Ferroptosis Induced by H2O2 in L-O2 Cells Through Inhibition of the ERK Pathway
Objective To investigate the role and potential mechanism of hydrogen peroxide(H2O2)inducing ferroptosis in human liver parenchymal cells(L-O2 cells).Methods Different concentration gradients of H2O2 were used to stimulate L-O2 cells,and cell survival was detected using the CCK-8 assay after 12 and 24 hours of treatment to determine the opti-mal H2O2 induction conditions.Treatment groups included a blank control group,H2O2 group,Erastin group,and H2O2+Fer-1 group.It was determined that H2O2 could induce ferroptosis in L-O2 cells,and to further clarify the specific mecha-nism,the cells were divided into the blank control group,the H2O2 group,and the H2O2+ERK agonists of different con-centrations as well as ERK inhibitors.The kit was used to detect the cell death,ROS,and MMP in the treatment groups,and the mRNA expressions of ferroptosis markers PTGS2 and ACSL4 were detected using RT-qPCR.Results After stimu-lation with 800 μmol/L H2O2 for 24 hours,L-O2 cell death could be induced.Compared with the blank control group,the cells in the H2O2 and Erastin groups were significantly damaged,with elevated intracellular ROS,significantly reduced MMP,and increased mRNA expressions of PTGS2 and ACSL4.Compared with the H2O2 group,the cells in the H2O2+Fer-1 group had attenuated damage,reduced intracellular ROS,restored MMP,and down-regulated mRNA expressions of PTGS2 and ACSL4.Similarly,compared with the H2O2 group,cell damage was weakened in the H2O2+ERK agonist group,with reduced ROS,restored MMP,and lowered mRNA expressions of PTGS2 and ACSL4.However,after the addi-tion of the ERK inhibitor,opposite results were observed.Conclusion Treatment with 800 μmol/L H2O2 for 24 hours sig-nificantly induced ferroptosis in L-O2 cells,and this process may be mainly through the inhibition of the ERK pathway.
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