Puerarin‐Loaded Macrophage‐Derived Exosomes for Experimental Atherosclerosis

ABSTRACT Atherosclerotic cardiovascular disease (AS) remains a leading global health concern. In this study, we investigated a macrophage‐derived exosome‐based delivery system for puerarin (Pue). The isolated vesicles were characterized by transmission electron microscopy, particle‐size and zeta‐potential analysis, and Western blot validation of extracellular vesicle‐associated markers, including CD63, CD9, CD81, TSG101, and Alix, together with Calnexin as an endoplasmic‐reticulum‐associated negative/depletion marker. In vitro experiments showed that exosome‐based delivery enhanced cellular uptake of puerarin and was associated with reduced oxidative‐stress‐related signals, decreased macrophage migratory activity, and reduced foam‐cell‐related readouts under the tested conditions. In vivo studies using an ApoE−/− mouse model showed that Pue–Exos reduced aortic plaque burden, promoted collagen deposition, and improved serum lipid‐related indices. No obvious histopathological abnormalities were observed in major organs after treatment, suggesting preliminary in vivo tolerability. Mechanistic evaluation in C57BL/6 mice showed that treatment was associated with increased expression of HIF‐1α, NF‐κB, IL‐6, IL‐1β, CD68, and α‐SMA under the present experimental conditions. Therefore, these findings are more appropriately interpreted as inflammation‐associated signaling changes and remodeling‐related responses, rather than definitive evidence of a uniformly suppressed inflammatory state or a confirmed anti‐atherosclerotic mechanism. Taken together, these results suggest that macrophage‐derived exosome‐based delivery may improve the performance of puerarin in atherosclerosis‐related experimental settings, although the underlying mechanism requires further clarification.