Scoparone ameliorates metabolic dysfunction-associated steatotic liver disease by regulating the miR-3073a-3p/CAMKK2 axis

Scoparone (SCO) is a bioactive coumarin compound isolated from the traditional medicinal plant Artemisia dracunculus L. var. turkestanica Krasch. Previous studies have demonstrated that SCO ameliorates hepatic steatosis. However, the precise molecular mechanisms underlying its action, especially in metabolic dysfunction-associated steatotic liver disease (MASLD), remain incompletely elucidated. The study aimed to reveal the therapeutic effects and underlying mechanisms of SCO in treating MASLD. In this study, an in vivo MASLD model was constructed in mice through high-fat diet (HFD) induction, and an in vitro steatosis model was established by treating AML-12 hepatocytes with sodium oleate (NaOA). Hepatic histopathological characteristics, lipid deposition and biochemical parameters of all experimental groups were detected, differentially expressed genes and miRNAs were screened through RNA and miRNA sequencing, and the direct targeting effect of miR-3073a-3p on Camkk2 was verified using dual-luciferase reporter gene assay. Meanwhile, Western blot and qRT-PCR were applied to determine cellular functional indicators as well as the protein and mRNA expression levels of key signaling molecules. miR-3073a-3p directly bound to the 3′UTR of Camkk2 mRNA, suppressing CAMKK2 expression and inhibiting CAMKK2/AMPK signaling. Overexpression of miR-3073a-3p exacerbated hepatic steatosis, mitochondrial dysfunction, oxidative stress, bile acid imbalance and autophagy impairment, whereas miR-3073a-3p knockdown ameliorated MASLD phenotypes in a CAMKK2-dependent manner. SCO treatment downregulated miR-3073a-3p expression, restored CAMKK2 levels and activated protective pathways mediated by Nrf2, FXR and ULK1, thereby enhancing antioxidant defense, restoring bile acid homeostasis, promoting autophagy and alleviating liver injury. By modifying the miR-3073a-3p/CAMKK2 axis, SCO improved MASLD, indicating its potential therapeutic utility in the management of metabolic liver disorders. • Scoparone (SCO) alleviated ferroptosis, bile acid dysregulation, and autophagy impairment in metabolic dysfunction-associated steatotic liver disease (MASLD). • MiR-3073a-3p directly bound to Camkk2 mRNA and suppressed the CAMKK2/AMPK pathway. • SCO downregulated miR-3073a-3p expression, which restored CAMKK2 levels and activated AMPK signaling. • Targeting the miR-3073a-3p/CAMKK2 axis with SCO was a promising therapeutic strategy for MASLD.