基于"多重打击"理论建立SD大鼠认知功能障碍模型及相关机制研究  

"Multi-hits"based SD rat models of cognitive dysfunction and research on related mechanisms

作　　者：段博阳 王月 杨李[2] 刘乐 崔珍珍 余莉 唐久来[2] 吴德 Duan Boyang;Wang Yue;Yang Li;Liu Le;Cui Zhenzhen;Yu Li;Tang Jiulai;Wu De(Department of Pediatrics,the Fourth Affiliated Hospital of Anhui Medical University,Hefei 230000,China;Department of Pediatrics,the First Affiliated Hospital of Anhui Medical University,Hefei 230000,China;Anhui Provincial Laboratory of Microbiology and Parasitology,Hefei 230000,China)

机构地区：[1]安徽医科大学第四附属医院儿科,合肥230000 [2]安徽医科大学第一附属医院儿科,合肥230000 [3]病原生物学安徽重点实验室,合肥230000

出　　处：《中华实用儿科临床杂志》2022年第13期1011-1016,共6页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(81472167)。

摘　　要：目的通过脑发育早期脂多糖(LPS)重复刺激,建立SD大鼠认知障碍模型,探讨炎症反应介导下的"多重打击"对其组织学、行为学的影响及相关的分子机制。方法本研究的实验设计类型为成组设计。通过腹腔注射LPS构建SD大鼠"多重打击"认知障碍模型,将24只孕鼠按照随机数字表法分为对照组、LPS1组、LPS2组、LPS3组,每组6只。对照组孕18 d孕鼠和20日龄仔鼠均腹腔注射9 g/L盐水;LPS1组孕18 d孕鼠腹腔注射9 g/L盐水;LPS2组孕18 d孕鼠腹腔注射0.05 mg/kg LPS;LPS3组孕18 d孕鼠注射0.1 mg/kg LPS。LPS1~3组仔鼠均在20日龄时腹腔注射1 mg/kg LPS。通过前肢悬吊、网格和水迷宫等行为学实验,对仔鼠运动和认知功能进行整体评估。组织学和蛋白免疫印迹(WB)实验检测仔鼠脑组织中胶质纤维酸性蛋白(GFAP)、Notch1和Jagged1的相对表达量。采用单因素方差分析进行多组数据之间比较,采用独立样本t检验进行2组数据之间比较。结果1.行为学实验:与对照组比较,LPS1~3组仔鼠的前肢悬吊时间递减[(34.81±5.66)s、(22.47±4.35)s、(13.20±4.25)s比(43.88±4.85)s],网格实验中足踏空次数递增(16.13±2.90、20.75±3.10、25.13±4.45比9.00±2.72),差异均有统计学意义(F=69.77、35.59,均P<0.001);Morris水迷宫实验中的潜伏期时间和总路程呈递增趋势(P<0.05)。2.WB实验:与对照组比较,LPS1~3组GFAP、Notch1、Jagged1蛋白的相对表达量显著增加(P<0.05)。3.苏木精-伊红(HE)染色和电镜病理:与对照组比较,LPS1~3组仔鼠额叶皮质组织结构排列松散,细胞固缩明显。电镜下细胞质不同程度水肿、线粒体嵴断裂、部分核膜破损。结论在"多重打击"认知障碍模型中,仔鼠脑组织的损伤和行为学的改变,可能与LPS重复暴露介导的Notch1/Jagged1通路上调有关。Objective To establish Sprague-Dawley(SD)rat models of cognitive impairment through repeated stimulation of lipopolysaccharide(LPS)in the early brain development,and to inquire into the effect of"multi-hits"mediated by inflammatory response on the histology and behavior of SD rat models and related molecular mechanisms.Methods This study adopted a group design for experiments.The"multi-hits"SD rat models were established by intraperitoneal injection of LPS.According to the random number table method,24 pregnant rats were randomly divided into 4 groups:control group,LPS1 group,LPS2 group and LPS3 group,6 rats in each group.In the control group,saline was intraperitoneally injected into rats with gestational age of 18 days and 20-day-old neonatal rats.Rats with gestational age of 18 days were intraperitoneally injected with saline in the LPS1 group,0.05 mg/kg LPS in the LPS2 group,and 0.1 mg/kg LPS in the LPS3 group.The pups in LPS1-3 groups were all injected intraperitoneally with 1 mg/kg LPS at the postnatal age of 20 days.The motor and cognitive function of the pups were evaluated overall by behavioral experiments such as forelimb suspension tests,grid tests and water maze tests.The relative expression of glial fibrillary acidic protein(GFAP),Notch1 and Jagged1 in brain tissue of pups was mainly detected by Western blot(WB)and histological experiments.One-way ANOVA analysis of variance and independent samples t-test were used to compare data among groups and between groups,respectively.Results(1)Behavioral experiments:compared with the control group,LPS1-3 groups showed progressive decrease in forelimb suspension time[(34.81±5.66)s,(22.47±4.35)s,and(13.20±4.25)s vs.(43.88±4.85)s],and the number of missteps in the grid experiment increased progressively(16.13±2.90,20.75±3.10,25.13±4.45 vs.9.00±2.72).The differences were statistically significant(F=69.77,35.59,all P<0.001).Both the escape latency and total distance in Morri′s water maze test increased progressively(P<0.05).(2)WB experiment:the relative e

关 键 词：脑组织 脂多糖 认知障碍 星形胶质细胞 胶质纤维酸性蛋白 Notch1/Jagged1信号通路 SD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]