摘要
目的探讨鞣花酸(ellagicacid,EA)对APP/PS1双转基因小鼠认知功能的影响,并基于磷脂酰肌醇3-激酶/蛋白激酶B/糖原合成酶激酶-3(PI3K/AKT/GSK-3β)信号通路探讨鞣花酸对双转基因小鼠海马氧化应激水平的调节机制。方法将32只SPF级6月龄APP/PS1双转基因小鼠随机分为4组,即APP/PS1组、APP/PS1+EA组、APP/PS1+LY294002组、APP/PS1+EA+LY294002组,每组8只,另外选取8只SPF级C57BL/6J野生型小鼠(Wildtype)作为空白对照组,即WT组。APP/PS1+EA组给予50mg·kg^(-1)·d^(-1)灌胃EA;APP/PS1+LY294002组予以1.5mg·kg^(-1)·d^(-1)腹腔注射PI3K抑制剂LY294002;APP/PS1+EA+LY294002组予以50mg·kg^(-1)·d^(-1)灌胃EA,同时按1.5mg·kg^(-1)·d^(-1)腹腔注射LY294002;WT组和APP/PS1组于相同时间点灌胃等体积10%二甲基亚砜(DMSO)。每日给药1次,连续给药60天。Morris水迷宫检测小鼠学习和记忆能力,免疫组化、蛋白免疫印迹法检测PI3K、AKT、GSK-3β相关蛋白的表达,透射电镜观察小鼠海马组织超微结构变化。结果与WT组相比,其他四组的逃避潜伏期均增长(P<0.05),穿越平台次数明显减少(P<0.01);APP/PS1组、APP/PS1+LY294002组和APP/PS1+EA+LY294002组中的PI3K、AKT蛋白表达量显著降低(P<0.01),GSK-3β表达量显著升高(P<0.01);APP/PS1+EA组的PI3K表达量降低(P<0.05),AKT表达量显著降低(P<0.01),GSK-3β表达量升高(P<0.05);与WT组相比,APP/PS1组海马神经元细胞数目较少,线粒体结构破坏,大部分线粒体出现肿胀,线粒体的内膜和外模不完整,部分线粒体嵴消失,微管、微丝缠结,排列紊乱,而APP/PS1+EA组神经元细胞数较APP/PS1组增多,线粒体结构较清晰,可见清楚的线粒体嵴,线粒体轻度水肿。微管、微丝排列较整齐有序。结论鞣花酸改善AD模型小鼠的学习和记忆能力、减少海马神经元细胞损伤和凋亡,其作用机制可能是通过调节PI3K、AKT、GSK-3β等相关蛋白降低AD模型小鼠海马氧化应激水平。
Aim To investigate the effect of ellagic acid(EA)on cognitive function in APP/PS1 double-transgenic mice,and to explore the regulatory mechanism of ellagic acid on the level of oxidative stress in the hippocampus of double-transgenic mice based on the phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase-3(PI3K/AKT/GSK-3β)signaling pathway.Methods Thirty-two SPF-grade 6-month-old APP/PS1 double transgenic mice were randomly divided into four groups,namely,APP/PS1 group,APP/PS1+EA group,APP/PS1+LY294002 group,APP/PS1+EA+LY294002 group,with eight mice in each group,and eight SPF-grade C57BL/6J wild type mice(Wild type)were selected as the blank control group.The APP/PS1+EA group was given 50 mg·kg^(-1)·d^(-1) of EA by gavage;the APP/PS1+LY294002 group was given 1.5 mg·kg^(-1)·d^(-1) of PI3K inhibitor LY294002 by intraperitoneal injection;the APP/PS1+EA+LY294002 group was given 50 mg·kg^(-1)·d^(-1) of EA by gavage,and the APP/PS1+EA+LY294002 group was given 50 mg·kg^(-1)·d^(-1) of EA by gavage.The WT group and APP/PS1 group were given equal volume of 10%dimethyl sulfoxide(DMSO)by gavage at the same time.Morris water maze was administered once a day for 60 days,and the learning and memory abilities of mice were measured by immunohistochemistry and protein immunoblotting(WB),and the expressions of PI3K,AKT and GSK-3β-related proteins were detected by transmission electron microscopy.Results Compared with the WT group,the other four groups all showed an increase in escape latency(P<0.05)and a significant decrease in the number of plateaus crossed(P<0.01);PI3K and AKT protein expression was significantly lower in the APP/PS1 group,APP/PS1+LY294002 group and APP/PS1+EA+LY294002 group(P<0.01),GSK-3βexpression was significantly higher(P<0.01);PI3K expression was lower(P<0.05),AKT expression was significantly lower(P<0.01)and GSK-3βexpression was higher(P<0.05)in the APP/PS1+EA group;compared with the WT group,the APP/PS1 group had a lower number of hippocampal neuronal cells,disrupted mitochon
作者
仲丽丽
路鑫
于颖
赵秦妍
张静
刘彤慧
倪雪妍
车艳玲
吴丹
刘宏
ZHONG Li-li;LU Xin;YU Ying;ZHAO Qin-yan;ZHANG Jing;LIU Tong-hui;NI Xue-yan;CHE Yan-ling;WU Dan;LIU Hong(Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,China;the First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China;Dept of Pathology,School of Basic Sciences,Heilongjiang University of ChineseMedicine,Harbin 150040,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第1期90-98,共9页
Chinese Pharmacological Bulletin
基金
国家自然科学基金青年项目(No81403288)
国家自然科学基金面上项目(No82074288)
黑龙江省自然科学基金联合引导项目(NoLH2022H068)
中国博士后科学基金资助课题(No2015M581496)
黑龙江中医药大学研究生创新科研项目立项基金(No2022yjscx012,2019yjscx011)
黑龙江省卫生健康委科研课题立项(No20220202080996)。
