The Regulatory Effects of Ferroptosis Inhibitors on Cardiomyocyte Apoptosis Via the PI3K/AKT/ROS Signaling Pathway

LONG Xiao; CHEN Ran; YANG Tao; HUANG Yuyan; TANG Changchun

doi:10.12102/j.issn.1672-1349.2025.15.009

Abstract：Objective:To investigate the effect of ferrostatin-1(Fer-1)on cardiomyocyte apoptosis in cardiovascular diseases through reactive oxygen(ROS)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway.Methods:Rat H9c2 cardiomyocytes were randomly assigned to the following experimental groups:the control group,the hypoxia/reoxygenation(H/R)group,the H/R+phosphate-buffered saline(PBS)group,the H/R+low-dose Fer-1 group,the H/R+medium-dose Fer-1 group,and the H/R+high-dose Fer-1 group.The H/R group,H/R+PBS group,H/R+low-dose Fer-1 group,H/R+medium-dose Fer-1 group,and H/R+high-dose Fer-1 group were subjected to H/R induction in H9c2 cells,and the levels of lactate dehydrogenase(LDH),oxidative stress,and intracellular ferrous iron(Fe2+)were determined.The levels of ferroptosis markers glutathione peroxidase 4(GPX4)and acyl-coa synthase 4(ACSL4)were detected by Western Blot.H/R-induced H9c2 cells were treated with Fer-1 at concentrations of 1,3,and 12 μmol/L,and cell viability was detected.The expression of PI3K/AKT signaling pathway was detected by real-time fluorescent quantitative polymerase chain reaction and Western Blot.Results:The results showed that H/R-induced H9c2 cell viability decreased with increase of oxidative stress and lactate dehydrogenase content,elevation of Fe2+and ACSL4 levels,and decrease of GPX4 levels.Fer-1 inhibits H/R-induced ferroptosis and oxidative stress in cardiomyocytes.The expression of PI3K/AKT signaling pathway decreased in H/R-induced H9c2 cells,while Fer-1 treatment increased its expression,and the differences were statistically significant(P＜0.05).Conclusion:Fer-1 attenuates H/R-induced iron death in cardiomyocytes by activating the PI3K/AKT signaling pathway.