The Protective Effects Butylphthalide On Apoptosis Of PC12 Cells Induced By β-amyloid Peptide

ObjectiveAlzheimer's disease is a progressive neurodegenerative disorder characterized by the deposit of amyloid fibrils in the brain that result from the self-aggregative polymerization of the Aβ.However, significant methods for the treatment of the promotion and progression of AD are unavailable to date. Recent studies suggested that the redox imbalance and the accumulation of Aβpeptide occurring in the brain of AD patients lead to oxidatively-induced apoptotic cell death. Butylphthalide (NBP) is a potentially beneficial and promising drug for the treatment of ischemic stroke with multiple actions that affect different pathophysiological processes.Therefore, we show the effects of NBP on Aβ25-35-induced cytotoxicity in PC 12 cells, observed the apoptosis and mitochondrial changes of PC 12 cell, to explore the cell protective effect and possible mechanisms of NBP.MethodsCultured Cells PC 12 were divided into 3 groups:normal group, model group, NBP group.PC 12 cells were treated with different concentrations of NBP.PC12 cells were routinely cultivated and treated by Aβ25-35 (final concentration,10μmol/L) 2 hours before the addition of NBP. Forty-eight hours later, cells were examined viability and apoptosis by MTT method and HE,PI method, respectively. MAD and SOD activity detected by spectrophotometry to observe the effects of oxidative stress.The ultrastructural changes of neuronal mitochondria were viewed under transmission electron microscope. The expression of Bcl-2 and cytochrome C (Cyt-C) were detected by western blot.ResultsIt was revealed by MTT assay that various concentration of NBP could significantly prevent the cell viability induced by Aβ25-35 (P< 0.05). Pretreatment with lOμmol/LNBP could significantly inhibit the viability decreasing induced by Aβ25-35 (P<0.05).HE and PI show that the apoptosis rate of the NBP treated group is significantly lower than the model group.The activity of SOD in the NBP treated group was obviously higher than that in the model group, while MAD activity had the the opposite result. The number and morphology of neuronal mitochondria had distinct changes when compared with that of the model and NBP group. The expression rate of Bcl-2 protein in the NBP treated group was obviously higher than that in the model group. Cyt-C was weakly expressed in nerve cells of the normal and the NBP group, but was strongly expressed in the model group.ConclusionsDifferent doses of NBP administration can produce different effects. NBP at a dose of lOμmol/L may be the optimal dose. NBP treatment can induce the expression of Bcl-2 protein and decrease the release of Cyt-C from mitochondria, thereby NBP can attenuate cell death and protect cell. At the same time, NBP may inhibite the activity of MDA, which can reduce lipid peroxidation, active the SOD,which can scavenge oxygen free radicals, thereby NBP plays a role in the protection of mitochondria.Our conclusion would open a new window for the clinical treatment of Alzheimer disease.