程序性坏死在铝致斑马鱼阿尔茨海默病模型中的作用研究

发布时间：2018-04-13 12:17

本文选题：氯化铝 + 斑马鱼　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:通过染铝建立斑马鱼阿尔茨海默病模型,探讨程序性坏死在铝致斑马鱼阿尔茨海默病模型中的作用。方法:选取8月龄TU系成年斑马鱼,随机分为4组,每组25条。在养殖液中加入不同浓度的结晶氯化铝(PH6.4)染毒30天:对照组(0μg/L)、低浓度组(20μg/L)、中浓度组(100μg/L)、高浓度组(500μg/L)。染毒结束后,通过斑马鱼T迷宫检测各组斑马鱼学习记忆能力;采用石墨炉原子吸收光谱法检测各组斑马鱼脑铝含量;采用酶联免疫吸附法(ELISA)检测各组斑马鱼脑组织中APP、Aβ1-42、Aβ1-40蛋白含量;采用RT-PCR法检测各组斑马鱼脑组织中m TOR信号通路、阿尔茨海默病和程序性坏死相关基因的表达。结果:1.斑马鱼T迷宫结果:中浓度组和高浓度组斑马鱼在实验第2天、第3天、第4天的潜伏期与对照组相比明显延长(P0.05);高浓度组斑马鱼在实验的第2天、第3天、第4天在EC区停留的总时间与对照组相比减少(P0.05);中浓度组斑马鱼在实验的第3天、第4天在EC区停留的总时间与对照组相比减少(P0.05)。2.斑马鱼脑铝含量检测结果:中浓度组和高浓度组斑马鱼脑铝含量与对照组和低浓度组相比显著升高(P0.05);高浓度组斑马鱼脑铝含量与中浓度组相比显著升高(P0.05)。3.酶联免疫吸附法(ELISA)检测结果:中浓度组和高浓度组斑马鱼脑组织中APP蛋白含量与对照组相比显著增加(P0.05);高浓度组斑马鱼脑组织中Aβ1-42蛋白含量与对照组相比显著增加(P0.05);各组斑马鱼脑组织中Aβ1-40蛋白含量差别无统计学意义(P0.05)。4.RT-PCR检测结果:中浓度组、高浓度组斑马鱼脑组织中m TOR、AKT2的基因表达与对照组、低浓度组相比显著下降(P0.05);高浓度组斑马鱼脑组织中AKT1、PI3K的基因表达与对照组相比均显著下降(P0.05)。中浓度组、高浓度组斑马鱼脑组织中appb、mapta的基因表达与对照组相比显著增加(P0.05);高浓度组斑马鱼脑组织中appb的基因表达与中浓度组相比显著增加(P0.05);中浓度组、高浓度组斑马鱼脑组织中BDNF的基因表达与对照组、低浓度组相比显著下降(P0.05);中浓度组斑马鱼脑组织中BDNF的基因表达与对照组相比显著下降(P0.05)。高浓度组斑马鱼脑组织中RIP1、PARP2、Bmf1、jph3、rab25和caspase8的基因表达与对照组相比均显著增加(P0.05);中浓度组斑马鱼脑组织中PARP2、Bmf1和rab25的基因表达与对照组相比均显著增加(P0.05);各组斑马鱼脑组织中FADD和mag的基因表达差异无统计学意义(P0.05)。结论:1.铝致斑马鱼阿尔茨海默病模型建立成功;2.铝可引起RIP1、PARP2、Bmf1、jph3、rab25和caspase8的基因表达增加;3.程序性坏死在铝致斑马鱼阿尔茨海默病模型中发挥着重要的作用。
[Abstract]:Aim: to establish Alzheimer's disease model of zebrafish by aluminum exposure and to explore the role of programmed necrosis in aluminum-induced Alzheimer's disease model of zebrafish.Methods: 8 months old TU adult zebrafish were randomly divided into 4 groups, 25 in each group.After exposure to zebrafish, the learning and memory ability of zebrafish in each group was measured by T-maze, and the brain aluminum content of zebrafish in each group was detected by graphite furnace atomic absorption spectrometry (GFAAS).Enzyme linked immunosorbent assay (Elisa) was used to detect the protein content of APP A 尾 1-42 A 尾 1-40 in the brain tissues of zebrafish, and the expression of m TOR signaling pathway, Alzheimer's disease and programmed necrosis related genes in the brain tissues of zebrafish were detected by RT-PCR method.The result is 1: 1.Zebrafish T-maze results: the incubation period of zebrafish in the middle and high concentration groups was significantly longer than that in the control group on the 2nd, 3rd and 4th day, while in the high concentration group, the latency was significantly prolonged on the 2nd and 3rd day of the experiment.On the 4th day, the total stay time of zebrafish in EC region was lower than that in control group, and the total stay time of zebrafish in the middle concentration group was lower than that in the control group on the 3rd day and the 4th day, compared with the control group.The results showed that the brain aluminum content of zebrafish in the middle and high concentration groups was significantly higher than that in the control group and the low concentration group, and that in the high concentration group was significantly higher than that in the middle concentration group and that in the middle concentration group, and that in the high concentration group was significantly higher than that in the middle concentration group.Enzyme linked immunosorbent assay (Elisa): the content of APP protein in the brain tissue of zebrafish in middle and high concentration groups was significantly higher than that in control group, and the content of A 尾 1-42 protein in brain tissue of zebrafish in high concentration group was significantly higher than that in control group.There was no significant difference in A 尾 1-40 protein content in the brain tissue of zebrafish in each group. 4. Results of RT-PCR: middle concentration group;Compared with the control group, the expression of mTORA AKT2 gene in the brain tissue of the high concentration group was significantly lower than that of the control group, and that of the high concentration group was significantly lower than that of the control group, while the gene expression of PI3K in the brain tissue of the high concentration group was significantly lower than that of the control group.In the middle and high concentration groups, the expression of appb gene in the brain tissue of zebrafish was significantly higher than that of the control group, while the expression of appb gene in the brain tissue of the high concentration group was significantly higher than that of the middle concentration group, while that of the middle concentration group was significantly higher than that of the middle concentration group, while that of the middle concentration group was significantly higher than that of the control group.The expression of BDNF gene in the brain tissue of zebrafish in high concentration group was significantly lower than that in control group, and the expression of BDNF gene in brain tissue of zebrafish in middle concentration group was significantly lower than that in control group.There was no significant difference in gene expression between FADD and mag.Conclusion 1.The model of Alzheimer's disease in zebrafish induced by aluminum was established successfully.Aluminum can increase the gene expression of RIP1, PARP2, Bmf1, jph3, rab25 and caspase8.Programmed necrosis plays an important role in aluminum-induced Alzheimer's disease model of zebrafish.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.16;R-332

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