Activation of muscarinic receptors inhibits glutamate-induced GSK-3β overactivation in PC12 cells  

作　　者：Ke MA Li-min YANG Hong-zhuan CHEN Yang LU 

机构地区：[1]Department of Pharmacy, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

出　　处：《Acta Pharmacologica Sinica》2013年第7期886-892,共7页中国药理学报（英文版）

摘　　要：Aim： To investigate the actions of the muscarinic agonist carbachol on glutamate-induced neurotoxicity in PC12 cells, and the underlying mechanisms. Methods： PC12 cells were treated with different concentrations of glutamate for 24 or 48 h. The cell viability was measured using MTT assay, and the expression and activation of GSK-3β were detected with Western blot. 13-Catenin translocation was detected using immunofluorescence. Luciferase reporter assay and real-time PCR were used to analyze the transcriptional activity of β-catenin. Results： Glutamate （1, 3, and 10 mmol/L） induced PC12 cell death in a dose-dependent manner. Moreover, treatment of the cells with glutamate （1 mmol/L） caused significant overactivation of GSK-3β and prevented β-catenin translocation to the nucleus. Pretreatment with carbachol （0.01 pmol/L） blocked glutamate-induced cell death and GSK-3β overactivation, and markedly enhanced β-catenin transcriptional activity. Conclusion： Activation of muscarinic receptors exerts neuroprotection in PC12 cells by attenuating glutamate-induced GSK-3β overactivation, suggesting potential benefits of muscarinic agonists for Alzheimer＇s disease.Aim： To investigate the actions of the muscarinic agonist carbachol on glutamate-induced neurotoxicity in PC12 cells, and the underlying mechanisms. Methods： PC12 cells were treated with different concentrations of glutamate for 24 or 48 h. The cell viability was measured using MTT assay, and the expression and activation of GSK-3β were detected with Western blot. 13-Catenin translocation was detected using immunofluorescence. Luciferase reporter assay and real-time PCR were used to analyze the transcriptional activity of β-catenin. Results： Glutamate （1, 3, and 10 mmol/L） induced PC12 cell death in a dose-dependent manner. Moreover, treatment of the cells with glutamate （1 mmol/L） caused significant overactivation of GSK-3β and prevented β-catenin translocation to the nucleus. Pretreatment with carbachol （0.01 pmol/L） blocked glutamate-induced cell death and GSK-3β overactivation, and markedly enhanced β-catenin transcriptional activity. Conclusion： Activation of muscarinic receptors exerts neuroprotection in PC12 cells by attenuating glutamate-induced GSK-3β overactivation, suggesting potential benefits of muscarinic agonists for Alzheimer＇s disease.

关 键 词：muscarinic receptors CARBACHOL neuroprotection GLUTAMATE EXCITOTOXICITY GSK-313 13-catenin PC12 cell Alzheimer＇s dis-ease 

分 类 号：Q421[生物学—神经生物学] Q425[生物学—生理学]