Investigation of Europetin-3-O-Rhamnoside as a novel natural insulin-sensitizing compound in 3T3-L1 Adipocytes via regulation of GLUT-4 and adiponectin expression
Abstract
Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder characterized by impaired insulin signaling, reduced glucose uptake, oxidative stress, and adipocyte dysfunction. The present study investigated the antidiabetic potential of Europetin-3-O-rhamnoside as a novel natural insulin-sensitizing compound in high glucose-induced insulin-resistant 3T3-L1 adipocytes through modulation of GLUT4 and adiponectin-associated signaling pathways. Insulin resistance was induced in differentiated 3T3-L1 adipocytes using high-glucose conditions, followed by treatment with Europetin-3-O-rhamnoside at different concentrations. Cell viability was evaluated using the MTT assay, while gene expression analysis of insulin receptor (IR), insulin receptor substrate (IRS), phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and glucose transporter-4 (GLUT4) was performed using quantitative real-time PCR. Oxidative stress and antioxidant biomarkers including superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and hydrogen peroxide (H₂O₂) were also assessed. High-glucose exposure significantly reduced IR, IRS, PI3K, AKT, and GLUT4 expression levels while increasing oxidative stress, confirming the establishment of insulin resistance in adipocytes. Treatment with Europetin-3-O-rhamnoside significantly restored insulin-signaling gene expression in a dose-dependent manner, with the 20 µg/mL group exhibiting effects comparable to metformin-treated cells. Furthermore, Europetin-3-O-rhamnoside enhanced antioxidant defense systems by increasing SOD, CAT, and GSH levels while reducing H₂O₂ accumulation. The findings suggest that Europetin-3-O-rhamnoside ameliorates insulin resistance through restoration of the PI3K/AKT/GLUT4 signaling pathway and attenuation of oxidative stress-mediated cellular dysfunction. Overall, the study demonstrates that Europetin-3-O-rhamnoside possesses promising insulin-sensitizing and antioxidant properties and may serve as a potential natural therapeutic candidate for the management of insulin resistance and type 2 diabetes mellitus.