Daphnetin Alleviates Liver Fibrosis by Inducing Ferritinophagy‐Mediated Ferroptosis in Activated Hepatic Stellate Cells

Liver fibrosis is an increasingly serious global health issue, and an effective treatment strategy is to inhibit the activation of hepatic stellate cells (HSCs). The role of daphnetin (Daph), which is a natural coumarin derivative isolated from plants of the genus Daphne, in the treatment of hepatic fibrosis is still unclear. Mice were treated with carbon tetrachloride (CCl₄), and the inhibitory effect of Daph on hepatic fibrosis was evaluated. In vitro, the effect of Daph on the activated human HSCs line LX-2 was studied. In vivo, Daph alleviated collagen accumulation and reduced the expression of fibrotic genes. However, these changes were reversed with ferrostatin-1 (Fer-1). In vitro, Daph induced ferroptosis in LX-2 cells, which was characterized by increased lipid peroxidation and iron accumulation. However, deferoxamine (DFO) and Fer-1 partially abrogated the antifibrotic effect of Daph. Mechanically, Daph exerted an antifibrotic effect by ubiquitinating glutathione peroxidase 4 (GPX4) and stimulating ferritinophagy-mediated ferroptosis in HSCs. These results indicated that Daph promoted the ubiquitination of GPX4 and ferritinophagy-mediated ferroptosis in HSCs, which could provide new clues for further pharmacological research on the antifibrotic effect of Daph.