Mechanism of AKT/AMPK/Nrf2 Regulated by Loganin to Improve Ferroptosis Induced by Oxygen Glucose Deprivation/Reoxygenation
Objective:To investigate the mechanism of loganin improves the ferroptosis induced by oxygen glucose deprivation/reoxygenation(OGD/R)by regulating protein kinase B(AKT)/adenylate activated protein kinase(AMPK)/nuclear factor E2 associated factor 2(Nrf2)pathway.Methods:The neurons were divided into control group,OGD/R group,OGD/R+L-loganin group,OGD/R+M-loganin group,OGD/R+ H-loganin group,OGD/R+ H-loganin+ ML385 group.The morphology of neuron mitochondria was observed by transmission electron microscope.Iron content,glutathione peroxidase 4(GPX4)activity,4-hydroxynonenal(4-HNE),superoxide dismutase(SOD),malondialdehyde(MDA),reduced glutathione(GSH)/oxidized glutathione(GSSG),reduced Coenzyme Ⅱ(NADPH)/Codehydrogenase Ⅱ(NADP+),and Lactate dehydrogenase(LDH)content were determined.Intracellular and lipid reactive oxygen species(ROS)levels were detected by CM-H2DCFDA and C11-BODIPY581/591,respectively.MTT assay was used to detect cell activity.B-cell lymphoma 2(Bcl-2),Bcl-associated X protein(Bax),cleaved Caspase-3,phosphorylated protein kinase B(p-AKT)/AKT,phosphorylated adenylate activated protein kinase(p-AMPK)/AMPK,Nrf2 protein expression were detected by Western Blot.Results:In OGD/R group,mitochondrial fragmentation occurred,ridge decreased,and mitochondrial membrane density increased.Compared with the control group,the relative activities of GSH/GSSG,NADPH/NADP+,SOD,GPX4,and the levels of Bcl-2,p-AKT/AKT,p-AMPK/AMPK and Nrf2 in OGD/R group decreased(P<0.05),Fe2+content,intracellular ROS levels,lipid ROS levels,4-HNE,MDA levels,LDH release,Bax,and cleaved Caspase-3 levels increased(P<0.05).After treatment with loganin,the mitochondrial ridge in neuronal mitochondria became more complete and the fragmentation disappeared.The relative activity of GSH/GSSG,NADPH/NADP+,SOD,GPX4,Bcl-2 level,p-AKT/AKT,p-AMPK/AMPK,and Nrf2 in OGD/R+L-loganin group,OGD/R+M-loganin group and OGD/R+H-loganin group were higher than those in OGD/R group,Fe2+ content,intracellular ROS levels,lipid ROS levels,4-HNE,MDA levels,LDH release,Bax,and cleaved Caspase-3 levels were lower than those in OGD/R group(P<0.05).With the increase of the dose of loganin,the improvement effects were more significant.The above indexes of OGD/R+H-loganin+ML385 group were consistent with the trend of OGD/R group.Conclusion:loganin may ameliorate OGD/R-induced neuronal ferroptosis by regulating the AKT/AMPK/Nrf2 pathway.
oxyglucose deprivation/reoxygenationferroptosisloganinprotein kinase B/adenylate-activated protein kinase/nuclear factor-E2-related factor 2 pathwayneurons
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