Bioactive polymethoxylated flavones from Clerodendranthus spicatus alleviate high-fat diet-induced lipid dysfunction...

The high-fat diet (HFD) pattern is currently a major cause of chronic metabolic diseases worldwide, especially metabolic dysfunction-associated steatohepatitis (MASH). Clerodendranthus spicatus (Thunb.) C. Y. Wu (CST) is an edible-medicinal plant widely used for health promotion, however its bioactive constituents and mechanisms against HFD-induced metabolic disorders have not been convincingly demonstrated. In this study, the ethyl acetate fraction of CST extract (EAF_CST), rich in polymethoxylated flavones (PMFs), showed better lipid-lowering and anti-inflammatory activities than raw extract. Subsequent in vivo experiments confirmed that EAF_CST markedly alleviated HFD-induced obesity (body weight reduced by 17% in the high-dose group, p < 0.001 vs. HFD), insulin resistance, dyslipidemia (serum TG lowered by 40% in the high-dose group, p < 0.001 vs. HFD), hepatic steatosis and inflammation in rats. Further integrative transcriptomic and network analysis revealed that tetramethylscutellarein, trimethylapigenin and eupatorin-5-methylether strongly interacted with lipid/inflammation-related hub targets and pathways (notably the PPAR signaling pathway), leading to their designation as the core bioactive constituents. Geographical commonality and efficacy validation further reinforced this designation. Mechanistically, these PMFs directly bound to PPARα, thereby enabling EAF_CST to promote fatty acid oxidation, and suppress NF-κB-mediated inflammatory response. Finally, the therapeutic effects of EAF_CST were further verified to depend on PPARα. This study establishes a clear constituent-target-pathway axis for CST's metabolic benefits, and the first evidence that these benefits are mediated by the core PMFs targeting PPARα. These findings also provide a scientific basis for developing CST into functional foods/nutraceuticals for combating HFD-related disorders (e.g., MASH).