Page 60 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第43卷第8期
·1094 · Journal of Nanjing Medical University（Natural Sciences） 2023年8月

·基础研究·

梨状皮层注射红藻氨酸建立癫痫小鼠模型

张玲芝，秦 滢，吴连连，刘婷隽，陈全刚，胡安康 *
徐州医科大学实验动物中心，江苏 徐州 221004

［摘 要］ 目的：建立和评价梨状皮层微量注射红藻氨酸（kainic acid，KA）激发的癫痫小鼠模型。方法：利用即刻早期基因（c⁃
fos）免疫荧光染色技术、原位末端转移酶标记染色（TdT⁃mediated dUTP nick⁃end labeling，TUNEL）技术在急性癫痫小鼠模型中
确定癫痫相关脑区梨状皮层。在梨状皮层脑立体定位注射KA后，观察小鼠癫痫发作程度（Racine评分）和癫痫发作潜伏期，行
为学实验包括水迷宫检测小鼠学习记忆能力和新物体识别实验检测小鼠探索能力，癫痫发作结果和行为学实验结果均与海马
立体定位注射KA的癫痫模型作对比。利用电生理技术检测新模型小鼠神经元电活动。结果：c⁃fos免疫荧光和TUNEL染色结
果显示，在急性癫痫小鼠模型中除海马外，梨状皮层脑区神经元也被大量激活，且发生凋亡；在梨状皮层注射KA后成功激发
癫痫，与海马注射KA的癫痫模型相比，癫痫发作评分没有显著性差异且发作潜伏期更短；行为学结果显示，梨状皮层注射KA
的癫痫模型小鼠空间记忆能力降低，新物体探索能力降低，该结果与海马注射KA的癫痫模型相比无显著性差异；膜片钳实验
显示梨状皮层注射KA的癫痫模型神经元放电频率增加、幅值降低、静息膜电位上升。结论：在梨状皮层微量注射KA后成功
激发癫痫，这为癫痫发病机制的研究提供了一种新的动物模型。
［关键词］ 梨状皮层；海马；癫痫；红藻氨酸；c⁃fos
［中图分类号］ R742.1 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）08⁃1094⁃08
doi：10.7655/NYDXBNS20230808

Establishing a mouse model of epilepsy via injectin of kainic acid into the pirform contex
*
ZHANG Lingzhi，QIN Ying，WU Lianlian，LIU Tingjun，CHEN Quangang，HU Ankang
Laboratory Animal Center，Xuzhou Medical University，Xuzhou 221004，China

［Abstract］ Objective：The current study aims to establish and evaluatee the kindling model of epilepsy induced by kainic acid（KA）
microinjection into piriform cortex. Methods：Using c⁃fos immunofluorescence staining and TdT⁃mediated dUTP nick⁃end labeling
（TUNEL）staining techniques to find the piriform cortex associated with epilepsy in a acute epilepsy mouse model. Racine score and
seizure latency were observed after injection of KA in piriform cortex. Behavioral experiments were performed including testing the
learning and memory abilities of mice in a water maze and the exploration ability of mice in a new object. The results of seizures and
behavioral experiments were compared with the epileptic model of stereotactic injection of KA into the hippocampus. The electrical
activities of new model mouse neurons were measured by electrophysiological techniques. Results：The results of c ⁃ fos
immunofluorescence and TUNEL staining showed that in addition to the hippocampus，neurons in the piriform cortex and hippocampus
were also significantly activated，and apoptosis occurred in the acute epilepsy mouse model. After injecting KA into the piriform cortex，
the epilepsy was successfully ignited. Compared with the epilepsy model injected KA into the hippocampus，there was no significant
difference in the epileptic seizure score and the seizurelatency was shorter. The behavioral results showed that the epileptic model mice
induced by KA injection into the piriform cortex had a reduced spatial memory ability and a reduced ability to explore new objects.
These results showed no significant difference compared to the epileptic model induced by KA injection into the hippocampus. The
patch clamp recording test showed an increase in firing frequency and the resting membrane potential，and a decrease in amplitude of
neuronal discharge. Conclusion：The microinjection of KA into the piriform cortex has successfully induced epilepsy，which may
provide a new animal model for studying the pathogenesis of epilepsy.
［Key words］ piriform cortex；hippocampus；epilepsy；kainic acid；c⁃fos
［J Nanjing Med Univ，2023，43（08）：1094⁃1101］
［基金项目］ 江苏省自然科学基金（BK20170251）；江苏省高等学校自然科学基金（21KJB180011）；徐州市科技局社会发展项
目（ZYSB20210410）；江苏省实验动物协会科研基金（DWXH202206）；徐州市科技创新重点研发计划⁃面上项目（KC22078）
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通信作者（Corresponding author），E⁃mail：300112110963@stu.xzhmu.cn