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目的:探讨在体转染GSK-3β对tau蛋白在PHF-1位点磷酸化的影响。方法:21只大鼠随机分为GSK-3β转染组、空载体组和空白对照组3组:0.1μg/3μLGSK-3β-HA质粒和空载体分别注射入大鼠大脑,对照组大鼠不作处理,应用免疫印迹和免疫组织化学检测GSK-3β的表达,并应用磷酸化位点特异性抗体PHF-1检测tau蛋白的磷酸化水平。结果:转染48h后,GSK-3β-HA表达在转染组;并且在转染区域的神经元内异常过度磷酸化tau蛋白(在PHF-1表位)聚积;异常过度磷酸化的tau蛋白与GSK-3β共定位。结论:在体转染GSK-3β引起导致神经退行性疾病发生机制相关的tau蛋白异常过度磷酸化,这进一步证明了GSK-3β是tau蛋白异常过度磷酸化的一个关键激酶,并且可作为一个防治与tau相关的神经退行性疾病的靶点。
AIM: To investigate the effect of GSK-3β transfection on phosphorylation of tau at PHF-1 in vivo. Methods: Twenty-one rats were randomly divided into three groups: GSK-3βtransfected group, empty vector group and blank control group. 0.1μg / 3μL GSK-3β-HA plasmid and empty vector were injected into rat brain respectively. The expression of GSK-3β was detected by Western blotting and immunohistochemistry. The level of tau phosphorylation was detected by phosphorylation site-specific antibody PHF-1. Results: After 48h, GSK-3β-HA was expressed in the transfected group and abnormally hyperphosphorylated tau protein (at the PHF-1 epitope) was accumulated in the neurons in the transfected region. Abnormally hyperphosphorylated tau protein Co-localization with GSK-3β. CONCLUSIONS: In vivo transfection of GSK-3β causes abnormal hyperphosphorylation of tau associated with the pathogenesis of neurodegenerative diseases, further demonstrating that GSK-3β is a key kinase of abnormal hyperphosphorylation of tau protein and may be used as a control Targets of tau-associated neurodegenerative diseases.