Nuciferine Exerts Anti‐Inflammatory Effects in Mice With Non‐Alcoholic Steatohepatitis by Regulating the miR‐23a‐3p‐SIRT1‐NF‐κB Pathway and Akkermansia muciniphila‐Extracellular Vesicles

ABSTRACT Nuciferine, a lotus leaf extract with low bioavailability, shows beneficial effects on hepatic metabolism and gut microbiota, but its anti‐inflammatory mechanisms in non‐alcoholic steatohepatitis (NASH) are unclear. This study aimed to clarify how nuciferine mitigates hepatic inflammation in NASH by exploring its interactions with immune pathways and gut microbiota. Initially, a NASH mouse model was induced using a methionine‐ and choline‐deficient diet, with nuciferine administered orally. Furthermore, liver damage was assessed, and hepatic M1 (CD11B + pro‐inflammatory) and M2 (CD163 + anti‐inflammatory) macrophages were quantified. Molecular assays measured SIRT1 gene expression, while miRNA sequencing and dual‐luciferase assays explored its role in the SIRT1/NF‐κB pathway. Additionally, gut microbiota were analyzed via 16S rRNA sequencing, and fluorescently labeled Akkermansia muciniphila ‐derived extracellular vesicles ( Akk ‐EVs) were tracked in vivo and in vitro. Treatment with nuciferine reduced liver injury, decreasing pro‐inflammatory M1 macrophages and increasing anti‐inflammatory M2 macrophages. Meanwhile, it upregulated hepatic SIRT1, suppressing miR‐23a‐3p to inhibit the NF‐κB pathway and promote M1‐to‐M2 polarization. Gut microbiota analysis showed nuciferine enriched Akkermansia muciniphila , and fluorescent imaging confirmed Akk ‐EVs entered liver tissues and macrophages, exerting direct anti‐inflammatory effects. In conclusion, nuciferine protects against NASH through dual mechanisms: modulating the SIRT1/NF‐κB pathway to reduce hepatic inflammation and enhancing Akk‐ EVs. These findings highlight its therapeutic potential for NASH, linking host immune responses with gut microbiota interactions.