Integrated network toxicology and molecular docking strategy elucidate ferroptosis mechanisms underlying acetyl tributyl citrate induced cardiotoxicity

This study aims to elucidate the cardiotoxic effects of acetyl tributyl citrate (ATBC) and unravel the underlying mechanisms governing the impact of this environmental pollutant on cardiac-related pathological processes. Ferroptosis is emerging as a critical cell death pathway, playing a pivotal role in adverse outcomes associated with exposure to environmental pollutants and toxins. Utilizing ChEMBL, STITCH, GeneCards, and OMIM databases, we first identified a comprehensive list of 269 potential targets closely linked to ATBC and cardiotoxicity. From the FerrDb v2 and GeneCards databases, we obtained 469 ferroptosis-related genes. Intersection analysis yielded 25 shared genes among these three sets. Following rigorous refinement using the STRING database, the cytoHubba plugin, and Cytoscape software, we pinpointed nine core targets: Caspase 8, TNF, SRC, IDH2, MAPK14, KRAS, IDH1, ATM, and KEAP1. Subsequently, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that the core targets implicated in ATBC-induced cardiotoxicity were significantly enriched in ferroptosis-related pathways, such as Chemical carcinogenesis-reactive oxygen species. Further narrowing of core targets was achieved through MCODE and CytoHubba clustering analyses, yielding three pivotal targets: Caspase 8, IDH2, and MAPK14. Furthermore, molecular docking simulations using AutoDock confirmed strong binding affinity between ATBC and these core targets, providing deeper insights into their interaction mechanism. Then, to investigate the effect of ATBC on cellular ferroptosis, we conducted in vitro experiments using cardiomyocytes exposed to relevant concentrations. Our findings demonstrated that low-dose ATBC significantly impaired cardiomyocyte viability. Concurrently, we observed altered expression of three pivotal targets and significant activation of ferroptosis in cardiomyocytes. This indicates that ATBC can induce myocardial injury at the cellular level via ferroptosis mechanisms. Collectively, our results provide a theoretical framework for understanding the complex mechanisms mediating ATBC-induced myocardial damage. They also offer crucial insights for developing preventive and therapeutic strategies targeting cardiac disorders potentially arising from exposure to ATBC-containing plastic products or environmental ATBC contamination.