糖耐量受损大鼠认知障碍与脑组织中炎症因子表达的相关性研究

发布时间：2018-03-17 10:19

本文选题：糖耐量受损　切入点：炎症反应　出处：《天津医科大学》2014年硕士论文　论文类型：学位论文

【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(P0.05)。 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(P0.05),于20周后有显著差异(P0.01)。 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(P0.05),于20周后有显著差异(P0.01)。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(P0.05)。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。
[Abstract]:Purpose. In this study, the rat model of impaired glucose tolerance was established to detect the cognitive function of rats at different time points and the expression of key factors in inflammatory pathway of rat brain at different time points. The relationship between cognitive function and the expression of inflammatory factors in rats was analyzed in order to further explore the effects of impaired glucose tolerance on cognitive function and its related mechanisms in order to provide theoretical basis for guiding clinical work. Method. Eighty newborn Wistar rats (body weight 180-220 g) were randomly divided into experimental group and control group with 40 rats in each group. The experimental group was treated with high fat and high sugar diet. Experimental animals and feed were provided by the Animal experiment Center of Tianjin Medical University for blood glucose determination after 5 weeks. The cognition of rats was assessed by water maze method at 5 weeks, 10 weeks, 15 weeks, and 20 weeks, respectively. The levels of factor-kappa B, NF- 魏 B, interleukin-1 尾 -interleukin-1 尾, IL-1 尾 in rat brain were determined by immunohistochemistry and in situ hybridization. The cognitive function was assessed and the expression of inflammatory factors was measured. Results. Water maze test: there was no significant difference in learning and memory ability between the experimental group and the control group at the beginning of the experiment. With the prolongation of the feeding time, the rats of the experimental group gradually became models, and all of the rats in the experimental group became models at the 15th week of feeding. After repeated monitoring of the cognitive function of the experimental group compared with the control group, the learning and memory abilities of the experimental group were decreased, and there was a significant difference after 15 weeks (P 0.05). HE staining: compared with the control group, apoptosis cells were occasionally seen in the experimental group under light microscope, cytoplasmic red staining, nuclear chromatin aggregation into a mass, because the apoptotic cells were quickly swallowed up, and there was no inflammatory reaction. Therefore, it is not easy to find in the routine section examination. Immunohistochemistry and in situ hybridization of NF- 魏 B: compared with the control group, the positive expression of NF- 魏 B was increased in the model of impaired glucose tolerance rats, which was significantly different after 15 weeks and 20 weeks later. Immunohistochemical and in situ hybridization of IL-1 尾: compared with the control group, the positive expression of IL-1 尾 increased after successful modeling in impaired glucose tolerance rats. The difference was statistically significant after 15 weeks, and significantly different after 20 weeks. Correlation analysis: by Pearson linear correlation regression analysis, it was found that there was a negative correlation between the learning and memory achievement and the positive expression of NF- 魏 B, IL-1 尾 in the experimental group, the results showed that there was a significant correlation between the expression of IL-1 尾 and cognitive impairment in the experimental group (P 0.05). Conclusion. The results of water maze test, immunohistochemistry and in situ hybridization showed that impaired glucose tolerance had a significant effect on the occurrence of cognitive impairment in rats. The pathogenesis may be related to the increased expression of neuroinflammatory factor NF- 魏 B and IL-1 尾 in rat brain.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R749.24

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