Magnoflorine Ameliorates MASH in Mice Fed a High‐Fat and High‐Fructose Diet by Promoting AMPK ‐Mediated Lipophagy and Fatty Acid Oxidation

Emerging as a significant clinical burden, metabolic dysfunction-associated steatohepatitis (MASH) continues to challenge global healthcare systems due to limited effective interventions. Magnoflorine, a naturally occurring quaternary aporphine alkaloid abundant in dietary botanicals, has demonstrated promising hypolipidemic and anti-inflammatory properties in preliminary investigations. This investigation systematically evaluated both the therapeutic potential and mechanistic pathways underlying the capacity of magnoflorine to ameliorate MASH-associated hepatic pathologies. Male C57BL/6 mice received 12-week high-fat and high-fructose diet (HFFD) induction to establish MASH models, with graded magnoflorine dosing. Through integrated histopathology (H&E/ORO/Masson), transcriptomics, and targeted interventions at both cellular and animal levels, we elucidated the therapeutic mechanisms of magnoflorine. Results showed that magnoflorine effectively alleviated hepatic steatosis, inflammation, and fibrosis in HFFD-fed mice. Mechanistically, RNA-seq revealed that the 5'-monophosphate AMP-activated protein kinase (AMPK) activation was the central pathway mediating the effects of magnoflorine to impede MASH progression. Furthermore, AMPKα activation induced by magnoflorine stimulated hepatic lipophagy in HFFD-fed mice, which further affected lipid β-oxidation and inflammation. The findings were consistent in HepG2 cells incubated with palmitic acid. In vitro, silencing AMPKα reversed the stimulatory effect of magnoflorine on lipophagy in HepG2 cells. In vivo, liver-specific AMPKα silencing in mice blocked the enhancement of hepatic lipophagy and the improvement of MASH caused by magnoflorine. In brief, this study indicates that magnoflorine improves MASH in mice by activating hepatic lipophagy via AMPKα-dependent effects. This discovery provides a novel potential therapeutic alternative for MASH.