【摘要】：目的：通過(guò)氯化鋰-匹羅卡品小劑量反復(fù)腹腔注射誘導(dǎo)昆明小鼠癲癇持續(xù)狀態(tài)發(fā)作,模擬人類顳葉癲癇動(dòng)物模型,分析DYRK1A mRNA在小鼠腦組織中的表達(dá)情況及其與海馬硬化形成的相關(guān)性,探討DYRK1A是否參與難治性顳葉癲癇的形成,為繼續(xù)研究DYRK1A蛋白與海馬硬化發(fā)生的相關(guān)性提供理論依據(jù)。 方法：將清潔級(jí)健康雄性昆明小鼠50只,隨機(jī)分為正常對(duì)照組16只和致癇組34只,分別予以生理鹽水和氯化鋰-匹羅卡品小劑量反復(fù)腹腔注射,建立小鼠癲癇持續(xù)狀態(tài)(SE)；分別觀察24小時(shí)和30天,隨機(jī)選取6只為急性致癲組,慢性致癲組6只,通過(guò)行為學(xué)、電生理學(xué)、病理學(xué)HE染色方法驗(yàn)證模型的可靠性,建立人類顳葉癲癇小鼠模型。采用逆轉(zhuǎn)錄PCR方法檢測(cè)DYRK1A mRNA在小鼠腦組織中的表達(dá)水平。 結(jié)果：氯化鋰-匹羅卡品小劑量反復(fù)腹腔注射昆明小鼠共34只,其中有22只小鼠誘發(fā)癲癇持續(xù)狀態(tài)(SE),發(fā)作達(dá)到Racine標(biāo)準(zhǔn)Ⅳ-Ⅴ級(jí),剔除12只發(fā)作未達(dá)到Racine標(biāo)準(zhǔn)Ⅳ-Ⅴ級(jí)者。進(jìn)入SE的22只小鼠中24小時(shí)內(nèi)死亡的有5只,SE后24小時(shí)隨機(jī)選取6只入選急性致癇組；24-72小時(shí)后死亡的有3只。其余8只小鼠在24-72小時(shí)后進(jìn)入靜止期,觀察30天,隨機(jī)選取6只進(jìn)入慢性致癲組。6只正常對(duì)照組小鼠,腹腔注射生理鹽水后未見(jiàn)發(fā)作,未見(jiàn)死亡。本研究中,SE誘發(fā)成功率為64.71%(22/34),SE后死亡率為36.36%(8/22),；自發(fā)癲癇發(fā)作出現(xiàn)率為47.06%(8/17)。病理學(xué)HE染色分析,與正常對(duì)照組相比,致癲組海馬神經(jīng)元細(xì)胞排列不整齊,細(xì)胞間隙增大,出現(xiàn)腫脹、變性、壞死、崩解,胞體固縮,體積變小,胞漿濃縮深染,細(xì)胞核固縮,核仁不清,死亡細(xì)胞胞核裂解,細(xì)胞漿消失。致癲組小鼠模型的腦電監(jiān)測(cè)可見(jiàn)：急性期爆發(fā)長(zhǎng)程出現(xiàn)的尖活動(dòng)、棘活動(dòng)以及不規(guī)則的尖慢復(fù)合活動(dòng),明顯突出于背景腦電活動(dòng)；慢性期散在出現(xiàn)的、頻率慢于背景活動(dòng)的慢活動(dòng)以及癇性放電。6例正常對(duì)照組DYRK1A mRNA與β-actin比值為0.4830±0.1243；6例急性致癲組DYRK1A mRNA與β-actin比值為0.8883±0.0727；6例慢性致癲組DYRKlAmRNA與β-actin比值為0.7112±0.1216；三組中兩兩比較差異均有顯著性統(tǒng)計(jì)學(xué)意義(P0.05)。DYRKIA mRNA在三組中的表達(dá)情況：急性致癲組慢性致癲組正常對(duì)照組。 結(jié)論：氯化鋰-匹羅卡品誘導(dǎo)昆明小鼠癲癇持續(xù)狀態(tài)后,腦組織中DYRKIA表達(dá)量較正常對(duì)照組增加。
[Abstract]:Objective: to induce the seizure of status epilepticus in Kunming mice by repeated intraperitoneal injection of lithium chloride-pilocarpine at low dose, and to simulate the animal model of temporal lobe epilepsy in humans. To analyze the expression of DYRK1A mRNA in the brain of mice and its correlation with hippocampal sclerosis, to explore whether DYRK1A is involved in the formation of refractory temporal lobe epilepsy, and to provide theoretical basis for further study on the correlation between DYRK1A protein and hippocampal sclerosis. Methods: fifty healthy Kunming mice of clean grade were randomly divided into normal control group (n = 16) and epilepsy group (n = 34). Normal saline and lithium chloride-pilocarpine were repeatedly injected intraperitoneally in order to establish status epilepticus status (SE);) in mice. After 24 hours and 30 days of observation, 6 mice were randomly selected as acute epilepsy group and 6 chronic epilepsy group. The reliability of the model was verified by behavioral, electrophysiological and pathological HE staining methods, and the human temporal lobe epilepsy mice model was established. Reverse transcription-PCR (RT-PCR) was used to detect the expression of DYRK1A mRNA in mouse brain tissue. Results: 34 Kunming mice were repeatedly injected with lithium chloride-pilocarpine intraperitoneally. Among them, 22 mice induced status epilepticus (SE), attack to Racine grade 鈪，

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