细胞因子SDF-1、RANTES、IL-2和IL-6在癫痫初期的表达研究
摘要
[目的]通过研究细胞因子SDF-1、RANTES、L-2、IL-6在癫痫初期(SE 30min,SE 1hr,SE 2hr,SE 1day)的表达情况,来探讨这四个基因是否参与了癫痫持续状态初期的病理变化,以便更好地了解癫痫的发生发展过程,为治疗癫痫奠定基础。
[方法]选用4-6周的体重为18-20g的KM小鼠,建立匹罗卡品癫痫持续状态模型,将其随机分为5组即(1)对照组(注射生理盐水组)、(2)SE 30min组(注射匹罗卡品诱导癫痫持续状态30min组)、(3)SE lhr组(注射匹罗卡品诱导癫痫持续状态lhr组)、(4)SE 2hr组(注射匹罗卡品诱导癫痫持续状态2hr组)、(5)SE 1day组(注射匹罗卡品诱导癫痫持续状态后24hr组,其中癫痫持续状态时间大于2hr)。灌注固定后取大脑制作石蜡切片,采用免疫组织化学方法检测细胞因子SDF-1、RANTES、IL-2和IL-6在海马区域的表达变化。使用SPSS11.5软件包对数据进行统计学处理。
[结果]1.动物行为学观察,实验组小鼠腹腔注射匹罗卡品后,均出现Ⅳ级以上癫痫发作；对照组小鼠腹腔注射等量生理盐水,未见上述痫性发作。2.在光镜下观察,SDF-1、RANTES、IL-2、IL-6的阳性反应主要分布于海马CA1-CA4区的锥体细胞及齿状回区域的颗粒细胞的中间神经元中。3.免疫组化染色显示,在对照组小鼠海马组织中,SDF-1、RANTES、IL-2、IL-6均呈低水平的表达；4.与对照组小鼠相比,SE 30min、SE lhr、SE 2hr和SE 1day时SDF-1、RANTES、IL-2和IL-6在小鼠海马中的表达均上调；其中SE 30min、SE 1hr到SE 2hr的表达是逐渐增强的趋势,而到了SE 1day的时候,它们的表达又出现下降,比SE 2hr较低,但仍高于对照组,差异均具有显著性意义(P<0.01)。
[结论]1.在光镜下观察,SDF-1、RANTES、IL-2、IL-6阳性反应主要分布于海马CA1-CA4区的锥体细胞及齿状回区域的颗粒细胞的中间神经元中。2.在对照组小鼠海马组织中SDF-1、RANTES、IL-2和工L-6的低水平表达.显示它们可能参与了海马的正常生理过程。3.SDF-1、RANTES、IL-2和工L-6在癫痫初期的表达上调,揭示这四个基因参与了癫痫持续状态初期的病理过程,与癫痫的发生发展有关。
[Objective]Cytokines SDF-1、RANTES, IL-2 and IL-6 expression in the early epilepsy(SE 30min, SE 1hr,SE 2hr,SE 1day),it used to explore whether the four genes involved in the pathological changes of the early status epilepticus,so that we can better understand the occurrence and development of epilepsy, and better treat it.
[Methods]We used 18-20g KM mice which is 4-6 weeks to establish the model of status epilepticus, it was randomly divided into 5 groups, namely(1) control group (injected with saline group)、(2) SE 30min group (30min during pilocarpine-induced status epilepticus group)、(3) SE 1hr group (lhr during pilocarpine-induced status epilepticus group)、(4) SE 2hr group(2hr during pilocarpine-induced status epilepticus groups (5) SE 1day group (24hr after pilocarpine-induced status epilepticus group,its time is longer than 2hr). we made the brain paraffin sections, which is used perfusion to fixed the tissue before doing the brain histology, and used immunohistochemistry to detect cytokines SDF-1、RANTES、IL-2 and IL-6 expression in the hippocampus. We used SPSS 11.5 software package for statistical analysis.
[Results] 1. In observation of animal behavior, there were more than gradeⅣseizures after experimental group mice intraperitoneal injection of pilocarpine; control mice injected with normal saline, no such seizures.2.In the light microscope, SDF-1、RANTES、IL-2、IL-6 immunopositive product was mainly localized in intemeurons of pyramidal cells in CA1-4 areas and granule cells in the dentate gyrus of the hippocampus.3. Immunohistochemical staining showed that,in control mice hippocampus, SDF-1、RANTES、IL-2、IL-6 showed a low levels of expression. 4.Compared with the control group mice, the SDF-1、RANTES、IL-2 and IL-6 which is SE 30min、SE 1hr、SE 2hr and SE lday expression that occurs in mouse hippocampus are increased; its expression which occurs in SE 30mn、SE 1hr to SE 2hr is gradually increase, and it decline in SE 1day, which is lower than SE 2hr, but still higher than the control group,the differences were significant. (P<0.01).
[Conclusion] 1.In the light microscope, SDF-1、RANTES、IL-2、IL-6 immunopositive product was mainly localized in interneurons of pyramidal cells in CA1-4 areas and granule cells in the dentate gyrus of the hippocampus. 2. In the control mice hippocampus, SDF-1、RANTES、IL-2 and IL-6 expression of low levels. It showed that they may involved in the normal physiological processes of hippocampus. 3. SDF-1、RANTES、IL-2 and IL-6 expression increased in the early epilepsy, it revealed that four gene involved in the pathological process of initial status epilepticus, and they related to the occurrence and development of epilepsy.
