两种海人酸癫痫小鼠模型的建立及比较研究  

Establishment and comparison of 2 mouse models of kainic acid induced epilepsy

作　　者：滕灿 余美玲 蒋国会 范会业[3] 王法祥 TENG Can;YU Meiling;JIANG Guohui;FAN Huiye;WANG Faxiang(Department of Neurology,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037;Department of Neurology,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan Province,637000,China;Clinical Medical Research Center,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037)

机构地区：[1]陆军军医大学(第三军医大学)第二附属医院神经内科,重庆400037 [2]川北医学院附属医院神经内科,四川南充637000 [3]陆军军医大学(第三军医大学)第二附属医院临床医学研究中心,重庆400037

出　　处：《陆军军医大学学报》2024年第7期661-669,共9页Journal of Army Medical University

基　　金：国家自然科学基金面上项目(81971221);重庆市自然科学基金面上项目(cstc2021jcyj-msxm3511)。

摘　　要：目的比较分析海人酸(kainic acid,KA)侧脑室(intracerebroventricular,ICV)注射和腹腔注射(intraperitoneal injections,IP)两种给药路径建立的颞叶癫痫小鼠模型早期的行为学和病理学差异。方法100只C57BL/6N野生型(wild type,WT)雄性小鼠(体质量为20~22 g),按随机数字表法分为4组:ICV+normal saline(NS)对照组(n=10),ICV+KA模型组(n=40),IP+NS对照组(n=10),IP+KA模型组(n=40)。ICV+KA模型组使用600 nL的KA(0.5 mg/mL)进行侧脑室注射,IP+KA模型组按25 mg/kg剂量进行腹腔注射,对照组使用等量的生理盐水进行侧脑室和腹腔注射。建立模型3 d后,进行行为学、分子生物学(Western blot)和神经病理损伤评估(FJB染色,TUNEL染色,免疫荧光染色)。结果两对照组无痫性发作,两模型组均出现痫性发作。IP+KA组与ICV+KA组死亡率分别为47.50%和65.00%,造模成功率分别为80.00%与60.00%。与IP+KA组比较,ICV+KA组小鼠模型成功率明显增高而死亡率明显减少。FJB染色和TUNEL染色结果显示,与IP+KA组比较,ICV+KA组海马的神经变性和凋亡改变程度变化更加明显(P<0.05)。与IP+KA组比较,ICV+KA组海马凋亡蛋白表达差异也有统计学意义(P<0.05)。免疫荧光结果显示,ICV+KA和IP+KA组海马和皮层的星形胶质细胞和小胶质细胞较对照组明显激活(P<0.05),但是ICV+KA组海马和皮层的胶质细胞激活程度均强于IP+KA组(P<0.05)。ICV+KA组海马和皮层的GFAP和Iba-1蛋白表达均高于IP+KA组(P<0.05)。结论两种KA制备方法均可制备出成功的癫痫模型。但ICV路径制备癫痫小鼠具有更高的成功率和更少的死亡率,并且神经病理损害程度和胶质细胞激活程度更加明显。Objective To investigate and analyze the behavioral and pathological differences in early-stage mouse models of epilepsy established by 2 different administration routes for kainic acid(KA),intracerebroventricular(ICV)injection and intraperitoneal(IP)injection.Methods A total of 100 male C57BL/6N wild-type(WT)mice(20~22 g)were randomly divided into ICV+normal saline(NS)control group(n=10),ICV+KA model group(n=40),IP+NS control group(n=10)and IP+KA model group(n=40).The ICV+KA model group was given 600 nL of KA(0.5 mg/mL)via ICV injection,and the IP+KA model group was injected with different dose of KA(25 mg/kg).Two control groups were administered equal volumes of NS via corresponding routes.After 3 d of modeling,the evaluation of behavioristics,molecular biology(including Western blotting),and neuropathological assessments(including FJB staining,TUNEL staining and immunofluorescence staining)were performed.Results No epileptic seizures were observed in both 2 control groups,while exhibited seizures were observed in both model groups.The mortality rates of the IP+KA group and the ICV+KA group were 47.50%and 65.00%respectively,while the success rates of modeling were 80.00%and 60.00%respectively.Compared with the IP+KA group,the ICV+KA group showed a significant increase in success rate and a significant reduction in mortality rate.FJB and TUNEL staining results showed that,compared with the IP+KA group,the severity of neurodegeneration and apoptotic changes in the hippocampus of the ICV+KA group were more significant(P<0.05).Compared with the IP+KA group,there was also a significant difference in the expression of apoptotic proteins in the hippocampus of the ICV+KA group(P<0.05).Immunofluorescence results showed that the astrocytes and microglia in the hippocampus and cortex of the ICV+KA and IP+KA groups were significantly activated compared with the control groups(P<0.05),but the activation of glial cells in the hippocampus and cortex of the ICV+KA group was stronger than that of the IP+KA model group(P<0.05)an

关 键 词：颞叶癫痫 海人酸 侧脑室注射 腹腔注射 病理学差异 

分 类 号：R-334[医药卫生] R742.1