Ginsenoside Rh4 prevents endothelial dysfunction as a novel AMPK activator

Background and Purpose The AMP‐activated protein kinase (AMPK) signalling pathway is a desirable target for various cardiovascular diseases (CVD), while the involvement of AMPK‐mediated specific downstream pathways and effective interventions in hyperlipidaemia‐induced endothelial dysfunction remain largely unknown. Herein, we aim to identify an effective AMPK activator and to explore its efficacy and mechanism against endothelial dysfunction. Experimental Approach Molecular docking technique was adopted to screen for the potent AMPK activator among 11 most common rare ginsenosides. In vivo, poloxamer 407 (P407) was used to induce acute hyperlipidaemia in C57BL/6J mice. In vitro, palmitic acid (PA) was used to induce lipid toxicity in HAEC cells. Key Results We discovered the strongest binding of ginsenoside Rh4 to AMPKα1 and confirmed the action of Rh4 on AMPK activation. Rh4 effectively attenuated hyperlipidaemia‐related endothelial injury and oxidative stress both in vivo and in vitro and restored cell viability, mitochondrial membrane potential and mitochondrial oxygen consumption rate in HAEC cells. Mechanistically, Rh4 bound to AMPKα1 and simultaneously up‐regulated AKT/eNOS‐mediated NO release, promoted PGC‐1α‐mediated mitochondrial biogenesis and inhibited P38 MAPK/NFκB‐mediated inflammatory responses in both P407‐treated mice and PA‐treated HAEC cells. The AMPK inhibitor Compound C treatment completely abrogated the regulation of Rh4 on the above pathways and weakened the lowering effect of Rh4 on endothelial impairment markers, suggesting that the beneficial effects of Rh4 are AMPK dependent. Conclusion and Implications Rh4 may serve as a novel AMPK activator to protect against hyperlipidaemia‐induced endothelial dysfunction, providing new insights into the prevention and treatment of endothelial injury‐associated CVD.