Mechanistic insights into compatibility-based detoxification of Fengshi Gutong Capsules through network toxicology and in vivo validation

ETHNOPHARMACOLOGICAL RELEVANCE: Aconitum is a crucial medicinal herb widely used in the treatment of rheumatic diseases; however, its clinical application is restricted due to its inherent toxicity. In traditional Chinese medicine, the efficacy of Aconitum is enhanced and its toxicity reduced through the principle of "herbal compatibility." Fengshi Goutong Capsules (FSGTC) represent a classical prescription exemplifying this compatibility-based detoxification strategy. Although FSGTC has demonstrated significant therapeutic efficacy with minimal adverse effects, the underlying mechanisms responsible for its toxicity attenuation remain unclear. OBJECTIVE: This study aimed to identify the active components in FSGTC responsible for mitigating Aconitum-induced toxicity and to elucidate the underlying molecular mechanisms. MATERIALS AND METHODS: Network toxicology analysis was employed to screen the key toxic components of Aconitum in FSGTC, as well as the active components and potential targets involved in toxicity attenuation. Molecular docking and molecular dynamics simulations were performed to validate the binding affinities between active components and core targets. Key detoxification-related signaling pathways were identified through Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. These predictions were subsequently validated through in vivo experiments, including behavioral assessments, organ-to-brain ratio measurements, evaluation of organ function biomarkers, histopathological examination, and analysis of protein expression levels of key pathway targets. RESULTS: Network toxicology analysis identified hypaconitine as the principal toxic component of Aconitum in the FSGTC formula, while quercetin from the compatible herbal components was identified as the primary detoxifying compound. ATP-binding cassette subfamily B member 1 (ABCB1) and ATP-binding cassette subfamily G member 2 (ABCG2) were determined to be the core targets mediating quercetin-induced detoxification. Computational simulations confirmed the stable binding of quercetin to these targets. KEGG pathway enrichment analysis revealed that the cytochrome P450 (CYP450) metabolic pathway played a central role in quercetin-mediated toxicity attenuation. Animal experiments demonstrated that quercetin significantly ameliorated hypaconitine-induced behavioral abnormalities, changes in organ-to-brain ratios, and functional as well as structural damage to the heart, liver, kidneys, and brain in rats. Immunohistochemical analysis further revealed that quercetin promoted hypaconitine metabolism by restoring the activity of the CYP450 pathway and regulating the expression of downstream transporters, ABCB1 and ABCG2. CONCLUSION: Quercetin in FSGTC alleviates the cardiotoxicity, hepatotoxicity, nephrotoxicity, and neurotoxicity induced by hypaconitine. The underlying mechanism involves the restoration of the activity of the CYP450 pathway and regulation of downstream transporters (ABCB1/ABCG2), thereby promoting hypaconitine metabolism and promoting detoxification.