Multifunctional Biological Activity Assessment of Plant-Derived Nanovesicles from Arugula Leaves: In Vitro and In Vivo Studies

Plant-derived vesicles (PDVs) represent an emerging class of naturally bioformulated nanocarriers with potential nutraceutical and therapeutic applications. In this study, the multifunctional biological activity of PDVs obtained from <i>Eruca sativa</i> leaves (arugula leaf vesicles, ALVs) was investigated both in vitro and in vivo. In differentiated Caco-2 and HepG2 cells, ALVs exhibited significant antioxidant activity, being rich in polyphenols and organic acids, by reducing intracellular reactive oxygen species (ROS) and modulating key metabolic regulators. ALVs upregulated SREBP-2, LDLR, and phosphorylated AMPK and Akt, leading to enhanced LDL and glucose uptake, while downregulating FASN and PPAR-γ, thereby reducing lipid accumulation. In mice fed a high-fat and high-fructose (HFHF) diet, ALV supplementation improved glucose tolerance and decreased total cholesterol, LDL, and hepatic injury biomarkers (ALT, AST, and LDH) without inducing toxicity. These findings demonstrate that ALVs exert hypocholesterolemic, hypoglycemic, and lipid-lowering effects through coordinated modulation of AMPK/Akt pathways. Overall, ALVs emerge as safe, multifunctional nanovesicles capable of counteracting oxidative stress and metabolic dysfunction, highlighting their potential as innovative bioactive ingredients for functional foods or nutraceutical formulations targeting metabolic syndrome.