Alisol B 23-acetate alleviates high-fat diet-induced insulin resistance by activating the SIRT1/FOXO1 axis and PI3K/AKT pathway.

Metabolic dysfunction-associated steatotic liver disease (MASLD), the most frequent chronic liver disease, is strongly linked to insulin resistance and currently has no effective treatments available. Alisol B 23-acetate (AB23A), a key bioactive component from Alismatis Rhizoma, shows promising lipid-lowering properties, but its impact and mechanisms on MASLD are not fully elucidated. An in vivo model was established via high-fat diet (HFD)-fed mice, and an in vitro model was generated via oleic acid (OA)-induced AML-12 hepatocytes. The efficacy of AB23A was assessed by measuring fasting blood glucose (FBG)/insulin levels, lipid profiles, and hepatic lipid content. Network pharmacology analysis predicted potential targets, molecular docking, molecular dynamics simulations, biolayer interferometry (BLI), and a cellular thermal shift assay (CETSA) were used to validate the binding of AB23A and SIRT1. Protein expression was analyzed via Western blotting, immunohistochemistry, and immunofluorescence staining. AB23A significantly reduced FBG, fasting insulin levels, HOMA-IR, and hepatic lipid accumulation in HFD-fed mice, while simultaneously improving liver function. Network pharmacology analysis combined with experimental validation demonstrated that AB23A directly binds to and activates SIRT1. In vitro, AB23A alleviated OA-induced hepatic steatosis and insulin resistance. Importantly, the SIRT1 inhibitor EX527 abolished the beneficial effects of AB23A, including the upregulation of the p-AMPK, p-ACC, CPT1α, and p-FOXO1, as well as the activation of the PI3K/AKT pathways. This study identified SIRT1 as a direct target of AB23A. AB23A ameliorates hepatic steatosis and insulin resistance by targeting SIRT1, promoting fatty acid oxidation, modulating downstream FOXO1 activity, and coactivating the PI3K/AKT signaling pathway. These findings highlight AB23A as a promising therapeutic candidate for MASLD.