Objective: To investigate the mechanism of Dendrobium-derived exosome-like vesicles against atherosclerosis by regulating macrophage polarization.Methods: ①Dendrobium-derived exosome-like vesicles were isolated by density gradient centrifugation, and characterized and identified via transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The uptake of Dendrobium-derived exosome-like vesicles by RAW264.7 macrophages was observed under confocal laser scanning microscopy. ②The chemical components of Dendrobium-derived exosome-like vesicles were analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS). Combined with network pharmacology, the core targets and key pathways of Dendrobium-derived exosome-like vesicles in regulating macrophage polarization for atherosclerosis treatment were screened. ③ In vitro, the M1-type macrophage polarization model was established by LPS + IFN-γ induction. Different concentrations of Dendrobium-derived exosome-like vesicles were set for intervention. The expression levels of inflammatory factors were detected by RT-qPCR and Western Blot. ④ In vivo, the atherosclerosis model was constructed using ApoE?/? mice. After intraperitoneal injection of Dendrobium-derived exosome-like vesicles, the effects of DDNVs on atherosclerotic plaque area, blood lipid levels, inflammatory factors, and macrophage polarization-related signaling pathways were evaluated by oil red O staining, hematoxylin-eosin (HE) staining, blood lipid detection, ELISA, RT-qPCR, immunofluorescence, and immunohistochemistry.Results: ①Dendrobium-derived exosome-like vesicles exhibited a typical cup-shaped morphology with an average particle size of 154.2 nm, and could be internalized by macrophages. ② A total of 316 compounds were identified by HPLC-MS, among which 209 compounds had a comprehensive score> 60.Network pharmacology analysis identified 128 common targets of Dendrobium-derived exosome-like vesicles, atherosclerosis, and macrophage polarization, which were enriched in signaling pathways related to inflammatory response regulation. ③In vitro experiments showed that Dendrobium-derived exosome-like vesicles inhibited M1-type macrophage polarization in a concentration-dependent manner, reduced the expression of pro-inflammatory factors IL-1β and iNOS, and increased the expression of anti-inflammatory factor IL-10. ④In vivo experiments demonstrated that Dendrobium-derived exosome-like vesicles significantly reduced the aortic plaque area, improved blood lipid levels, alleviated aortic pathological damage, decreased serum IL-1β level and increased serum IL-10 level in atherosclerotic mice. Meanwhile, they decreased the proportion of M1-type macrophages and increased the proportion of M2-type macrophages in the aortic root, as well as inhibited the activation of the JAK2/STAT3 signaling pathway.Conclusion: Dendrobium-derived exosome-like vesicles exert anti-atherosclerotic effects by regulating the balance of macrophage polarization, alleviating inflammatory responses, and improving lipid metabolism, whose mechanism may be associated with the inhibition of the JAK2/STAT3 signaling pathway.