基于严重痫样发作行为的小鼠匹罗卡品颞叶癫痫模型构建  

Construction of a pilocarpine-induced temporal lobe epilepsy model in mice based on severe seizure behavior

作　　者：童方超 蔡翊莹 李远方 王强 丁晶 汪昕 TONG Fangchao;CAI Yiying;LI Yuanfang;WANG Qiang;DING Jing;WANG Xin(Department of Neurology,Zhongshan Hospital,Fudan University,Shanghai 200032,China;Department of Neurology,Zhongshan Hospital(Xiamen Branch),Fudan University,Xiamen 361015,Fujian,China)

机构地区：[1]复旦大学附属中山医院神经内科,上海200032 [2]复旦大学附属中山医院厦门医院神经内科,厦门361015

出　　处：《中国临床医学》2024年第5期712-723,共12页Chinese Journal of Clinical Medicine

基　　金：国家自然科学基金面上项目(81771308,31771184,31970901);上海浦江人才计划项目(19PJ14002200)。

摘　　要：目的探讨C57BL/6J亚系小鼠经腹腔注射匹罗卡品建立颞叶癫痫模型的方法,总结可用于预测造模成功的癫痫发作急性期行为学表现,为癫痫研究提供可行的造模方案。方法选择30只C57BL/6J亚系小鼠(主要研究对象)和40只C57BL/6N亚系小鼠(对照),通过单次腹腔注射匹罗卡品的方法诱导小鼠癫痫发作从而建立颞叶癫痫模型。两种亚系的小鼠各分为3组,分别经腹腔注射300 mg/kg、330 mg/kg或360 mg/kg的匹罗卡品,观察并比较两种亚系小鼠的运动性癫痫发作的行为学表现,并在造模后第7天开始连续监测小鼠的自发性癫痫发作(spontaneous recurrent seizures,SRS)行为。造模后28 d,处死小鼠并观察其海马的病理改变情况。结果注射匹罗卡品后,C57BL/6N亚系小鼠表现出典型的运动性癫痫发作,而后进入癫痫持续状态(status epilepticus,SE);C57BL/6J小鼠较少观察到典型的运动性癫痫发作及后续的SE,而更多表现为单侧肢体抽搐后持续数秒至数十秒的全身颤抖,本研究将此行为学表现定义为严重痫样发作(severe seizure,SS)。腹腔注射330 mg/kg和360 mg/kg匹罗卡品后,在癫痫急性期出现过SS的C57BL/6J小鼠,经过潜伏期后可出现SRS,C57BL/6J亚系小鼠造模后SRS的比例(70%)和经历过SE后出现SRS的C57BL/6N亚系小鼠(75%)相近。造模后28 d,C57BL/6J小鼠海马出现颞叶癫痫的特征性病理改变,包括苔藓纤维出芽和神经元丢失。结论C57BL/6J亚系小鼠经腹腔注射匹罗卡品诱导癫痫模型时,造模成功的行为学标准可以是SE的发生,也可以是SS的出现。Objective To explore the approach to establish a temporal lobe epilepsy model via intraperitoneal pilocarpine injection in C57BL/6J mice,and to summarize behavioral indicators predicting successful modeling during the acute phase of epileptic seizures after pilocarpine administration,aiming to offer a practical mice model for future epilepsy research.Methods Thirty C57BL/6J substrain mice(primary subjects)and forty C57BL/6N substrain mice(control subjects)were selected to establish a temporal lobe epilepsy model by inducing seizures through a single intraperitoneal injection of pilocarpine.The mice from the two substrains were each divided into 3 groups,and were injected intraperitoneally with 300 mg/kg,330 mg/kg,or 360 mg/kg of pilocarpine,respectively.Motor seizure behaviors were observed and compared between the two substrains of C57BL/6 mice post pilocarpine injection,and the spontaneous recurrent seizures(SRS)were continuously monitored from the 7th day after injection.On the 28th day post-injection,the mice were euthanized and the histopathological changes in their hippocampi were examined.Results After pilocarpine administration,C57BL/6N mice displayed characteristic motor seizures followed by the onset of status epilepticus(SE).Conversely,C57BL/6J mice showed fewer instances of typical motor seizure behavior and the subsequent SE.Instead,they more often exhibited systemic tremors lasting several seconds to tens of seconds following limb twitching.This behavior is classified as“severe seizure(SS)”in current study.Following intraperitoneal injection of 330 mg/kg and 360 mg/kg pilocarpine,C57BL/6J mice displaying SS during the acute phase of seizure might exhibit SRS after a latency period.The percentage of spontaneous seizures observed in C57BL/6J mice post-modeling(70%)was comparable to that seen in C57BL/6N mice(75%)which developed SRS subsequent to SE.C57BL/6J mice displayed characteristic pathological alterations associated with temporal lobe epilepsy in the hippocampi after 28 d following pilocarpi

关 键 词：小鼠模型 颞叶癫痫 匹罗卡品 C57BL/6J 苔藓纤维出芽 神经元丢失 

分 类 号：R742.1[医药卫生—神经病学与精神病学]