TAT-GluA2CT干扰GluA2/TARPγ-8耦合对癫痫持续状态模型大鼠神经损伤的保护作用  

TAT-GluA2CT interferes with the protective effect of GluA2/TARPγ-8 coupling on nerve injury in status epilepticus model of rats

作　　者：王梦露 王英燕 王纪文[1] Wang Menglu;Wang Yingyan;Wang Jiwen(Department of Neurology,Shanghai Children′s Medical Center,School of Medicine,Shanghai Jiao Tong University,Shanghai 200127,China)

机构地区：[1]上海交通大学医学院附属上海儿童医学中心神经内科,上海200127

出　　处：《中华实用儿科临床杂志》2022年第11期855-860,共6页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(8177051141)。

摘　　要：目的探究应用干扰肽TAT-GluA2CT对氯化锂-匹罗卡品癫痫持续状态模型大鼠海马神经元的保护作用以及最合适的给药时间。方法应用氯化锂-匹罗卡品建立大鼠癫痫持续状态模型(雄性, 72只), 同时设立对照组(12只)。采用随机数字表法将大鼠分为癫痫组(12只), 对照肽组(12只), 干扰肽组(48只), 根据给药的时间不同将干扰肽组又分为前1 h组(12只), 后2 h组(12只), 后4 h组(12只)和后6 h组(12只)。选择对照组、对照肽组、前1 h组、后2 h组、后4 h组、后6 h组各6只, 采用Nissl染色、原位末端标记(TUNEL)染色观察海马CA1区神经元形态变化、凋亡发生;选择对照组、对照肽组、前1 h组、后2 h组、后4 h组、后6 h组各6只, 采用Western blot和免疫共沉淀实验检测α-氨基-3-羟基-5-甲基异唑-4-丙酸(AMPA)受体第二亚单位GluA2表达和GluA2/AMPA受体胯膜调节蛋白家族γ-8(TARPγ-8)复合物耦合的变化情况。采用t检验比较2组间数据差异, 用单因素方差分析进行各组间差异的比较。结果应用干扰肽后, 与癫痫组相比, 各干扰肽组神经元数量明显增加, 差异有统计学意义(癫痫组20.07±3.51, 前1 h组39.40±2.39, 后2 h组38.43±2.42, 后4 h组30.30±2.55, 后6 h组27.93±3.20, F=235.28, P<0.05);各干扰肽组与癫痫组相比, 凋亡细胞数量明显减少, 差异有统计学意义(癫痫组31.47±3.19, 前1 h组7.30±3.45, 后2 h组9.27±3.81, 后4 h组12.86±3.08, 后6 h组14.43±3.13, F=248.60, P<0.05);与对照组相比, 诱导癫痫后海马GluA2表达量减少, 差异有统计学意义(对照组21 626.53±2 700.58, 癫痫组14 578.16±2 917.02, 前1 h组13 375.47±3 180.54, 后2 h组15 244.10±1 390.41, 后4 h组15 799.16±4 559.49, 后6 h组15 722.95±1 756.01, F=3.83, P<0.05), 各干扰肽组与癫痫组之间GluA2表达量差异无统计学意义(F=0.45, P=0.77);与癫痫组相比, 各干扰肽组GluA2/TARPγ-8耦合减少, 差异有统计学意义(癫痫组24 509.80±3 718.54, 前Objective To investigate the protective effect of interfering peptide TAT-GluA2CT on hippocampal neurons in the Lithium chlorine-Pilocarpine status epilepticus model and the optimal time of administration.Methods Male SD rats(72 cases)were induced to status epilepticus by using Lithium chlorine-Pilocarpine,while a control group(n=12)was established.The 72 rats were divided into epilepsy group(n=12),TAT-sham peptide group(n=12),TAT-GluA2CT peptide group(n=48)according to the random number table method,and the TAT-GluA2CT peptide group were further divided into the pre-1 h group(n=12),the post-2 h group(n=12),the post-4 h group(n=12),and the post-6 h group(n=12)according to the administration time of the TAT-GluA2CT peptide.Nissl staining and terminal dUTP nick end labeling(TUNEL)assay were performed on 6 rats each from control group,epilepsy group,TAT-shampeptide group,pre-1 h group,post-2 h group,post-4 h group,and post-6 h group to observe the morphological changes and apoptosis of neurons in the CA1 region of the rat hippocampus.Western blot and co-immunopercipitation test were used to detect the expression of GluA2[second subunit ofα-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid(AMPA)recepter]and the coupling of GluA2/transmembrane AMPA receptor regulatory protein(TARPγ-8)complex in control group,epilepsy group,pre-1 h group,post-2 h group,post-4 h group and post-6 h group.The t-test was used to compare the data differences between 2 groups,and one-way ANOVA was adopted to compare the differences between the groups.Results Compared with the epilepsy group,the number of neurons in each TAT-GluA2CT peptide group increased significantly,and the difference was statistically significant(epilepsy group 20.07±3.51,pre-1 h group 39.40±2.39,post-2 h group 38.43±2.42,post-4 h group 30.30±2.55,and post-6 h group 27.93±3.20,F=235.28,P<0.05).Compared with the epilepsy group,the number of apoptotic cells in each TAT-GluA2CT peptide group was significantly reduced,and the difference was statistically significant

关 键 词：癫痫持续状态 α-氨基-3-羟基-5-甲基异唑-4-丙酸受体 AMPA受体跨膜调节蛋白家族 GluA2/TARPγ-8复合物 

分 类 号：R742.1[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]