KLF4在PTZ点燃模型大鼠海马苔藓纤维出芽过程中的动态表达
发布时间：2019-01-02 17:17
【摘要】：目的采用戊四氮(Pentamethazol, PTZ)点燃癫痫大鼠模型,观察海马中转录因子Kruppe样因子4(Kruppel-like factor4, KLF4)的表达变化,结合海马苔藓纤维出芽(Mossy fiber sprouting, MFS)的动态改变,探讨其在癫痫发生过程中的可能作用。 方法清洁级健康成年雄性SD大鼠90只,随机分为戊四氮实验组和生理盐水对照组,其中戊四氮实验组60只,生理盐水对照组30只。戊四氮组腹腔注射PTZ30mg/kg,每天一次,对照组同时注射等量生理盐水。按第一次腹腔注射后3天、1周、2周、4周和6周在两组内随机平均分为5个亚组,每个亚组再随机平均分为两个小组：一组用于免疫组化,检测各时间点大鼠海马中的KLF4表达；另一组用于Timm染色,检测苔藓纤维出芽情况。 结果 1.动物模型 大鼠多在连续注射PTZ21～28天后被点燃,平均24.5±2.3天。模型点燃率为100%,死亡率为2.2%。实验组大鼠成功点燃后出现诱发发作及自发发作；对照组未出现任何痫性发作。 2. KLF4蛋白的表达 对照组大鼠海马CA1、CA3、齿状回区KLF4蛋白表达在3天后逐渐升高,至1周时达高峰,并持续至第6周；实验组大鼠海马CA1、CA3、齿状回区KLF4蛋白表达在第3天后未升高,而是保持在稳定水平,持续至第6周。两组间CA1区KLF4蛋白表达在3天、1周时有统计学差异；CA3区仅1周组有统计学差异；齿状回区在1周和2周组有统计学差异；余各区各时间点两组间无统计学差异。 3.苔藓纤维出芽的动态变化 实验组大鼠海马CA3区苔藓纤维出芽与对照组CA3区相比,从第3天开始实验组出现苔藓纤维出芽,评分逐渐增加,至第2周开始两组间出现统计学差异(P0.05),至第6周时达高峰。实验组大鼠海马齿状回区苔藓纤维出芽与对照组齿状回区相比,从第4周开始实验组出现苔藓纤维出芽,评分逐渐增加,至第6周时达高峰并在两组间出现统计学差异(P0.05)。对照组大鼠海马CA3区苔藓纤维出芽评分均仅0-1分,各时间点间无统计学差异；齿状回区各时间点未发现苔藓纤维出芽。 结论 在戊四氮点燃大鼠模型中,海马KLF4蛋白早期表达下调可能促进苔藓纤维出芽。
[Abstract]:Objective to observe the expression of transcription factor Kruppe like factor 4 (Kruppel-like factor4, KLF4) in hippocampus of epileptic rats kindled with pentylenetetrazol (Pentamethazol, PTZ), and to observe the dynamic changes of (Mossy fiber sprouting, MFS) in mossy fibers. To explore the possible role of Epilepsy in the process of epilepsy. Methods 90 healthy male SD rats of clean grade were randomly divided into pentylenetetrazol group (n = 60) and saline control group (n = 30). Pentylenetetrazol group was intraperitoneally injected with PTZ30mg/kg, once a day, while the control group was injected with the same amount of normal saline at the same time. Three days, one week, two weeks, four weeks and six weeks after the first intraperitoneal injection, each subgroup was randomly divided into five subgroups, and each subgroup was randomly divided into two groups: one for immunohistochemistry. The expression of KLF4 in hippocampus of rats was detected at different time points. The other group was used for Timm staining to detect the sprouting of moss fibers. Result 1. Most of the animal model rats were kindled after continuous injection of PTZ21~28, with an average of 24.5 卤2.3 days. The model ignition rate was 100 and the mortality was 2.2. The rats in the experimental group had induced seizures and spontaneous seizures after successful kindling, while in the control group there were no epileptic seizures. 2. The expression of KLF4 protein in hippocampal CA1,CA3, dentate gyrus of control group increased gradually after 3 days, reached its peak at 1 week and lasted until 6 weeks. The expression of KLF4 protein in the dentate gyrus of hippocampal CA1,CA3, did not increase after 3 days in the experimental group, but remained at a stable level until 6 weeks. The expression of KLF4 protein in CA1 region was significantly different between the two groups at 3 days and 1 week; in the CA3 region only 1 week group there was statistical difference; in dentate gyrus area at 1 week and 2 weeks group there was statistical difference; there was no statistical difference between the two groups at other time points. 3. Dynamic changes of mossy fiber sprouting in the experimental group compared with the control group, the mossy fiber sprouts appeared in the experimental group from the 3rd day, and the score increased gradually compared with the control group. At the beginning of the second week, there was a statistical difference between the two groups (P0.05), and reached the peak at the 6th week. Compared with the control group, the mossy fiber sprouts from the dentate gyrus of the experimental group and the control group, the mossy fiber sprouts appeared in the experimental group from the 4th week, and the score increased gradually. Reached the peak at the 6th week and there was statistical difference between the two groups (P0.05). In the control group, the mossy fiber sprouting scores in the hippocampal CA3 area were only 0-1, and there was no statistical difference between the different time points, and no mossy fiber sprouting was found in the dentate gyrus at each time point. Conclusion in pentylenetetrazol kindled rat model, early down-regulation of hippocampal KLF4 protein may promote mossy fiber sprouting.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R742.1;R-332
