5-Methoxyflavone alleviates high-fat diet-induced atherosclerosis in ApoE^-/- mice through modulation of ACADM

Atherosclerosis (AS) is a chronic vascular pathology and a leading cause of global cardiovascular morbidity and mortality, with dysregulated lipid metabolism acting as a primary driver of its progression. 5-Methoxyflavone (5-MF), a natural flavonoid, exhibits known lipid-modulating and antioxidant properties; however, its specific atheroprotective potential remains elusive. This study investigated the therapeutic efficacy and underlying metabolic mechanisms of 5-MF against AS. We employed a high-fat diet (HFD)-fed ApoE^-/- mouse model alongside a cholesterol-induced vascular smooth muscle cell (VSMC) injury model. Downstream molecular pathways were delineated using integrated bioinformatics and molecular assays. In vivo, 5-MF supplementation significantly ameliorated systemic glucolipid metabolism, reduced serum lipid profiles, and attenuated aortic lipid deposition and atherosclerotic plaque formation. In vitro, 5-MF suppressed cholesterol-induced lipid accumulation, oxidative stress, migration, and proliferation in VSMCs. Mechanistically, 5-MF promoted fatty acid oxidation (FAO) by upregulating medium-chain acyl-CoA dehydrogenase (ACADM) expression. Notably, ACADM overexpression diminished intracellular total cholesterol and triglyceride levels, whereas its knockdown exacerbated lipid deposition. In conclusion, these findings demonstrate that 5-MF confers anti-atherosclerotic effects by enhancing ACADM-mediated FAO, underscoring its promise as a novel nutritional intervention for AS prevention.