Apolipoprotein E4 exacerbates ethanol-induced neurotoxicity through augmentation of oxidative stress and apoptosis in N2a-APP cells

Neuronal loss is a prominent phenomenon in Alzheimer's disease (AD) and alcohol-induced brain damage. Alcohol abuse is associated with an increased risk of AD, regardless of the type of alcoholic beverage. Furthermore, the detrimental effect of excessive alcohol consumption on the risk of AD is exacerbated among people carrying apolipoprotein E (APOE) epsilon 4 allele, the major genetic risk factor for AD. However, how APOE epsilon 4 and alcohol abuse synergistically enhance the possibility of AD is unclear. Here we show that in N2a cells stably expressing human APP695 (N2a-APP), high-concentration ethanol-induced neurotoxicity was significantly augmented in the presence of apoE4 protein, compared with apoE3 protein. Early and late apoptotic cells were apparently more in cells treated with the combination of apoE4 and ethanol, compared with that of apoE3 and ethanol. Inhibition of apoptosis using a pan-caspase inhibitor z-vad resulted in abolishment of the apoE isoform-specific effect on high-concentration ethanol-induced neurotoxicity. Moreover, compared with apoE3, apoE4 augmented ethanol-induced cellular oxidative stress, and pre-incubation with a reactive oxygen species (ROS) scavenger NAC abrogated the specific effect of apoE4 on ethanol-induced neurotoxicity. Taken together, our results for the first time demonstrate that apoE4 and high-concentration ethanol synergistically enhance neurotoxicity through elevating cellular oxidative stress and increasing neuronal apoptosis, and support the notion that avoiding excessive alcohol consumption can help to prevent AD especially in APOE epsilon 4 carriers.