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 PC12 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.