Protective Effect Of Acetoacetate On Hydrogen Peroxide Induced PC12 Cells Apoptosis And Oxidative Stress

The aim of the present article is to study possible effects of acetoacetate, one member of the ketone bodies, on oxidative damage in neuron cells. The in vitro model of hydrogen peroxide-induced oxidative damage was used in this study to investigate whether the neuroprotective potentials of acetoacetate and the possible mechanisms. From the results we could found that, 0.1-8.0 mmol/L acetoacetate has no cytotoxicity on PC12 cell, suggesting that such concentrations of acetoacetat can be used as a safe dose for the following experiments. By MTT assay, lactate dehydrogenase leakage test and Annexin V/PI apoptosis analysis, we found that 150 μmol/L hydrogen peroxide caused significant oxidative injury and increased apoptosis rate in PC12 cells, however, 0.5-4.0 mmol/L acetoacetate pretreatment effectively inhibits cell damage and cell apoptosis caused by hydrogen peroxide. Hydrogen peroxide-induced cell damage is associated with oxidative stress and oxidative stress-mediated activation of apoptosis proteins including caspase-9 and caspase-3. By the analysis of intracellular reactive oxygen species(by DCFH-DA fluorescent probe), antioxidant enzymes SOD and CAT(by enzyme activity detection kits), apoptosis proteins caspase-9 and caspase-3(by Western blot), we found that 0.5-4.0 mmol/L acetoacetate pretreatment can effectively prevent oxidative stress and apoptosis protein activation caused by hydrogen peroxide.Considering the comprehensive results, it can be confirmed that 0.5-4.0 mmol/L acetoacetate is able to inhit cell death and LDH leakage, reduce the apoptosis rate, resist the increase of intracellular reactive oxygen species, restore the activities of antioxidant enzymes, suppress the activation of apoptosis proteins. In conclusion acetoacetate showed good protective effects on hydrogen peroxide-induced oxidative damage and apoptosis on PC12 neuron-like cells. This study provides the theory basis for understanding of the physiological effect of acetoacetate, and provides drug selection for the treatment of neurodegenerative diseases, ischemic, anoxia or reperfusion damage at the same time.