Deferoxamine Alleviates HT22 Cell Damage Induced by Hypoxia/Reoxygenation Injury Through Regulating Ferritinophagy
Objective To investigate the role of deferoxamine(DFO)in alleviating hypoxia/reoxygenation(HR)injury in mouse hippocampal neuronal cells(HT22)by regulating ferritinophagy mediated by nuclear receptor coactivator 4(NCOA4).Methods HT22 cell lines was randomly divided into control group using a random number table(Ctrl group),hypoxia/reoxygenation group(HR group),hypoxia/reoxygenation+DFO group(HR+DFO group).The HR group was cultured under sugar-free hypoxia for 6hours,followed by sugar-reoxygenation for 24hours to establish the neuronal HR model.The DFO group received a pre-treatment of 150μmol/L DFO for 24hours before HR.Immunofluorescence was used to detect live-dead cell staining and reactive oxygen species(ROS)levels.Enzyme-linked immunosorbent assay(ELISA)was used to detect the superoxide dismutase(SOD),malondialdehyde(MDA)and fer-rous ion levels.Autophagosomes were observed by electron microscopy.The expression levels of NCOA4,LC3B,Beclin-1 and ferritin was detectd by Western blot.Results Compared with the Ctrl group,the cell survival rate and SOD levels were decreased in HR group,MDA,ROS,NCOA4,LC3B,Beclin-1,autophagosomes,ferritin and ferrous ion content were increased(P＜0.05).Compared with the HR group,the cell survival rate and SOD levels were increased in HR+DFO group,MDA,ROS,NCOA4,LC3B,Beclin-1,auto-phagosomes,ferritin and ferrous ion content were decreased(P＜0.05).Conclusion DFO downregulates oxidative stress levels and fer-rous ion content by decreasing the expression of NCOA4,effectively alleviating HT22 cell damage caused by HR injury.
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