Background: Bempedoic acid is a cholesterol synthesis inhibitor used for the treatment of hyperlipidemia. It acts upstream of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase to inhibit cholesterol biosynthesis and increase LDL receptor expression, it activates Adenosine 5’-monophosphate (AMP)-activated protein kinase (AMPK) and inhibits ATP-citrate lyase (ACL), resulting in lower LDL cholesterol and atherosclerosis in pre-clinical models. AMPK is also an upstream activator of endothelial NO synthase (eNOS), which is phosphorylated on Ser1177 in endothelial cells by several stimuli including metformin, adiponectin and leptin. However, it remains unknown whether bempedoic acid promotes endothelial function through eNOS activation, and the molecular mechanisms involved in this pathway. Methods: We have characterized the effects of bempedoic acid in HUVEC cells (human umbilical vein endothelial cells) by evaluating the phosphorylation of AMPK at Thr172, AMPK substrate ACC at Ser79, and eNOS at Ser1177. The cells were exposed to increasing concentrations of bempedoic acid (10, 25, 50 and 100 μM) for 24 hours. We treated HUVECs with AICAR 2mM for 24 hours, to obtain a visual positive control, and we pre-treated the cells for 1 h with Compound C (40 μM) as a negative control. We will use small inhibitory RNA (siRNA) to evaluate if the down-regulation of AMPK regulatory subunit β1 or β2 impairs bempedoic acid-induced eNOS phosphorylation in HUVECs. Moreover, we will analyze eNOS activity, by assessing the intracellular formation of L-[14C]citrulline from L-[14C]arginine, and measure intracellular cGMP. Results: Our preliminary data show that bempedoic acid treatment significantly increased AMPK (Thr172) and ACC (Ser79) phosphorylation levels in a dose-dependent manner, with maximal effect occurring at 100μM (p<0.05). HUVECs exposed for 24 hours to bempedoic acid showed a dose-dependent increase of eNOS (Ser1177) phosphorylation (100μM, p<0.001). Conclusion: The results of this project support the hypothesis of that bempedoic acid exerts beneficial actions on the endothelial system and will help shed new light on the molecular mechanisms involved and lay the basis for future development of therapeutic and preventive strategies for reducing the burden of cardiovascular risk.