Broccoli-derived nanovesicles inhibited LPS-induced inflammatory response in mouse macrophage RAW264 cells

We prepared nanovesicles (NVs) derived from broccoli using ultracentrifugation and evaluated their anti-inflammatory properties. Two distinct NV populations were isolated as precipitates from broccoli homogenates following centrifugation at 20 000 × g and 200 000 × g. These NVs contained RNAs, proteins, isothiocyanates, and chlorophylls. Dynamic light scattering analysis confirmed their nanoscale size. The NVs were internalized by RAW264 cells and significantly inhibited nitric oxide production and NF-κB pathway activation under lipopolysaccharide (LPS) stimulation. Comprehensive analysis of inflammatory cytokine expression revealed strong suppression of interleukin-6 (IL-6) by both NV types, which was further validated by ELISA. Additionally, IL-1β and tumor necrosis factor-α production were also reduced. Notably, the anti-inflammatory effects were partially attributed to small RNAs (<200 nt) present within the NVs. Collectively, these findings suggest that broccoli-derived NVs possess potent anti-inflammatory activity.