The adverse effects of paromomycin on early cardiovascular development in zebrafish (Danio rerio)

• First systematic confirmation of the early cardiovascular developmental toxicity of paromomycin: For the first time, using zebrafish embryos as a model, it was clarified that paromomycin, at environmentally relevant exposure concentrations, can induce cardiovascular developmental damage in early embryos in a dose- and time-dependent manner. • Innovatively integrating network toxicology, molecular docking, and transcriptomics, this study overcomes the limitations of single-method approaches and establishes a comprehensive mechanistic framework from toxic phenotypes to targets and pathways. • It not only clarifies the molecular mechanism of paromomycin-induced cardiovascular toxicity, but also provides direct experimental evidence for the safe use and ecological risk assessment of paromomycin based on the toxicity results at environmentally relevant concentrations, thereby bridging the gap between basic research and practical application. Paromomycin, a broad-spectrum aminoglycoside antibiotic, is widely used in both clinical and agricultural fields. However, its environmental impact and potential developmental toxicity have raised growing concerns. This study systematically evaluates the adverse effects of paromomycin on early cardiovascular development in zebrafish embryos from multiple biological perspectives. Zebrafish embryos were exposed to environmental concentrations of paromomycin (0, 0.3, 3, 30, and 300 μg/mL) for 5 days. Comprehensive assessments were then performed through morphological observations, cardiovascular phenotype analysis, behavioral evaluations, and integrated analysis. The results revealed that exposure to paromomycin induced dose dependent developmental damage, including decreased survival rates, delayed hatching, and increased malformation rates. Detailed cardiovascular assessments confirmed significant abnormalities in both cardiac function and vascular development. Molecular level investigations further elucidated the underlying mechanisms, showing that paromomycin exposure elevated reactive oxygen species levels, triggering oxidative stress, inducing apoptosis in the pericardial region, and disrupting the expression of key cardiovascular developmental genes. Integrating network toxicology, molecular docking, and transcriptomic analyses, this study identified six key signaling pathways involved in paromomycin induced cardiovascular toxicity. These findings provide a comprehensive mechanistic framework for understanding the developmental cardiovascular toxicity of paromomycin at environmentally relevant concentrations and offer valuable insights for the safe application and risk management of this widely used antibiotic.