Isomucronulatol attenuates liver aging by inhibiting GSK-3β acetylation and promoting β-catenin accumulation.

Liver aging is a critical contributor to metabolic dysregulation and the development of various liver diseases. Currently, there is no approved therapy specifically targeting liver aging. The potential of medicinal and edible herbs in preventing and treating liver aging has attracted increasing attention. Accordingly, we constructed a self-built compound library containing 87 bioactive constituents isolated from these herbs. This study aimed to identify the potential compound capable of alleviating hepatic senescence from the constructed library and to elucidate the underlying mechanism. Isomucronulatol (Iso), a bioactive isoflavone isolated from Astragalus membranaceus, exhibited potential anti-hepatic senescence activity in a p21 promoter-driven luciferase reporter assay. Iso exhibited significant anti-hepatic senescence activity in H₂O₂-stimulated and doxorubicin (DOX)-stimulated HepG2 cells, as well as in mitomycin C (MC)-exposed AML12 cells, as evidenced by reduced SA-β-Gal positivity, downregulated p21 and p16 expression, and decreased secretion of SASP factors (IL-6, IL-1β, CCL2). In a D-galactose (D-gal)-induced mouse aging model, Iso treatment attenuated liver histological damage, lowered serum ALT and AST levels, and suppressed senescence markers in liver tissues. Transcriptomic and network pharmacology analyses revealed that Iso could activate Wnt/β-catenin signaling pathway to delay hepatic senescence. Mechanistically, Iso could facilitate nuclear accumulation of β-catenin by inhibiting GSK-3β acetylation, thereby restoring Wnt signaling. Collectively, these findings highlight Iso as a potential therapeutic agent against hepatic senescence.