Integrating transcriptomics and phenotypic profiling reveals caffeic acid-induced multi-organ developmental toxicity in zebrafish.

Caffeic acid is widely used in food and pharmaceuticals, raising concerns about its aquatic ecotoxicity. Here, we evaluated developmental toxicity in zebrafish embryos acutely exposed to caffeic acid. At 120 hpf, the LC₅₀ was 134.3 μg/mL. Subsequent exposures at 0, 40, 80, and 120 μg/mL dose-dependently increased mortality, delayed hatching, reduced growth, and induced cardiac, hepatic, and neurobehavioral abnormalities. Molecular assays revealed dysregulation of inflammation-related genes and suppressed expression of caspase3/9 with a decreased bax/bcl2 ratio, indicating an inflammatory response and inhibition of apoptosis. Transcriptomic profiling at 80 μg/mL identified 287 differentially expressed genes enriched in immune, cellular, and signal transduction pathways; protein-protein interaction analysis further suggested possible synergistic roles in DNA repair and cell cycle regulation. Collectively, our results demonstrate that caffeic acid causes multi-organ developmental toxicity at phenotypic and gene-expression levels. The findings provide a theoretical basis for understanding the toxicological mechanisms of caffeic acid and contribute to its environmental safety assessment.