Supplementary Material for: Actein, a 9,19-cyclobutane triterpenoid from Cimicifuga spp., ameliorates atherosclerosis via ABCG1 and LXR upregulation

Introduction. Considering the global burden of atherosclerosis-related cardiovascular mortality, this study investigates the anti-atherosclerotic potential of Actein, a 9,19-cyclobutane triterpenoid isolated from Cimicifuga spp. Methods. The study employed both in vitro and in vivo experiments. In vitro, the effect of Actein on oxLDL-induced macrophage foam cells was examined. In vivo, ApoE-deficient mice were fed a high-fat diet to induce atherosclerosis and were then treated with Actein (10 mg/kg) or Atorvastatin for 8 weeks. Results. Actein significantly reduced lipid accumulation in oxLDL-induced foam cells. In the in vivo model, Actein treatment significantly lowered serum levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol, while increasing high-density lipoprotein cholesterol. Furthermore, Actein diminished the levels of inflammatory markers (IL-6, IL-1β, TNF-α, and MMP-9) and upregulated the expression of ABCG1 and LXR proteins in liver tissue. The efficacy of Actein was comparable to, and in some aspects superior to, that of Atorvastatin. Conclusion. These findings establish Actein as a promising natural compound for the treatment of atherosclerosis, warranting further investigation into its therapeutic potential.