Artesunate alleviates liver fibrosis by regulating ferroptosis signaling pathway

Abstract

)-induced mouse liver fibrosis and elucidate whether artesunate could alleviate liver fibrosis by regulating ferritinophagy- mediated ferroptosis in hepatic stellate cells (HSCs). Firstly, our results demonstrated that artesunate treatment could induce activated HSC ferroptosis in fibrotic livers. Moreover, primary HSCs isolated from different animal groups were cultured to detect biomarkers of ferroptosis including iron, lipid peroxidation, glutathione (GSH) and prostaglandin endoperoxide synthase 2 (ptgs2) levels. The results revealed that artesunate remarkably promoted ferroptosis of activated HSCs. Furthermore, consistent with the experimental results in vivo, the data in vitro still indicated that artesunate treatment markedly induced ferroptosis in activated HSCs, which mainly embodied as declined cell vitality, increased cell death rate, accumulated iron, elevated lipid peroxides and reduced antioxidant capacity. Conversely, inhibition of ferroptosis by deferoxamine (DFO) completely abolished artesunate-induced anti-fibrosis effect. Surprisingly, artesunate also evidently triggered ferritinophagy accompanied by up-regulation of LC3 (microtubule-associated protein light chain 3), Atg3, Atg5, Atg6/beclin1, Atg12 (autophagy related genes) and down-regulation of p62, FTH1 (ferritin heavy chain), NCOA4 (nuclear receptor co-activator 4) in activated HSCs. Nevertheless, depletion of ferritinophagy by specific inhibitor lysosomal lumen alkalizer-chloroquine (CQ) inhibited artesunate-induced ferroptosis and anti-fibrosis function. These results suggested that ferritinophagy-mediated HSC ferroptosis was responsible for artesunate-induced anti-fibrosis efficacy, which provided new clues for further pharmacological study of artesunate.