DYRK1A表达与难治性颞叶癫痫的相关性研究
发布时间：2019-03-12 12:27
【摘要】：目的：通过氯化锂-匹罗卡品小剂量反复腹腔注射诱导昆明小鼠癫痫持续状态发作,模拟人类颞叶癫痫动物模型,分析DYRK1A mRNA在小鼠脑组织中的表达情况及其与海马硬化形成的相关性,探讨DYRK1A是否参与难治性颞叶癫痫的形成,为继续研究DYRK1A蛋白与海马硬化发生的相关性提供理论依据。 方法：将清洁级健康雄性昆明小鼠50只,随机分为正常对照组16只和致痫组34只,分别予以生理盐水和氯化锂-匹罗卡品小剂量反复腹腔注射,建立小鼠癫痫持续状态(SE)；分别观察24小时和30天,随机选取6只为急性致癫组,慢性致癫组6只,通过行为学、电生理学、病理学HE染色方法验证模型的可靠性,建立人类颞叶癫痫小鼠模型。采用逆转录PCR方法检测DYRK1A mRNA在小鼠脑组织中的表达水平。 结果：氯化锂-匹罗卡品小剂量反复腹腔注射昆明小鼠共34只,其中有22只小鼠诱发癫痫持续状态(SE),发作达到Racine标准Ⅳ-Ⅴ级,剔除12只发作未达到Racine标准Ⅳ-Ⅴ级者。进入SE的22只小鼠中24小时内死亡的有5只,SE后24小时随机选取6只入选急性致痫组；24-72小时后死亡的有3只。其余8只小鼠在24-72小时后进入静止期,观察30天,随机选取6只进入慢性致癫组。6只正常对照组小鼠,腹腔注射生理盐水后未见发作,未见死亡。本研究中,SE诱发成功率为64.71%(22/34),SE后死亡率为36.36%(8/22),；自发癫痫发作出现率为47.06%(8/17)。病理学HE染色分析,与正常对照组相比,致癫组海马神经元细胞排列不整齐,细胞间隙增大,出现肿胀、变性、坏死、崩解,胞体固缩,体积变小,胞浆浓缩深染,细胞核固缩,核仁不清,死亡细胞胞核裂解,细胞浆消失。致癫组小鼠模型的脑电监测可见：急性期爆发长程出现的尖活动、棘活动以及不规则的尖慢复合活动,明显突出于背景脑电活动；慢性期散在出现的、频率慢于背景活动的慢活动以及痫性放电。6例正常对照组DYRK1A mRNA与β-actin比值为0.4830±0.1243；6例急性致癫组DYRK1A mRNA与β-actin比值为0.8883±0.0727；6例慢性致癫组DYRKlAmRNA与β-actin比值为0.7112±0.1216；三组中两两比较差异均有显著性统计学意义(P0.05)。DYRKIA mRNA在三组中的表达情况：急性致癫组慢性致癫组正常对照组。 结论：氯化锂-匹罗卡品诱导昆明小鼠癫痫持续状态后,脑组织中DYRKIA表达量较正常对照组增加。
[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，
本文编号：2438773
[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，
本文编号：2438773
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