PTZ点燃大鼠海马FAKpY397动态表达及其与苔藓纤维出芽关系
发布时间：2018-12-14 11:17
【摘要】：目的通过观察戊四氮点燃大鼠模型海马FAKpY397蛋白的动态表达变化,结合苔藓纤维出芽的程度,探讨其在苔藓纤维出芽及癫痫致痫过程中的作用。 方法将135只健康的成年雄性Sprague-Dawley (SD)大鼠,随机分为实验组90只和对照组45只。实验组每天腹腔注射亚惊厥剂量的PTZ(即戊四氮—35mg/kg,对照组每天腹腔注射同等量生理盐水,成功点燃后停止注射(痫性发作等级评定标准参照Racine标准)。根据首次腹腔注射后3天,1周,2周,4周和6周将实验组及对照组分别随机分为五个亚组。根据不同的实验方法及用途再将每个不同的再亚组随机细分为三个小组：一组用于免疫荧光及免疫组化染色,检测各不同时间点实验组与对照组FAKpY397蛋白在海马的表达分布及表达量的变化；一组用于Western Blot,检测不同时间点不同组别大鼠海马区FAKpY397蛋白总表达量及其变化趋势；另一组用于Timm染色,检测海马苔藓纤维出芽情况并进行评分。其中实验组每小组各6只,对照组每小组各3只。 结果 1.动物行为学 随着每天相同亚惊厥剂量PTZ经腹腔注射给药,实验组大鼠所表现的痫性发作反应逐渐加重,多数给药后第21-28天(23.1±-2.3天)被成功点燃。PTZ致痫模型点燃率为93.75%,而死亡率为4.44%。被成功点燃的大鼠可见Racine I至RacineⅢ级的自发痫性发作。对照组大鼠未出现痫性发作,行为正常。 2.Western Blot结果显示：在PTZ点燃致痫过程中,海马FAKpY397蛋白总表达量于再PTZ腹腔注射后3d稍下降,1周时开始增加,4周达高峰,6周下降,仍高于对照组。除3天组外(0.05),实验组各时间点与对照组间比较均有统计学差异(P0.05)。 3.免疫荧光及组化结果显示：FAKpY397在海马各区有表达,主要分布于胞浆；免疫组化半定量分析示CA3区FAKpY397蛋白表达于PTZ腹腔注射后3d开始增加,4周达高峰,6周下降,仍高于对照组。实验组中除1周组与2周组进行比较无统计学差异(P0.05),其余各时间点两两比较及实验组与对照组对应时间点比较均有统计学差异(P0.05)。 4.苔藓纤维出芽评分 对照组CA3区未见苔藓纤维出芽或仅见轻微出芽,苔藓纤维出芽评分在各时间点间两两比较均无统计学差异。实验组CA3区苔藓纤维出芽评分从PTZ腹腔注射后3天开始增加,4周时达高峰,6周下降。实验组中除4周与6周间比较无统计学差异(0.05),其余各时间点两两比较及实验组与对照组对应时间点比较均有统计学差异(P0.05)。 结论 FAKpY397蛋白在海马CA3区表达增多可能对于苔藓纤维出芽具有促进作用进而诱发癫痫发作。
[Abstract]:Objective to investigate the role of pentylenetetrazol (PTZ) in the process of mossy fiber sprouting and epilepsy by observing the dynamic expression of FAKpY397 protein in hippocampus of rats with pentylenetetrazol (PTZ) kindling and combining with the degree of mossy fiber sprouting. Methods 135 healthy male Sprague-Dawley (SD) rats were randomly divided into experimental group (n = 90) and control group (n = 45). Subconvulsant dose of PTZ (pentylenetetrazo-35 mg / kg) was injected intraperitoneally every day in the experimental group, and the control group was injected with the same amount of normal saline daily. The injection was stopped after successful kindling (Racine criterion was used to evaluate the grade of epileptic seizures). According to 3 days, 1 week, 2 weeks, 4 weeks and 6 weeks after the first intraperitoneal injection, the experimental group and the control group were randomly divided into five subgroups. Each subgroup was subdivided randomly into three groups according to different experimental methods and uses: one group was used for immunofluorescence and immunohistochemical staining. The expression distribution and quantity of FAKpY397 protein in hippocampus of experimental group and control group were detected at different time points. One group was used to detect the total expression of FAKpY397 protein and its changing trend at different time points in different groups of rats, the other group was used for Timm staining to detect and score the budding of mossy fibers in hippocampus. There were 6 rats in each group in the experimental group and 3 rats in the control group. Result 1. With the same daily subconvulsive dose of PTZ administered intraperitoneally, the epileptic seizure response in the experimental group increased gradually. Most of the drugs were ignited 21-28 days (23.1 卤-2.3 days) after administration. The kindling rate of epileptic model induced by PTZ was 93.755.The death rate was 4.44g. Spontaneous epileptic seizures of Racine I to Racine 鈪，
本文编号：2378524
[Abstract]:Objective to investigate the role of pentylenetetrazol (PTZ) in the process of mossy fiber sprouting and epilepsy by observing the dynamic expression of FAKpY397 protein in hippocampus of rats with pentylenetetrazol (PTZ) kindling and combining with the degree of mossy fiber sprouting. Methods 135 healthy male Sprague-Dawley (SD) rats were randomly divided into experimental group (n = 90) and control group (n = 45). Subconvulsant dose of PTZ (pentylenetetrazo-35 mg / kg) was injected intraperitoneally every day in the experimental group, and the control group was injected with the same amount of normal saline daily. The injection was stopped after successful kindling (Racine criterion was used to evaluate the grade of epileptic seizures). According to 3 days, 1 week, 2 weeks, 4 weeks and 6 weeks after the first intraperitoneal injection, the experimental group and the control group were randomly divided into five subgroups. Each subgroup was subdivided randomly into three groups according to different experimental methods and uses: one group was used for immunofluorescence and immunohistochemical staining. The expression distribution and quantity of FAKpY397 protein in hippocampus of experimental group and control group were detected at different time points. One group was used to detect the total expression of FAKpY397 protein and its changing trend at different time points in different groups of rats, the other group was used for Timm staining to detect and score the budding of mossy fibers in hippocampus. There were 6 rats in each group in the experimental group and 3 rats in the control group. Result 1. With the same daily subconvulsive dose of PTZ administered intraperitoneally, the epileptic seizure response in the experimental group increased gradually. Most of the drugs were ignited 21-28 days (23.1 卤-2.3 days) after administration. The kindling rate of epileptic model induced by PTZ was 93.755.The death rate was 4.44g. Spontaneous epileptic seizures of Racine I to Racine 鈪，
本文编号：2378524
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