Mechanisms of Etomidate Addiction and Neuro-Hepatic Toxicity: Molecular Pathway Analysis Based on Multi-Omics Integration

Abstract Etomidate has been linked to addictive behaviors and neurotoxic and hepatotoxic effects. We established a mouse exposure model across graded etomidate doses and assessed reward using conditioned place preference (CPP). Proteomic and metabolomic profiling of cerebral tissue and right hepatic lobe tissue was performed using data-independent acquisition (DIA) liquid chromatography–tandem mass spectrometry and quadrupole time-of-flight mass spectrometry, respectively; integrated multi-omics candidates were verified by parallel reaction monitoring (PRM). Both low- and high-dose etomidate significantly increased CPP scores, indicating robust rewarding properties. Neuronal and hepatic lesions increased with dose, with the prefrontal cortex and ventral tegmental area showing the greatest brain vulnerability. Integrated proteomic and metabolomic analyses indicated shared biotransformation- and stress-response features across brain and liver, with brain changes mainly involving xenobiotic metabolism, oxidative stress-related detoxification, and synaptic signaling and cAMP–RAS signaling, and liver changes mainly involving cytochrome P450-mediated biotransformation, lipid and fatty-acid metabolism, and lethal-stage catabolic remodeling. Metabolomics showed dose-stratified pathway disruption: low-dose exposure affected aminoacyl-tRNA and amino acid biosynthesis, high-dose exposure perturbed amino acid and carbon metabolism, and lethal doses altered purine metabolism and glutamatergic pathways. The main innovation of this study lies in establishing a dose-stratified mouse model that integrates CPP behavior, brain and liver pathological injury, and cross-organ proteomic–metabolomic profiling, thereby extending existing evidence of etomidate-related reward and toxicity and delineating a combined molecular profile of etomidate-induced reward-related behavior and organ toxicity.