Dihydromyricetin attenuates cholestatic liver injury by suppressing JAK2/STAT3-associated NF-κB inflammatory and apoptotic signaling

BACKGROUND: Cholestatic liver injury (CLI) is mainly driven by hepatic bile acid accumulation, which promotes hepatocyte injury, inflammation, oxidative stress, and apoptosis. Dihydromyricetin (DHM), a bioactive flavonoid from Ampelopsis grossedentata, has shown hepatoprotective potential, but its role and mechanism in CLI remain unclear. METHODS: Network pharmacology, public transcriptomic analysis, and molecular docking were used to predict candidate targets of DHM in CLI. Taurocholic acid (TCA) treated HepaRG cells and α-naphthylisothiocyanate (ANIT)-treated mice were used as in vitro and in vivo models, respectively, with ursodeoxycholic acid (UDCA) included as a positive control. Hepatic injury, inflammation, oxidative stress, apoptosis, JAK2/STAT3-associated NF-κB signaling, bile acid-related molecules, and ductular reaction markers were assessed. AG490-mediated pathway inhibition and IL-6-mediated STAT3 reactivation were performed to assess the functional involvement of STAT3 signaling. RESULTS: DHM improved cell viability and reduced TCA-induced ALT, AST, LDH, ALP, and inflammatory cytokine levels. Network pharmacology identified STAT3, AKT1, SRC, ESR1, and MMP9 as hub targets, while public transcriptomic and enrichment analyses highlighted inflammation-, apoptosis-, NF-κB-, and JAK/STAT-related pathways. DHM attenuated TCA-induced apoptosis and suppressed JAK2/STAT3-associated NF-κB activation, including reduced p65 nuclear translocation. DHM also regulated bile acid homeostasis-related molecules in vitro. In ANIT-treated mice, DHM improved liver histology, decreased ALT, AST, ALP, TBA, inflammatory cytokines, and oxidative stress, and suppressed hepatic apoptosis and JAK2/STAT3-NF-κB signaling. AG490 phenocopied the protective effects of DHM, whereas IL-6-mediated STAT3 reactivation partially weakened DHM-mediated protection. CONCLUSION: DHM alleviates CLI by suppressing inflammation, oxidative stress, and hepatocyte apoptosis, at least partly through inhibition of the JAK2/STAT3-NF-κB signaling axis.