Bian Hong, Bian Wei, Lin Xiaoying, Ma Zhaoyin, Chen Wen, Pu Ying
Department of Neurology, Jinan Central Hospital, Shandong University School of Medicine, No. 105 Jiefang Road, Jinan, Shandong, China.
Department of Endocrinology, The Fourth People's Hospital of Jinan, Jinan, China.
Neurochem Res. 2016 Sep;41(9):2470-80. doi: 10.1007/s11064-016-1960-7. Epub 2016 Jun 2.
To explore the effect of glycogen synthase kinase 3β (GSK-3β) silencing on Tau-5 phosphorylation in mice suffering Alzheimer disease (AD). GSK-3β was firstly silenced in human neuroblastoma SH-SY5Y cells using special lentivirus (LV) and the content of Tau (A-12), p-Tau (Ser396) and p-Tau (PHF-6) proteins. GSK-3β was also silenced in APP/PS1 mouse model of AD mice, which were divided into three groups (n = 10): AD, vehicle, and LV group. Ten C57 mice were used as control. The memory ability of mice was tested by square water maze, and the morphological changes of hippocampus and neuron death were analyzed by haematoxylin-eosin staining. Moreover, the levels of Tau and phosphorylated Tau (p-Tau) were detected by western blotting and immunohistochemistry, respectively. The lentivirus-mediated GSK-3β silencing system was successfully developed and silencing GSK-3β at the cellular level reduced Tau phosphorylation obviously. Moreover, GSK-3β silence significantly improved the memory ability of AD mice in LV group compared with AD group (P < 0.05) according to the latency periods and error numbers. As for the hippocampus morphology and neuron death, no significant change was observed between LV group and normal control. Immunohistochemical detection and western blotting revealed that the levels of Tau and p-Tau were significantly down-regulated after GSK-3β silence. Silencing GSK-3β may have a positive effect on inhibiting the pathologic progression of AD through down-regulating the level of p-Tau.
探讨糖原合酶激酶3β（GSK-3β）沉默对阿尔茨海默病（AD）小鼠Tau-5磷酸化的影响。首先使用特殊慢病毒（LV）在人神经母细胞瘤SH-SY5Y细胞中沉默GSK-3β，并检测Tau（A-12）、p-Tau（Ser396）和p-Tau（PHF-6）蛋白的含量。还在AD小鼠的APP/PS1小鼠模型中沉默GSK-3β，将其分为三组（n = 10）：AD组、载体组和LV组。以10只C57小鼠作为对照。通过方形水迷宫测试小鼠的记忆能力，并用苏木精-伊红染色分析海马的形态变化和神经元死亡情况。此外，分别通过蛋白质印迹法和免疫组织化学检测Tau和磷酸化Tau（p-Tau）的水平。成功构建了慢病毒介导的GSK-3β沉默系统，在细胞水平沉默GSK-3β可明显降低Tau磷酸化。此外，根据潜伏期和错误次数，与AD组相比，LV组中GSK-3β沉默显著改善了AD小鼠的记忆能力（P < 0.05）。至于海马形态和神经元死亡情况，LV组与正常对照组之间未观察到显著变化。免疫组织化学检测和蛋白质印迹法显示，GSK-3β沉默后Tau和p-Tau的水平显著下调。沉默GSK-3β可能通过下调p-Tau水平对抑制AD的病理进展产生积极作用。
