Ferristatin II Inhibits Ferroptosis by Regulating the Bone Morphogenetic Protein 6/SMAD Signaling Pathway in HepG2 Cells

Abstract

The aim of this study was to investigate the protective effect of Ferristatin II (Fer II) on ferric ammonium citrate (FAC)-induced ferroptosis and explore its mechanism by performing experiments in vitro. The cell viability of Fer II in the treatment of FAC-induced ferroptosis was investigated by MTT, measuring the concentrations of Fe2+ and MDA and the activity of CSH-PX. We further measured the protein expression of hepcidin (Hepc), TfR1, BMP6, p-Smad1 and p-Smad5 using Western blotting. The gene expression level of Hepc was significantly increased and the protein expression levels of p-SMAD1 and p-SMAD5 were also significantly up-regulated after the coordinated intervention of Fer II and BMP. The results showed that cell viability was increased after treatment with Fer II. The concentrations of Fe2+ and MDA revealed that Fer II decreased hepatocyte ferroptosis induced by FAC. The Western blot results also showed that Fer II up-regulated the protein expression of Hepc and down-regulated protein expression of TfR1, BMP6, p-Smad1 and p-Smad5. Further results showed that Fer II and BMP6 synergistically promoted Hepc secretion and up-regulated the protein expression levels of Smad1 and p-Smad5. Fer II alleviated FAC-induced ferroptosis in HepG2 cells by regulating the BMP6/SMAD pathway, suggesting a new therapeutic approach for hepatocyte protection.