090 | Activation of Sirtuin-1 by Cipollotto Nocerino leaf extract protects HepG2 cells from lipotoxicity

Vincenzo Vestuto¹, Maria Rosaria Miranda¹, Giovanna Aquino², Tania Ciaglia¹, Rodolfo Maria Abbate¹, Giuseppina Amodio³, Manuela Giovanna Basilicata⁴, Mario Felice Tecce¹, Luca Rastrelli¹, Pietro Campiglia¹, Michele Manfra⁵, Giacomo Pepe¹ | ¹Department of Pharmacy, University of Salerno, Italy; ²AREA Science Park, Laboratorio di Multi-omica Area Sud LAAS, Baronissi [SA], Italy; ³Department of Medicine, Surgery and Dentistry "Scuola Medica Salernitana", University of Salerno, Baronissi [SA], Italy; ⁴Department of Advanced Medical and Surgical Sciences, University of Campania "Luigi Vanvitelli", Naples, Italy; ⁵Department of Science, University of Basilicata, Potenza, Italy.

Mediterranean agri-food by-products offer nutraceuticals with potential benefits in metabolic disorders. The Cipollotto Nocerino (CN), a spring onion variety of southern Italy, is rich in antioxidant and bioactive molecules, yet its effects on hepatic steatosis remain unexplored. This study evaluated the hepatoprotective activity of a CN leaves extract in HepG2 cells exposed to oleic and palmitic acids (OA/PA) to induce non-alcoholic fatty liver steatosis, in both two- and three-dimensional culture systems. UHPLC-HRMS/MS analysis revealed a complex phytochemical profile comprising polar and apolar lipids, chlorophylls, carotenoids, and flavonoids. These metabolites, known to regulate oxidative stress, inflammation, and lipid metabolism, likely mediate CN’s protective effects against lipotoxicity. In HepG2 cells, CN markedly improved cell viability, attenuated lipid droplet accumulation, restored redox balance, alleviated endoplasmic reticulum (ER) stress, preserved mitochondrial membrane potential, reduced inflammation markers, and significantly modulated lipid-metabolism–related gene expression. Sirtuin 1 (SIRT1) expression and activity were enhanced by CN in the presence of OA/PA, and the pharmacological modulation confirmed its partial involvement in the protective effects. These preliminary findings demonstrate that CN counteracts free fatty acid-induced lipotoxicity through an integrated mechanism targeting lipid metabolism, oxidative and ER stress, mitochondrial function, and inflammation, with SIRT1 activation acting as a central mediator supporting its potential as a promising, sustainable nutraceutical for the prevention or adjunct management of non-alcoholic fatty liver disease.