Enhanced reproductive toxicity following co-exposure to disposable container leachates and alcohol involves proteomic identification.

OBJECTIVE: To investigate the enhanced reproductive effects of co-exposure to disposable container leachates and alcohol in rats, with particular emphasis on elucidating the underlying molecular mechanisms by identifying core regulatory proteins and pathways. METHODS: Pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS), gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to characterize microplastics (MPs), phthalate esters (PAEs), per- and polyfluoroalkyl substances (PFAS) and bisphenol A (BPA) in disposable container leachates. Animal experiments were conducted to evaluate testicular histology, blood-testis barrier integrity, sperm quality, and sex hormone levels, combined with proteomic analysis, bioinformatics analysis, and parallel reaction monitoring (PRM) to elucidate the relevant molecular mechanism. RESULTS: Two MPs, polyvinyl chloride (PVC) and polyamide 6 (PA66), along with three PFAS compounds, Perfluorooctanoic acid (PFOA), perfluoroundecanoic acid (PUnDA) and perfluorobutanesulfonic acid (PFBS), were detected in the leachates, confirming their migration into Chinese baijiu.Co-exposure resulted in a significant reduction in body weight gain rate, more severe testicular seminiferous tubule damage, disruption of the blood-testis barrier, and greater declines in sperm quality, accompanied by exacerbated imbalances in testosterone (T) and follicle-stimulating hormone levels. Proteomics identified three key core proteins, Hpx (emopexin), Ahsg (lpha-2-HS-glycoprotein), Ttr (Transthyretin), which were validated by PRM. CONCLUSION: Co-exposure to cup leachates and alcohol enhances reproductive toxicity, with PC-AL disrupting endoplasmic reticulum and PLC-AL damaging mitochondria and downregulating Hpx, Ahsg, and Ttr, which disrupt pathways and immune microenvironment, impairing reproductive structure.