AMPA受体内化抑制剂GluA2-3Y对慢性脑低灌注模型大鼠认知功能及突触后蛋白表达的影响  

Effect of AMPA receptor endocytosis inhibitor GluA2-3Y on cognitive function and postsynaptic protein expression in chronic cerebral hypoperfusion rats

作　　者：吴丽阳 朱虹[1] 章军建[1] Wu Liyang;Zhu Hong;Zhang Junjian(Department of Neurology,Zhongnan Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院神经内科,武汉430071

出　　处：《中华行为医学与脑科学杂志》2021年第10期865-872,共8页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金项目(81771151,82071210)。

摘　　要：目的探究α-氨基-3-羟基-5-甲基-4-异恶唑-丙酸(α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid, AMPA)受体内化抑制剂GluA2-3Y对慢性脑低灌注大鼠认知功能及海马突触后蛋白表达的影响。方法 48只成年雄性SD大鼠按照随机数字表法分为Sham组、2VO组、高剂量GluA2-3Y组和低剂量GluA2-3Y组, 每组12只。采用双侧颈总动脉永久性结扎法(two vessel occlusion, 2VO)构建慢性脑低灌注模型, 其中Sham组行假手术。高剂量GluA2-3Y组和低剂量GluA2-3Y组分别腹腔注射剂量为3 μmol/kg和0.03 μmol/kg的GluA2-3Y, 1次/d, 共2周, 2VO组和Sham组大鼠腹腔注射对照肽。采用Morris水迷宫和新物体识别实验评测大鼠的学习记忆能力, 免疫印迹法评测大鼠海马Akt1、GSK3β(glycogen synthase kinase-3β)、p-GSK3β、GluA2和PSD-95(postsynaptic density-95)的表达, 免疫荧光法评测大鼠海马GluA2和PSD-95的表达。采用SPSS 23.0进行数据分析, 组间比较采用单因素方差分析, Morris水迷宫结果采用重复测量方差分析, 两两比较采用独立样本t检验。结果 (1)重复测量方差分析结果显示, 在Morris水迷宫实验中, 各组大鼠逃避潜伏期的分组与时间的交互作用不显著(F=0.79, P>0.05), 组别主效应与时间主效应均显著(F=24.44, 40.42, 均P<0.05)。在第5天的定位航行检测中, 2VO组大鼠的逃避潜伏期较Sham组长(t=5.87, P<0.05), 低剂量GluA2-3Y组和高剂量GluA2-3Y组大鼠的逃避潜伏期均明显短于2VO组(t=2.20, 3.41, 均P<0.05), 而高剂量GluA2-3Y组与低剂量GluA2-3Y组的逃避潜伏期差异无统计学意义(t=1.37, P>0.05)。2VO组的目标象限停留时间和分辨系数[(14.57±1.40)s, (0.15±0.10)]均明显低于Sham组[(23.71±2.57)s, (0.40±0.06)](t=3.23, 2.24, 均P<0.05), 而高剂量GluA2-3Y组[(20.19±1.53)s]和低剂量GluA2-3Y组[(20.31±2.06)s]的目标象限停留时间均较2VO组长(t=2.71, 2.35, 均P<0.05)。高剂量GluA2-3Y组(0.47±0.10)和低剂量GluA2-3Y组(0.59±0.06)�Objective To investigate the effect of GluA2-3Y which is an inhibitor of AMPA(α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid)receptor internalization on cognitive function and hippocampal postsynaptic protein expression in rats with chronic cerebral hypoperfusion.Methods Forty-eight adult male SD rats were randomly divided into Sham group,2VO group,high-dose GluA2-3Y group and low-dose GluA2-3Y group according to random number table,with 12 rats in each group.The chronic cerebral hypoperfusion model of rat was established by two vessel occlusion(2VO)while the Sham operation was performed in rats of Sham group.The rats in high dose GluA2-3Y group and low dose GluA2-3Y group were intraperitoneal injected with 3μmol/kg and 0.03μmol/kg GluA2-3Y respectively once a day for 2 weeks.Rats in 2VO group and Sham group were intraperitoneally injected with control peptide.Morris water maze test and new object recognition test were performed to evaluate the learning and memory ability of rats,and Western blot was used to evaluate the expression of Akt1、GSK3β、p-GSK3β、GluA2 and PSD-95 in rat hippocampus.The expressions of GluA2 and PSD-95 in rat hippocampus were evaluated by immunofluorescence.SPSS 23.0 software was used for data analysis.The comparison between multiple groups was analyzed by one-way ANOVA and repeated measurement ANOVA was used to analyze Morris water maze results.And independent-samples t-test was used for pairwise comparisons.Results(1)In Morris water maze trials,the results of repeated measurement ANOVA showed that the interaction between group and time of escape latency of rats in each group was not significant(F=0.79,P>0.05),and the group main effect and time main effect were significant(F=24.44,40.42,both P<0.05).On the 5th day of navigation trials,the escape latency of rats in 2VO group was longer than that in sham group(t=5.87,P<0.05).The escape latency of rats in low dose GluA2-3Y group and high dose GluA2-3Y group were significantly shorter than that in 2VO group(t=2.20,3.41,both P<

关 键 词：慢性脑低灌注 认知 AMPA受体内化 糖原合成酶激酶3Β AKT1 PSD-95 GluA2 大鼠 

分 类 号：R749.1[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]