戊四氮致癫大鼠不同脑区高迁移率族蛋白-1的表达与意义

发布时间：2018-04-20 18:40

本文选题：癫痫 + 戊四氮　；参考：《福建医科大学》2014年硕士论文

【摘要】：目的: 采用戊四氮(PTZ)诱发大鼠慢性癫痫，动态观察不同脑区高迁移率族蛋白-1(High-mobility group box-1,HMGB-1)含量的变化，以及探讨HMGB-1与癫痫的关系。方法: 1、慢性癫痫大鼠模型的建立 SD雄性大鼠32只，将其随机分为对照组（A组，n=8）和实验组（n=24），对照组予腹腔注射40mg·kg-1的生理盐水，实验组分别腹腔注射不同浓度的PTZ,其中B组（20mg·kg-1）、C组（40mg·kg-1）、D组（60mg·kg-1），每次注射后密切观察大鼠行为变化至少1小时，记录下大鼠点燃所需天数、潜伏期时间、点燃的数量、存活的数量。 2、PTZ致癫大鼠不同脑区HMGB-1的表达 SD雄性大鼠40只,将其随机分为正常对照组(A组，n=8)和实验组（n=32）。实验组大鼠采用PTZ40mg·kg-1一次性腹腔注射，对照组大鼠给予等量的生理盐水腹腔注射，根据癫痫发作后的时间，于其发作后1h（B组）、6h（C组）、24h (D组)、72h（E组）取脑，采用免疫组织化学方法，动态观察海马组织中HMGB-1的变化；RT-PCR方法动态观察脑组织中额叶、海马、中脑中HMGB-1的变化。结果： 1、当PTZ浓度为20mg·kg-1时，发作程度低，不易点燃；当PTZ浓度为60mg·kg-1时，发作较为剧烈，易点燃，但发作进展快，死亡率比较高，大鼠存活率低；当PTZ浓度为40mg·kg-1时，发作程度逐渐加重，点燃率较高，大鼠存活率高，不易死亡，发作缓解后仍可自由活动； 2、荧光定量PCR显示：幼鼠不同脑区（额叶、中脑、海马）HMGB-1蛋白表达随着时间的推移呈动态升高，其中额叶、海马于6h开始升高（P＜0.05），24h达高峰（P＜0.05），与24h相比，，72h有所降低（P＜0.05），但仍高于对照组（P＜0.05），中脑1h、6h、24h与对照组比较差异无统计学意义（P0.05），72小时的HMGB-1蛋白表达明显高于对照组，差异有显著性（P0.05）； 3、海马免疫组化结果显示，各组HMGB-1蛋白平均光密度值与对照组相比逐步升高，其中，B组HMGB-1蛋白表达量与对照组相比有所升高，但差异无统计学意义（P0.05）；与对照组相比，C组、D组HMGB-1蛋白表达量逐渐增高，差异有统计学意义（P0.05）；E组蛋白表达量有减少趋势，与D组相比，差异有统计学意义（P＜0.05），但亦显著高于对照组，差异有统计学意义（P0.01）。结论： 1、戊四氮腹腔注射法建立慢性癫痫动物模型的理想浓度为40mg·kg-1； 2、EP发作后HMGB-1蛋白表达明显升高，HMGB-1与癫痫所致大脑损伤密切相关。
[Abstract]:Objective: Chronic epilepsy was induced by pentylenetetrazol (PTZ) in rats. The changes of high mobility group box-1 HMGB-1) in different brain regions were observed dynamically, and the relationship between HMGB-1 and epilepsy was investigated. Methods: 1. Establishment of chronic epilepsy rat model Thirty-two male SD rats were randomly divided into two groups: control group (n = 8) and experimental group (n = 24). The control group was intraperitoneally injected with normal saline of 40mg kg-1. The experimental group was intraperitoneally injected with different concentrations of PTZs, including group B (20 mg / kg) and group C (40 mg / kg ~ (-1)) and group D (60 mg / kg ~ (-1)). The behavioral changes of rats were closely observed for at least one hour after each injection, and the days required for kindling, incubation period, the number of kindling and the number of surviving rats were recorded. Expression of HMGB-1 in different brain regions of epileptic rats induced by PTZ Forty Sprague-Dawley male rats were randomly divided into normal control group (group A) and experimental group (n = 32). The rats in the experimental group were injected intraperitoneally with PTZ40mg kg-1 and the rats in the control group were given the same amount of normal saline intraperitoneally. According to the time after seizure, the brain was taken from the rats in the 1h(B group at 6 h after seizure and in the D group at 24 h after seizure, and the rats in the control group were given the same amount of normal saline intraperitoneal injection. The changes of HMGB-1 in hippocampus were observed dynamically. The changes of HMGB-1 in frontal lobe, hippocampus and midbrain were observed dynamically by RT-PCR. Results: 1. When the concentration of PTZ was 20mg kg-1, the attack degree was low and it was not easy to ignite. When PTZ concentration was 60mg kg-1, the attack was more intense and easy to ignite, but the attack progression was faster, the mortality was higher, and the survival rate of rats was low. When the PTZ concentration was 40mg kg-1, the attack degree gradually aggravated. The rate of kindling was high, the survival rate of rats was high, and it was not easy to die. 2, fluorescence quantitative PCR showed that the expression of HMGB-1 protein in different brain regions (frontal lobe, midbrain, hippocampus) of young rats increased with the passage of time. The hippocampus began to increase at 6 h, P < 0.05 and reached the peak at 24 h (P < 0.05). Compared with the control group, the expression of HMGB-1 protein at 72h was lower than that in the control group (P < 0.05), but still higher than that in the control group (P < 0.05). There was no significant difference in the expression of HMGB-1 protein between the control group and the control group at 1h ~ 6h, and the expression of HMGB-1 protein was significantly higher than that of the control group at 72h (P 0.05). 3. The immunohistochemical results of hippocampus showed that the average optical density of HMGB-1 protein in each group was higher than that in control group, and the expression of HMGB-1 protein in group B was higher than that in control group. Compared with the control group, the expression of HMGB-1 protein in the C group increased gradually, and the difference was significant (P < 0.05), but it was also significantly higher than that in the control group (P < 0.05), the difference was significant (P < 0.05), and the difference was significant (P < 0.05), but it was also significantly higher than that in the control group (P < 0.05), and the difference was significant (P < 0.05), and the difference was significant (P < 0.05). The difference was statistically significant (P 0.01). Conclusion: 1. The ideal concentration of pentylenetetrazol in the establishment of chronic epileptic animal model was 40mg kg-1; 2the expression of HMGB-1 protein increased significantly after EP attack. HMGB-1 was closely related to brain injury induced by epilepsy.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742.1

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