viability in a dose-dependent manner in Human hepatoma cells HepG2 and Human normal liver cell line L02 hepatocytes in vitro experiments. Processing FMT enhances viability in both HepG2 and L02 cells while mitigating its hepatotoxic effects. The hepatotoxicity-reducing effect of processing FMT may involve the PI3K/AKT/FOXO signaling pathway. This study suggests that melianone, toosendanin, isotoosendanin and meliasenin B may be hepatotoxic components of FMT, and the detoxification mechanism of processing FMT may be related to the PI3K/AKT/FOXO signaling pathway, warranting further investigation into their safety profiles in clinical applications.