基于PI3K/AKT/GSK-3β信号通路探讨酸枣仁汤对APP/PS1双转基因痴呆小鼠海马神经元突触损伤的改善作用  

Discussion on the effect of Suanzaoren Decoction on hippocampal neurons and synaptic damage in APP/PS1 double transgenic dementia mice based on PI3K/AKT/GSK-3βsignaling pathway

作　　者：龙清华 赵宾宾 丁莉 徐波 谭爱华 王平 LONG Qing-hua;ZHAO Bin-Bin;DING Li;XU Bo;TAN Ai-hual;WANG Ping(School of Basic Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;institute of Geriatrics,Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学基础医学院,武汉430065 [2]湖北中医药大学老年医学研究所,武汉430065

出　　处：《中华中医药杂志》2020年第5期2546-2551,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81573865)。

摘　　要：目的:观察酸枣仁汤对APP/PS1小鼠海马磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)/糖原合成酶激酶3β(GSK-3β)信号通路的作用,并探讨其改善学习记忆和神经元-突触损伤的可能机制。方法:将小鼠随机分为空白(CON)组、模型(MOD)组、多奈哌齐(DON)组和酸枣仁汤低剂量(L-SZRD)组和酸枣仁汤高剂量(H-SZRD)组,并给予相应药物治疗。采用Morris水迷宫评价小鼠的学习记忆力,采用尼氏染色和高尔基染色观察小鼠海马神经元和突触损伤情况,采用免疫荧光检测小鼠海马中突触后致密蛋白(PSD95)和突触素(SYN)的蛋白表达水平,采用Western blot检测小鼠海马中Tau蛋白及其磷酸化蛋白表达水平,并对PI3K/AKT/GSK-3β信号通路中关键蛋白及其磷酸化蛋白表达水平进行检测。结果:与CON组比较,MOD组小鼠平台潜伏期显著增加(P<0.01),穿越平台次数和目标象限停留时间显著减少(P<0.01);海马CA3区神经元数量、树突棘密度和树突分支数显著减少(P<0.01);海马CA3区PSD95和SYN明显下降;海马中Tau蛋白相关磷酸化蛋白表达水平显著升高(P<0.01);海马中pY607-PI3K、pS473-AKT和pS9-GSK-3β的蛋白表达水平显著降低(P<0.01)。与MOD组比较,各给药组小鼠平台潜伏期减少(P<0.01,P<0.05),而穿越平台次数和目标象限停留时间增加(P<0.01,P<0.05);3组小鼠海马CA3区神经元数量、树突棘密度和树突分支数均有增加(P<0.05),CA3区PSD95和SYN有所增加;海马中Tau蛋白相关磷酸化蛋白表达水平降低(P<0.05);海马中pY607-PI3K、pS473-AKT和pS9-GSK-3β的蛋白表达水平升高(P<0.05,P<0.01)。结论:酸枣仁汤可以改善APP/PS1痴呆小鼠的学习记忆力,并能抑制神经元和突触损伤,其机制可能与其激活PI3K/AKT/GSK-3β信号通路和抑制Tau蛋白磷酸化有关。Objective: To observe the effect of Suanzaoren Decoction on PI3 K/AKT/GSK-3β signaling pathway in hippocampus of APP/PS1 mice, and to explore its possible mechanism of improving learning and memory and neuron-synaptic damage. Methods: The mice were randomly divided into CON group, MOD group, DON group, L-SZRD group and H-SZRD group, and given corresponding drug. The learning and memory of mice were evaluated by Morris water maze. The hippocampal neurons and synaptic damage were observed by Nissl staining and Golgi staining. The expression levels of PSD95 and SYN in hippocampus of mice were detected by immunofluorescence. Western blot was used to detect the expression levels of Tau protein and its phosphorylated protein in mouse of hippocampus. The expression levels of key proteins and their phosphorylated proteins in PI3 K/AKT/GSK-3β signaling pathway were detected by Western blot. Results: Compared with the CON group, the escape latency of the mouse platform in the MOD group increased significantly(P<0.01), and the number of crossing platforms and the target quadrant residence time were significantly reduced(P<0.01). Compared with the CON group, the number of neurons, dendritic spine density and dendritic branch number in the hippocampal CA3 area of the MOD group were significantly decreased(P<0.01). Compared with the CON group, PSD95 and SYN in the hippocampal CA3 area of the MOD group were significantly decreased. Compared with the CON group, the expression of Tau-associated phosphorylated protein in the hippocampus of the MOD group was significantly increased(P<0.01). Compared with the CON group, the protein expression levels of pY607-PI3 K, pS473-AKT and pS9-GSK-3β in hippocampus of MOD group were significantly decreased(P<0.01). Compared with the MOD group, escape latency of the DON group, the L-SZRD group and the H-SZRD group was decreased(P<0.01, P<0.05), while the number of crossing platforms and target quadrant retention time were increased(P<0.01, P<0.05). Compared with MOD group, the number of neur

关 键 词：酸枣仁汤 阿尔茨海默病 痴呆 学习记忆力 神经元 突触 机制 磷脂酰肌醇3激酶/蛋白激酶B/糖原合成酶激酶3β信号通路 Tau蛋白 

分 类 号：R285.5[医药卫生—中药学]