自闭症模型小鼠神经递质水平病理性改变的机制研究  

Mechanism of pathological changes in neurotransmitter levels in a mouse autism model

作　　者：赵佳佳[1] 赵蓓[1] 高未 李杰 蔡俏[1] 应小明[1] ZHAO Jiajia;ZHAO Bei;GAO Wei;LI Jie;CAI Qiao;YING Xiaoming(Department of Pediatrics,Taizhou First People's Hospital,Taizhou 318020,China)

机构地区：[1]台州市第一人民医院儿科,318020

出　　处：《浙江医学》2022年第4期350-354,共5页Zhejiang Medical Journal

基　　金：浙江省科技发展专项资金(2018C37106)。

摘　　要：目的探讨自闭症模型小鼠神经递质水平病理性改变的机制。方法取健康成年雌鼠12只和雄鼠6只合笼,受孕后将雌鼠按随机数字表法分为模型组(母代)和对照组(母代),其中模型组(母代)于受孕10、12 d腹腔注射丙戊酸钠(VPA)注射液(使用0.9%氯化钠注射液配置,浓度为250 mg/ml)1.2 ml/kg,对照组给予等体积0.9%氯化钠注射液。模型组小鼠(子代)出生4 d后进行母子分离24 h后送回同笼,建立自闭症小鼠模型。建模8周后,取模型组(子代)、对照组(子代)各12只,其中各取3只采用理毛实验、水迷宫实验和旷场实验观察其行为学改变;各取3只采用高尔基染色法观察海马锥体神经元结构;各取3只采用高效液相色谱法测定脑皮层γ-氨基丁酸(GABA)、5-羟色胺(5-HT)和多巴胺水平;各取3只采用Western blot法测定脑组织Arc、EGR2和S-100a9蛋白表达水平。结果与对照组小鼠(子代)比较,模型组小鼠(子代)理毛次数明显增多(P<0.05),第2、3、4、5天潜伏时间均明显延长(均P<0.05),总运动距离明显缩短(P<0.05),穿越格子次数、后肢站立次数均明显减少(均P<0.05);锥体神经元数量明显减少(P<0.05);脑皮层GABA、5-HT水平均明显升高(均P<0.05),而多巴胺水平明显降低(P<0.05);脑组织Arc、EGR2蛋白表达水平均明显升高(均P<0.05),而S-100a9蛋白表达水平明显降低(P<0.05)。结论自闭症模型小鼠存在行为学异常和神经递质水平改变,可能与Arc、EGR2蛋白表达上调、S-100a9蛋白表达下调的机制有关。Objective To investigate the mechanism of pathological changes in neurotransmitter levels in a mouse autism model.Methods Twelve healthy adult female rats and six male rats were caged in a cage.The pregnant mice were randomly divided into two groups.The maternal model group was given intraperitoneal injection of 1.2 ml/kg sodium valproate(VPA,250 mg/ml)on the 10th and 12th day of conception,and the control group was given equal volume of normal saline.Twelve offspring mice were taken from each group to set offspring model and control groups.The offspring model group was subjected to a mother-child separation for 4 days after birth to induce autism,while no special treatment was given for offspring control group.The behavior changes were observed with water maze test,grooming and open field experiments.The structure and morphology of hippocampal neurons was observed with Golgi staining.The levels ofγ-aminobutyric acid(GABA),5-hydroxytryptamine(5-HT)and dopamine in brain were determined with high performance liquid chromatography.The protein expression of Arc,EGR2 and S-100a9 was measured by Western blot.Results The mice in the model group had more grooming times than that in the control group(P<0.05).The latent period of the model group was significantly longer than that of the control group at d2 to d5(all P<0.05).The total movement distance of mice in the model group was significantly shorter than that of the control group(P<0.05),and the number of times of crossing grids and standing times of hind legs was significantly less than that of the control group(both P<0.05).Compared with the control group,the hippocampal neuron cell bodies of the model group were atrophied and the number of vertebral neurons decreased(P<0.05).The GABA and 5-HT levels of the model group were higher than those of the control group(both P<0.05),while the dopamine level was lower than that of the control group(P<0.05).The expression of Arc and EGR2 protein in the model group was higher than that in the control group(both P<0.05),while t

关 键 词：自闭症 神经递质 ARC EGR2 S-100a9 

分 类 号：R749.94[医药卫生—神经病学与精神病学]