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.