Disposable Plastic-Alcohol Leachates as Emerging Neurotoxicants: Evidence for the miR-330-3p/Acsl1 Pathway in Cognitive Performance.

Chronic alcohol consumption impairs cognitive performance via neurotoxic effects. Recent evidence suggests that chemical leachates from disposable cups may exacerbate this toxicity. Pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were employed to characterize alcohol-induced leachates from disposable cups. Male rats were then administered the respective leachates or alcohol alone via oral gavage every 48 hours for 8 weeks. Cognitive performance was evaluated through behavioral testing, and hippocampal morphology was examined using hematoxylin-eosin (H&E) staining and transmission electron microscopy (TEM). Integrative analyses of the miRNA-mRNA regulatory network and protein-protein interaction (PPI) network identified the rno-miR-330-3p/Acsl1 axis as a key regulatory pathway, which was subsequently validated by quantitative real-time polymerase chain reaction (qRT-PCR). Py-GC/MS and LC-MS/MS analysis revealed that compounds derived from microplastics (MPs) and per- and polyfluoroalkyl substances (PFAS) migrated into alcohol, and subsequent exposure to these leachates induced hippocampal structural alterations, including CA3 pyramidal neuron shrinkage and morphological abnormalities, Blood-Brain Barrier (BBB) disruption, and mitochondrial swelling, accompanied by cognitive performance changes. These findings demonstrate that disposable plastic cup leachates exacerbate alcohol-induced neurotoxicity by dysregulating the rno-miR-330-3p/Acsl1 signaling axis. Nonstandard Abbreviations and Acronyms: BBB = Blood-Brain Barrier; MPs = microplastics; PVC = Polyvinyl Chloride; PA66 = Polyamide 66; PFAS = Per- and Polyfluoroalkyl Substances; PFUnDA = Perfluoroundecanoic Acid; PFBS = Perfluorobutane Sulfonic Acid; NOR = Novel Object Recognition; MWMT = Morris Water Maze Test; H&E = Hematoxylin and Eosin; TEM = Transmission Electron Microscopy; DE = Differential Expression; PPI = Protein-Protein Interaction; qRT-PCR = Quantitative Real-Time Polymerase Chain Reaction; GO = Gene Ontology; KEGG = Kyoto Encyclopedia of Genes and Genomes; Py-GC/MS = Pyrolysis-Gas Chromatography/Mass Spectrometry.