[方法]选用4-6周的体重为18-20g的KM小鼠,建立匹罗卡品癫痫持续状态模型,将其随机分为5组即(1)对照组(注射生理盐水组)、(2)SE 30min组(注射匹罗卡品诱导癫痫持续状态30min组)、(3)SE lhr组(注射匹罗卡品诱导癫痫持续状态lhr组)、(4)SE 2hr组(注射匹罗卡品诱导癫痫持续状态2hr组)、(5)SE 1day组(注射匹罗卡品诱导癫痫持续状态后24hr组,其中癫痫持续状态时间大于2hr)。灌注固定后取大脑制作石蜡切片,采用免疫组织化学方法检测细胞因子SDF-1、RANTES、IL-2和IL-6在海马区域的表达变化。使用SPSS11.5软件包对数据进行统计学处理。
[结果]1.动物行为学观察,实验组小鼠腹腔注射匹罗卡品后,均出现Ⅳ级以上癫痫发作；对照组小鼠腹腔注射等量生理盐水,未见上述痫性发作。2.在光镜下观察,SDF-1、RANTES、IL-2、IL-6的阳性反应主要分布于海马CA1-CA4区的锥体细胞及齿状回区域的颗粒细胞的中间神经元中。3.免疫组化染色显示,在对照组小鼠海马组织中,SDF-1、RANTES、IL-2、IL-6均呈低水平的表达；4.与对照组小鼠相比,SE 30min、SE lhr、SE 2hr和SE 1day时SDF-1、RANTES、IL-2和IL-6在小鼠海马中的表达均上调；其中SE 30min、SE 1hr到SE 2hr的表达是逐渐增强的趋势,而到了SE 1day的时候,它们的表达又出现下降,比SE 2hr较低,但仍高于对照组,差异均具有显著性意义(P<0.01)。
[结论]1.在光镜下观察,SDF-1、RANTES、IL-2、IL-6阳性反应主要分布于海马CA1-CA4区的锥体细胞及齿状回区域的颗粒细胞的中间神经元中。2.在对照组小鼠海马组织中SDF-1、RANTES、IL-2和工L-6的低水平表达.显示它们可能参与了海马的正常生理过程。3.SDF-1、RANTES、IL-2和工L-6在癫痫初期的表达上调,揭示这四个基因参与了癫痫持续状态初期的病理过程,与癫痫的发生发展有关。
[Objective]Cytokines SDF-1、RANTES, IL-2 and IL-6 expression in the early epilepsy(SE 30min, SE 1hr,SE 2hr,SE 1day),it used to explore whether the four genes involved in the pathological changes of the early status epilepticus,so that we can better understand the occurrence and development of epilepsy, and better treat it.
[Methods]We used 18-20g KM mice which is 4-6 weeks to establish the model of status epilepticus, it was randomly divided into 5 groups, namely(1) control group (injected with saline group)、(2) SE 30min group (30min during pilocarpine-induced status epilepticus group)、(3) SE 1hr group (lhr during pilocarpine-induced status epilepticus group)、(4) SE 2hr group(2hr during pilocarpine-induced status epilepticus groups (5) SE 1day group (24hr after pilocarpine-induced status epilepticus group,its time is longer than 2hr). we made the brain paraffin sections, which is used perfusion to fixed the tissue before doing the brain histology, and used immunohistochemistry to detect cytokines SDF-1、RANTES、IL-2 and IL-6 expression in the hippocampus. We used SPSS 11.5 software package for statistical analysis.
[Results] 1. In observation of animal behavior, there were more than gradeⅣseizures after experimental group mice intraperitoneal injection of pilocarpine; control mice injected with normal saline, no such seizures.2.In the light microscope, SDF-1、RANTES、IL-2、IL-6 immunopositive product was mainly localized in intemeurons of pyramidal cells in CA1-4 areas and granule cells in the dentate gyrus of the hippocampus.3. Immunohistochemical staining showed that,in control mice hippocampus, SDF-1、RANTES、IL-2、IL-6 showed a low levels of expression. 4.Compared with the control group mice, the SDF-1、RANTES、IL-2 and IL-6 which is SE 30min、SE 1hr、SE 2hr and SE lday expression that occurs in mouse hippocampus are increased; its expression which occurs in SE 30mn、SE 1hr to SE 2hr is gradually increase, and it decline in SE 1day, which is lower than SE 2hr, but still higher than the control group,the differences were significant. (P<0.01).
[Conclusion] 1.In the light microscope, SDF-1、RANTES、IL-2、IL-6 immunopositive product was mainly localized in interneurons of pyramidal cells in CA1-4 areas and granule cells in the dentate gyrus of the hippocampus. 2. In the control mice hippocampus, SDF-1、RANTES、IL-2 and IL-6 expression of low levels. It showed that they may involved in the normal physiological processes of hippocampus. 3. SDF-1、RANTES、IL-2 and IL-6 expression increased in the early epilepsy, it revealed that four gene involved in the pathological process of initial status epilepticus, and they related to the occurrence and development of epilepsy.
引文
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