Objective Hepatic stellate cells (HSCs) are the main producers of fibrotic extracellular matrix (ECM) and play a critical role in the initiation, progression, and regression of hepatic fibrosis (HF). Dihydromyricetin (DMY) has hepatoprotective properties, but the mechanism of this protection is unclear. Our study examined the effects of DMY on the activation of HSCs triggered by ferric ammonium citrate (FAC) in HSC-T6 cells and explored the possible mechanisms of the hepatoprotective properties of DMY.Methods Cell viability was evaluated using MTT assay. The levels of ECM in the culture supernatant were examined usingenzyme linked immunosorbent assay. The iron deposition levels in HSC-T6 cells were assessed using Prussian Blue staining. The total iron and free iron levels in HSC-T6 cells were measured using colorimetric assay and calcein-AM assay, respectively. The ultrastructure of HSC-T6 cells was observed using transmission electron microscopy. The expression levels of ferritin heavy chain 1 (FTH1), α-smooth muscle actin (α-SMA), nuclear receptor coactivator 4 (NCOA4), microtubule-associated protein 1 light chain 3(LC3), and p62/SQSTM1 were measured using Western blotting.Results Compared with the FAC group, the DMY+FAC group had a significant reduction in the main components of ECM, total iron and free ironlevels, expression levels of α-SMA, NCOA4, and LC3-Ⅱ proteins, and the ratio of LC3-Ⅱ/LC3-Ⅰ; there was also a significant upregulation of the protein expression levels of FTH1 and p62. Furthermore, rapamycin partially blocked the effects of DMY, which inhibited the activation of HSCs induced by FAC.Conclusion DMY inhibits the activation of HSCs induced by iron overload, and the underlying mechanism may be involved in the inhibition of ferritinophagy.