Integrated network toxicology and multiomics analyses reveal the mechanisms underlying trimethyltin chloride-induced epilepsy and the ameliorative effects of astaxanthin via NFKB1 and IL6.

Trimethyltin chloride (TMT) is a ubiquitous neurotoxic organotin pollutant that induces epilepsy, while the natural marine product astaxanthin (AST) exerts potent antioxidant, anti-inflammatory and neuroprotective effects. However, the causal molecular mechanisms of TMT-induced epilepsy and the targeted intervention mechanism of AST remain unclear. In this work, we adopted a multidimensional analytical framework encompassing an National Health and Nutrition Examination Survey (NHANES) population cohort analysis, network toxicology, machine learning, single-cell RNA sequencing, Mendelian randomization (MR), virtual gene knockout, molecular docking, molecular dynamics (MD) simulations and in vivo animal experiments to systematically identify and verify the core targets and candidate therapeutic compounds for TMT-induced epilepsy. The population analysis confirmed a positive dose-response correlation between tin exposure and the epilepsy risk. Network toxicology identified 157 overlapping target genes linked to both TMT exposure and epilepsy. Protein-protein interaction (PPI) network analysis together with a systematic evaluation of 77 machine learning models further revealed ten core hub genes. Furthermore, MR verified NFKB1 and IL6 as core causal pathogenic genes. Virtual knockout of NFKB1 and IL-6 combined with functional enrichment analysis yielded three shared downstream genes: SEMA5A, PTPRZ1, and GRM7. Molecular docking and dynamics characterized AST and key genes interactions. Consistent with these in silico findings, the results of in vivo animal experiments demonstrated that AST ameliorates TMT-induced epileptic phenotypes by suppressing NFKB1 and IL6 expression. Collectively, this multiomics study reveals the dominant pathogenic roles of NFKB1/IL-6 in TMT-induced epilepsy and underscores AST as a promising therapeutic candidate for counteracting TMT-mediated epileptic injury.