Doxofylline Attenuates Hydrogen Peroxide-induced Oxidative Damage in PC12 Cells by Activating AMPK Pathway
OBJECTIVE To investigate the protective effect of doxofylline against hydrogen peroxide(H2O2)-induced cytotoxicity in PC 12 and its underlying mechanism.METHODS The cell oxidative damage model was established by H2O2.MTT assay was used to detect the cell viability;Tunel staining,flow cytometry and Caspase-3 activation kit were used to detect cell apoptosis;Lactate dehydrogenase kit was used to detect cell membrane damage;Mitochondrial membrane potential was detected by JC-1 staining and the intercellular ROS level was detected by DCFH-DA staining;Western blotting was used to detect changes in signaling pathway protein levels.RESULTS Doxofylline concentration-dependently improved H2O2-induced abnormal levels of reactive oxygen species in PC12 cells,restored mitochondrial membrane potential,reduced Caspase-3 activation,and finally reduced PC12 cell apoptosis.Doxofylline also stimulated phosphorylation of AMPK in PC12 cells,and its protective effect was reversed by AMPK pathway inhibitors.CONCLUSION Doxofylline improves H2O2-induced PC12 cell injury by activating AMPK signaling pathway.
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