The neuroprotective effects of phosphocreatine on Amyloid Beta 25-35-induced differentiated neuronal cell death through inhibition of AKT /GSK-3β /Tau/APP /CDK5 pathways in vivo and vitro

Abstract

Alzheimer (AD) is a degenerative disease that can lead memory loss and behavioral dysfunction. Aβ protein and phosphorylation of Tau protein are related to the onset of AD. However, at present, its treatment and drugs are limited. The purpose of our study is to evaluate whether phosphocreatine (PCr) could protect neuronal injury induced by Aβ protein in vivo and in vitro through AKT/GSK-3β/Tau/APP/CDK5 pathways. Differentiated PC-12 cells were cultured with Aβ25-35 for 24 h, while the mice were injected with D-Galactose for eight weeks, both of them were pretreated with PCr for 2 h. The results showed PCr could obviously induce cells and hippocampus apoptosis using DAPI and TUNEL. PCr decreased the levels of intercellular reactive oxygen species (ROS) and malondialdehyde (MDA), and increased the activities of superoxide dismutase (SOD). Besides, the apoptosis pathway was detected using Western blot, showing that PCr could significantly reduce caspase-3, caspase-9, Bcl-2/Bax expression in vivo and in vitro. At the same time, PCr could decreased Ca²⁺ and apoptosis by Flow Cytometry in PC-12 cells. We observed that the morphological alteration of hippocampus injury was mitigated with the pretreatment of PCr. Furthermore, PCr pretreatment could decrease Aβ25-35-induced PC-12 cells apoptosis with APP cDNA transfection, which up-regulated AKT/GSK-3β/CDK5 pathways and induced Tau phosphorylation. In summary, PCr could reduce Aβ25-35 toxicity to protect neuronal cells via AKT/GSK-3β/CDK5 pathways.

Keywords: Alzheimer amyloid Beta 25–35 phosphocreatine tau.