Oxidative stress-mediated impairment of human trophoblast cell proliferation by zinc pyrithione exposure

Zinc pyrithione (ZPT), a broad-spectrum antimicrobial agent widely used in anti-dandruff shampoos and antifouling coatings, has an unclear toxic effect on embryonic trophoblast cells. To systematically evaluate the toxicological impact of ZPT on human trophoblast cell line JEG-3 and its underlying mechanisms, cells were treated with 90 nM ZPT for 72 h. A series of assays, including Cell Counting Kit-8(CCK-8), flow cytometry, wound healing, and Transwell, were performed to assess cell proliferation, apoptosis, migration, and invasion. Intracellular reactive oxygen species (ROS) levels and DNA damage were assessed using the 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) probe and γ-H2AX immunofluorescence, respectively. Transcriptome sequencing and Gene Ontology(GO) enrichment analysis were also performed. The results indicated that ZPT significantly inhibited cell proliferation, migration, and invasion, induced late-stage apoptosis, and increased ROS levels and DNA damage. RNA sequencing (RNA-seq) identified 1020 differentially expressed genes, suggesting an upregulation in autophagy and mitochondrial apoptosis pathways, and a significant downregulation in glycolysis, NAD⁺ regeneration, and hypoxia response pathways. Quantitative real-time polymerase chain reaction (qPCR) validation further confirmed the upregulation of key stress- and autophagy-related genes (NUPR1, SQSTM1) and the downregulation of genes involved in trophoblast function and mitochondrial quality control (BMP4, BNIP3, BNIP3L). These in vitro findings suggest that ZPT may impair trophoblast function through mechanisms involving oxidative stress, DNA damage, and perturbations in mitochondrial apoptosis/autophagy and energy metabolism.