【作者】 赵岚；

【导师】 王洪新；

【作者基本信息】 天津医科大学 ， 神经病学（专业学位）， 2014， 硕士

【摘要】 目的本研究通过建立糖耐量受损大鼠模型,检测不同时间点大鼠认知功能情况,并检测不同时间点大鼠脑组织炎性通路中关键因子的表达情况,分析大鼠认知功能与炎性因子表达的关系,以求进一步探讨糖耐量受损对大鼠认知功能的影响,及其相关机制,为指导临床工作提供理论依据。方法选取新生Wistar大鼠80只(体重180-220g)。随机分为实验组和对照组,每组大鼠40只,实验组为糖耐量受损模型组,用高脂高糖饲料喂养,实验动物及饲料由天津医科大学动物实验中心提供。5周后测定大鼠血糖情况,并分别于第5周,10周,15周,20周采用水迷宫方法评定大鼠认知情况,采用免疫组化和原位杂交技术测定大鼠脑组织核转录因子(factor-kappa B, NF-κB),白细胞介素-1β (interleukin-1β, IL-1β)水平。对照组,普通饲料喂养,分别于第5周,10周,15周,20周,进行认知功能评定和测定炎性因子表达水平。结果水迷宫实验：实验组大鼠在实验初期学习及记忆能力较对照组无显著差异,随着饲养时间的延长,实验组大鼠逐渐成模,于饲养第15周全部成模,之后反复监测大鼠认知功能提示实验组大鼠与对照组相比学习及记忆能力减退,于第15周后有显著差异(P<0.05)。HE染色：与对照组相比,实验组光镜下偶可见凋亡细胞,胞浆红染,细胞核染色质聚集成团块状,由于凋亡细胞迅速被吞噬,又无炎症反应,因此在常规切片检查时一般不易发现。NF-κB的免疫组化和原位杂交测定：与对照组比较,糖耐量受损大鼠造模成功后NF-κB的阳性表达增多,于15周后差异有统计学意义(P<0.05),于20周后有显著差异(P<0.01)。IL-1β的免疫组化和原位杂交测定：与对照组比较,糖耐量受损大鼠造模成功后IL-1β的阳性表达增多,于15周后差异有统计学意义(P<0.05),于20周后有显著差异(P<0.01)。相关性分析：通过Pearson直线相关回归分析发现,实验组大鼠学习记忆成绩与NF-κB、IL-1β阳性表达水平呈负相关,结果有统计学意义(P<0.05)。说明NF-κB、IL-1β表达水平变化与认知障碍有相关性。结论通过水迷宫实验、免疫组化、原位杂交实验等研究,显示糖耐量受损对大鼠认知障碍的发生有显著影响,其发病机制可能与大鼠脑组织中神经细胞炎性因子NF-κB、IL-1β的表达增强有关。更多还原

【Abstract】 ObjectiveIn this study, through the establishment of impaired glucose tolerance rat’s model, detect the cognitive function in rats at different time points, and the expression of key factors in different time points in rat brain tissue inflammatory pathway in rats. We analyzed the relationship between the cognition of rats and the expression of inflammatory factor function. In order to further explore the impaired glucose tolerance effects on cognitive function in rats, and its mechanism. Provide a theoretical basis for our clinical work.MethodWe selected80neonate Wistar rats (body weight180～220g). It will be randomly divided into experimental group and control group, each group40rats. The experimental group was impaired glucose tolerance group. We used high fat diet (add sugar, lard and egg yolk, based on standard full price mixed feed on heat content21.6KJ/kg) feeding the experimental group. We measured blood glucose in rats after5weeks. At5W,10W,15W,20W, water maze was used for assessment of cognition in rats. And we will use to measure nuclear transcription factor in brain tissue of rats with immunohistochemistry and in situ hybridization (factor-kappa B, NF-κB), interleukin-1β(interleukin-1, IL-1β) level. The control group, fed with normal diet, at5W,10W,15W,20W to assess cognitive function and determination of expression of inflammatory factors.ResultsWater maze test: Rats of experimental group at the beginning of the study, there was no significant difference between control group and the ability of learning and memory. With the extension of the feeding time, the rats of the experimental group gradually formed the model, all formed in fifteen weeks of feeding. After repeated monitoring of cognitive function in rats, draw the conclusion: the rats in experimental groups compared with the control group, impaired learning and memory ability, and there are significant differences in fifteenth weeks (P<0.05).HE staining: Under the light microscope, we can occasionally see apoptotic cells were round, red staining cytoplasm, nuclear chromatin aggregation of lumpy. The apoptotic cells are rapidly phagocytosed, and no inflammatory reaction, therefore in routine biopsy is not easy to be found.Determination of immunohistochemistry and in situ hybridization NF-κB: Compared with the control group, we found that the impaired glucose tolerance rats after modeling successfully the positive expression of NF-κB number. After15weeks, the difference was statistically (P<0.05), and in the20weeks was a significant difference (P＜0.01).IL-1βimmunohistochemical assay: Compared with the control group, we found that the tolerance damage in rats after modeling, the positive expression of IL-1βgradually increased.After15weeks, the difference was statistically (P<0.05), and in the20weeks was a significant difference (P<0.01).Correlation analysis:Through Pearson linear correlation and regression analysis we found, the grade of learning and memory was negatively correlated with NF-κB, IL-1βBpositive expression of rats in the experimental group, the results were statistically significant (P<0.05). We found a correlation between expression of NF-κB, IL-1βlevels and cognitive impairment.ConclusionThrough the research on the water maze test, immunohistochemistry, in situ hybridization experiments, the conclusion is glucose tolerance had significant effect on cognitive impairment in rats damaged occurrence. The mechanism may be associated with enhanced expression in rat brain nerve cells, inflammatory factor NF-κB, IL-1β.更多还原