Network toxicology and machine learning revealed the association between non-alcoholic fatty liver disease induced by polystyrene nanoplastics

This study systematically evaluated the hepatotoxic potential of common microplastic monomers using ADMETlab 3.0, identifying styrene (PubChem ID: 7501) as exhibiting significant hepatic toxicity. Comprehensive analysis of the CTD (Comparative Toxicogenomics Database) revealed a strong association between styrene exposure and NAFLD (Non-alcoholic fatty liver disease). We integrated target prediction data from CTD, STITCH and SwissTargetPrediction databases yielding 558 candidate targets, which were cross-referenced with 2018 NAFLD related genes from GeneCards and OMIM to identify 158 overlapping genes. Applying the NCBI GEO datasets GSE48452, GSE126848, GSE89632, and GSE63067, comprising 98 control samples and 113 NAFLD samples, we trained the model using LASSO regression, random forest, and SVM-RFE algorithms to identify the core genes HSPD1 and NFE2L2. These findings were validated in the GSE135251 cohort, which included 10 control cases and 206 NAFLD cases. In vitro experiments showed 100 nm polystyrene nanoparticles (PS-NPs) at 200 μg/mL for HepG2 and 100 μg/mL for AML12 cells significantly increased lipid accumulation by Oil Red O staining. Western blot revealed dose-dependent changes in lipid metabolism regulators ACC1, FASN and CPT1A. Preliminary RT-qPCR and immunofluorescence studies indicate that PS-NPs can upregulate HSPD1 and NFE2L2. These findings suggest that PS-NPs promote hepatic lipid accumulation by regulating HSPD1 and NFE2L2, offering new insights into the pathogenesis of environmentally induced NAFLD.