本文编号：2398764
[Abstract]:Objective to observe the expression of transcription factor Kruppe like factor 4 (Kruppel-like factor4, KLF4) in hippocampus of epileptic rats kindled with pentylenetetrazol (Pentamethazol, PTZ), and to observe the dynamic changes of (Mossy fiber sprouting, MFS) in mossy fibers. To explore the possible role of Epilepsy in the process of epilepsy. Methods 90 healthy male SD rats of clean grade were randomly divided into pentylenetetrazol group (n = 60) and saline control group (n = 30). Pentylenetetrazol group was intraperitoneally injected with PTZ30mg/kg, once a day, while the control group was injected with the same amount of normal saline at the same time. Three days, one week, two weeks, four weeks and six weeks after the first intraperitoneal injection, each subgroup was randomly divided into five subgroups, and each subgroup was randomly divided into two groups: one for immunohistochemistry. The expression of KLF4 in hippocampus of rats was detected at different time points. The other group was used for Timm staining to detect the sprouting of moss fibers. Result 1. Most of the animal model rats were kindled after continuous injection of PTZ21~28, with an average of 24.5 卤2.3 days. The model ignition rate was 100 and the mortality was 2.2. The rats in the experimental group had induced seizures and spontaneous seizures after successful kindling, while in the control group there were no epileptic seizures. 2. The expression of KLF4 protein in hippocampal CA1,CA3, dentate gyrus of control group increased gradually after 3 days, reached its peak at 1 week and lasted until 6 weeks. The expression of KLF4 protein in the dentate gyrus of hippocampal CA1,CA3, did not increase after 3 days in the experimental group, but remained at a stable level until 6 weeks. The expression of KLF4 protein in CA1 region was significantly different between the two groups at 3 days and 1 week; in the CA3 region only 1 week group there was statistical difference; in dentate gyrus area at 1 week and 2 weeks group there was statistical difference; there was no statistical difference between the two groups at other time points. 3. Dynamic changes of mossy fiber sprouting in the experimental group compared with the control group, the mossy fiber sprouts appeared in the experimental group from the 3rd day, and the score increased gradually compared with the control group. At the beginning of the second week, there was a statistical difference between the two groups (P0.05), and reached the peak at the 6th week. Compared with the control group, the mossy fiber sprouts from the dentate gyrus of the experimental group and the control group, the mossy fiber sprouts appeared in the experimental group from the 4th week, and the score increased gradually. Reached the peak at the 6th week and there was statistical difference between the two groups (P0.05). In the control group, the mossy fiber sprouting scores in the hippocampal CA3 area were only 0-1, and there was no statistical difference between the different time points, and no mossy fiber sprouting was found in the dentate gyrus at each time point. Conclusion in pentylenetetrazol kindled rat model, early down-regulation of hippocampal KLF4 protein may promote mossy fiber sprouting.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R742.1;R-332
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