孕期缺氧对青年子代认知功能及海马NMDAR-Wnt-Catenin通路的影响

发布时间：2018-05-17 01:00

本文选题：孕期 + 缺氧　；参考：《苏州大学》2016年硕士论文

【摘要】：第一部分孕期缺氧对青年子代脑发育及学习记忆等认知功能的影响目的:孕期缺氧可以导致胎儿出生后以至于成年时期出现各种生理病理的改变,对某些成人疾病(Adult diseases)或慢性疾病的易感性增加。本研究探讨孕期缺氧对胚胎期以及青年子代脑发育及功能的长期影响。方法:采用本实验室已建立成熟的孕期缺氧模型,将怀孕母鼠随机分为孕期缺氧组(缺氧组,PH)和常氧组(对照组,Con)。孕期缺氧组母鼠(妊娠第4-21天)放置于10.5%缺氧箱中,对照组母鼠(妊娠第4-21天)放置于21%常氧箱中,出生后子代鼠均在常氧环境下饲养。分别称量胎鼠、6周龄大鼠的体重、脑重。采用Morris水迷宫实验检测6周龄子代大鼠的空间学习记忆能力。结果:孕期缺氧组胎鼠脑重及体重均明显低于常氧对照组;孕期缺氧组6周龄子代大鼠脑重明显低于常氧对照组,而体重并无明显差异。Morris水迷宫实验结果显示,两组6周龄子代大鼠之间游泳速度无明显差异,孕期缺氧子代大鼠在定位航行实验中需要更长的时间才能找到平台;在空间搜索实验中的穿越平台次数明显低于常氧对照组。结论:孕期缺氧不仅直接影响胎儿的生长和脑发育,而且其产生的不良效应可持续到子代青年期,导致青年子代学习记忆等认知能力下降。第二部分孕期缺氧对青年子代海马NMDAR-Wnt-Catenin通路的影响目的:通过分子生物学手段,研究孕期缺氧对子代海马NMDAR-Wnt-Catenin通路的影响。方法:孕期缺氧模型的建立同第一部分;取6周龄子代大鼠的新鲜海马组织,以荧光定量PCR、Western blot方法检测海马组织内NMDAR-Wnt-Catenin通路相关基因(Grin1、Grin2a、Grin2b、Wnt2、Wnt3a、Wnt5a、Wnt7a、β-catenin、Fosl1)及蛋白(NR1、NR2A、NR2B、β-catenin、活化的β-catenin)的表达。结果:荧光定量PCR结果显示,与对照组相比,缺氧组6周龄子代大鼠海马NMDAR相关基因Grin1、Grin2a、Grin2b均下降,其中以Grin2a下降最为明显;Wnt通路相关基因Wnt3a明显下降,但Wnt通路主要受体Fzd4以及LRP5/6无明显变化、β-catenin有下降趋势、β-catenin的下游基因Fosl1明显下降。Western blot结果显示,缺氧组6周龄子代鼠海马组织内的NR1、NR2A、NR2B、活化的β-catenin的蛋白表达量均显著低于对照组,总的β-catenin的蛋白表达无明显变化。结论:孕期缺氧对子代神经系统产生长期不良影响,可通过引起子代海马组织中NMDARs以及Wnt-β-Catenin通路以及下游相关基因的改变导致青年子代的学习记忆等认知功能受损。
[Abstract]:Part one the effect of hypoxia during pregnancy on the cognitive function of brain development, learning and memory of young offspring: hypoxia during pregnancy may lead to various physiological and pathological changes after birth and even in adulthood. Increased susceptibility to certain adult or chronic diseases. The aim of this study was to investigate the long-term effects of hypoxia during pregnancy on brain development and function in embryonic and young offspring. Methods: a mature model of hypoxia during pregnancy was established in our laboratory. Pregnant rats were randomly divided into two groups: hypoxia group (PHH) and normoxic group (control group). The pregnant rats of hypoxia group (4-21 days of pregnancy) were placed in 10.5% hypoxia box, the control group (4-21 days of pregnancy) were placed in 21% normoxic box, and the offspring of postnatal mice were raised in normoxic environment. The body weight and brain weight of 6 weeks old rats were measured respectively. The spatial learning and memory abilities of 6-week-old rats were measured by Morris water maze test. Results: the brain weight and body weight of fetal rats in hypoxia group were significantly lower than those in normoxic control group, and the brain weight in 6-week-old offspring in hypoxia group was significantly lower than that in normoxic control group, but there was no significant difference in body weight. Morris water maze test showed that there was no significant difference between the two groups. There was no significant difference in swimming speed between the two groups of 6-week-old rats. It took a longer time for the hypoxic offspring to find the platform in the navigation experiment, and the frequency of crossing the platform in the spatial search experiment was significantly lower than that in the normoxic control group. Conclusion: hypoxia during pregnancy can not only directly affect the growth and brain development of the fetus, but also produce adverse effects that can be sustained into the youth of the offspring, resulting in the decline of cognitive ability such as learning and memory in the young offspring. Part two effects of hypoxia during pregnancy on NMDAR-Wnt-Catenin pathway in hippocampus of young offspring objective: to study the effect of hypoxia during pregnancy on NMDAR-Wnt-Catenin pathway in hippocampus of young offspring by molecular biology. Methods: the model of anoxia during pregnancy was established in the same part, the fresh hippocampal tissues of 6-week-old rats were collected, and the expression of NMDAR-Wnt-Catenin pathway related genes, Wnt3a1, Wnt5aWnt7a, 尾 -catenininFosl1) and the protein NR1, NR2ANR2B, 尾 -catenin, activated 尾 -catenin, were detected by fluorescence quantitative blot method in hippocampal tissues. The expression of the NMDAR-Wnt-Catenin pathway related genes of Wnt3a1 and Wnt5aWnt7a, 尾 -catenin1) and the protein NR1, NR2ANR2B, 尾 -catenin, activated 尾 -catenin in hippocampal tissues were detected by fluorescence quantitative blot. Results: compared with the control group, the NMDAR related gene Grin1 and Grin2a Grin2b in the hippocampus of the anoxic group were all decreased, and the decrease of Grin2a was the most obvious. However, Fzd4 and LRP5/6, the main receptors of Wnt pathway, did not change, but 尾 -catenin gene Fosl1 decreased significantly. Western blot results showed that the expression of NR1, NR2ANR2B and activated 尾 -catenin protein in hippocampal tissues of 6-week-old filial generation rats in hypoxia group was significantly lower than that in control group. The protein expression of total 尾-catenin did not change significantly. Conclusion: hypoxia during pregnancy has a long-term adverse effect on the nervous system of the offspring, which can result in the impairment of cognitive function such as learning and memory in the young offspring by causing the changes of NMDARs, Wnt- 尾 -Catenin pathway and downstream genes in the hippocampus of the offspring.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R714.2

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