Linarin alleviates high-fat diet-induced hepatic steatosis by inhibiting PDE4D and activating the cAMP/PKA/CREB pathway.

Metabolic-associated fatty liver disease (MAFLD), a major consequence of obesity and metabolic dysfunction, lacks effective treatments. The cAMP/PKA signaling pathway regulates lipid metabolism, and inhibition of its upstream target PDE4D alleviates MAFLD progression. Linarin, a natural flavonoid glycoside with hepatoprotective properties, remains underexplored for MAFLD mechanisms. To investigate linarin's therapeutic effects on high-fat diet (HFD)-induced MAFLD and elucidate its mechanisms, focusing on PDE4D inhibition and cAMP/PKA/CREB pathway activation. C57BL/6J mice were fed a normal diet (CON), high fat diet (HFD), HFD + 50 mg/kg linarin, and HFD + 100 mg/kg linarin. Oleic/palmitic acid-stimulated HepG2, Mouse primary cells and AML12 cells were used. In vitro, 25 μM linarin reduced intracellular triglycerides (TG), elevated ATP production, decreased ROS and MDA, and upregulated GSH and CAT. In vivo, 50 and 100 mg/kg linarin suppressed weight gain, reduced hepatic fat deposition, and improved insulin sensitivity and liver function. Mechanistically, linarin inhibits PDE4D activity, activates the cAMP/PKA/CREB pathway, and upregulates GPX4 expression. Linarin alleviates MAFLD by targeting PDE4D to activate the cAMP/PKA/CREB pathway, improving lipid metabolism, mitochondrial function, and oxidative stress. This study highlights the potential of natural compounds for metabolic disease intervention, providing a foundation for clinical translation.