Abstract: Plant exosome-like nanovesicles (ELNVs), as inherent components of plants, are nanoscale vesicles secreted by plant cells. They are inherently rich in bioactive compounds, and their vesicular structure makes them ideal “green” drug delivery vehicles. To overcome the limitations of single-source plant ELNVs, this study employed membrane fusion technology to combine tea ELNVs and Citrus reticulata ‘Chachiensis’ ELNVs for the first time, creating novel fusogenic nanovesicles. We subsequently characterized the physicochemical properties of novel fusogenic nanovesicles, elucidated their fundamental composition, assessed their stabilities in simulated digestive fluids, and evaluated their antioxidant, anti-inflammatory and anti-tumor activities through cellular assays. The results indicate that the novel fusogenic nanovesicles are a reconstituted liposome integrating multiple functional components, including tea polyphenols (catechins), hesperetin, hesperidin, nobiletin, quercetin, active phospholipids and proteins. They exhibit a spherical morphology with a phospholipid bilayer structure, demonstrating good dispersibility with uniform particle size. Additionally, the novel fusogenic nanovesicles demonstrate excellent stability in both simulated gastric and intestinal fluids. Cell-based experimental results show that the novel fusogenic nanovesicles significantly alleviated lipopolysaccharide (LPS)-induced oxidative stress damage and inflammatory responses in RAW264.7 cells, while exhibiting superior antitumor efficacy. The findings of this study not only provided novel insights into the application of natural plant ELNVs, but also established a scientific foundation for developing innovative multifunctional oral nutraceuticals.

Key words: plant exosome-like nanovesicles (plant ELNVs), reconstituted liposomes, anti-inflammatory, antioxidant, antitumor