Neutrophils, essential cells of the innate immune system, play dual roles in the defense of host and disease pathogenesis. Neutrophils crosstalk with endothelial cells to maintain homeostasis via mechanisms such as phagocytosis, neutrophil extracellular trap (NET), and reactive oxygen species (ROS) generation, excessive neutrophil activity contributes to inflammation and tissue damage in various diseases, including autoimmune disorders and cancer. This study investigates the effects of HUVEC-derived exosomes (H-Exo) on neutrophil function. H-Exo were isolated, and characterized using TEM, DLS, and flow cytometry, and their effects on neutrophils were evaluated. Our results demonstrate that H-Exo significantly reduces neutrophil ROS production, as confirmed by the Nitroblue Tetrazolium (NBT) test, and decreases neutrophil yeast killing activity. These findings highlight the potential of H-Exo in managing diseases associated with excessive neutrophil activation and oxidative stress. In conclusion, H-Exo represents a promising therapeutic avenue for conditions characterized by neutrophil-induced inflammation.