Integrated in vitro and in vivo omics analyses reveal lipid homeostasis disruption induced by widely used antimicrobial agent zinc pyrithione.

Zinc Pyrithione (ZnPT), a widely used antimicrobial agent, has been increasingly detected in environmental matrices, however its toxicological effects remain insufficiently characterized. Here, we employed an integrated in vitro and in vivo approach to systematically evaluate the impact of ZnPT on lipid homeostasis at environmentally relevant concentrations. In vitro studies using HepG2 cells demonstrated ZnPT exposure significantly perturbed intracellular lipid balance. Nile Red staining revealed a dose-dependent decrease in neutral lipid content, detectable even at the lowest concentrations (≥ 4.0 nmol/L). Lipidomic profiling further revealed substantial alterations across multiple lipid subclasses, including the glycerophospholipids, sphingolipids, glycerolipids, and fatty acids. Notably, increased levels of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were observed. Transcriptomic analysis identified widespread dysregulation of lipid metabolism-related genes, with significant enrichment in pathways associated with lipid biosynthesis, transport, and degradation. Complementary in vivo experiments in zebrafish embryos further corroborated the in vitro findings. A significant reduction in total triglyceride (TG) content (-12.5 %) was observed following ZnPT exposure. Additionally, multivariate analysis (OPLS-DA) and integrated gene-lipid network analysis confirmed key associations between differentially expressed genes and altered lipid species. Collectively, these findings provide compelling evidence of ZnPT-induced disruption of lipid homeostasis at low concentrations and offer important new insights for environmental risk assessments of novel antimicrobial agents.