Phytoestrogen alpha-zearalanol attenuate endoplasmic reticulum stress to against cultured rat hippocampal neurons apoptotic death induced by amyloid beta25-35.

Objective: Our previous studies demonstrated both phytoestrogen α-zearalanol (α-ZAL) and estrogen is effective decrease Alzheimer's disease (AD)-like apoptotic neuron death, but α-ZAL showed significantly less side-effect on breast and endometrial tissue compared to estrogen, it suggested that α-ZAL can be used as a potential substitute for estrogen. However, the molecular mechanism by which α-ZAL prevents neuron damage remains unclear. Growing evidence suggests that endoplasmic reticulum (ER) stress plays an important role in the process of cell apoptosis in AD; in addition, our published data indicated that α-ZAL possessed the potential ability to stabilize ER function. We therefore hypothesized that ER-stress mechanism maybe involved in the antiapoptotic effect of α-ZAL in this study.

Methods: Primary rat hippocampal neurons have been cultured and subsequently followed exposed to β-peptide fragment 25-35(Aβ25-35) with or without α-ZAL pre-treatment, and then western blot and flow cytometry techniques has been used to evaluate the intracellular calcium balance, ER stress and apoptotic cell death.

Results: The results showed that Aβ25-35 treatment for 24h induced dramatic neuronal apoptosis, accompanied by an increase in calpain2 expression, a marker of intracellular calcium overload. On the other hand, ER stress sensitive hallmarks, glucose-regulated protein 78 (GRP78), double-stranded RNA-dependent protein kinase (PKR)-like ER-resident kinase (PERK) and C/EBP homologous protein-10 (CHOP10) expressions were up-regulated after Aβ25-35 administration. Importantly, α-ZAL pre-treatment effectively attenuated above changes.

Conclusion: These results demonstrated that α-ZAL protects cells against AD-like apoptosis and the effects at least partially by attenuating severely ER stress.