MPTP诱导的帕金森病小鼠模型黑质脑组织DNA甲基化研究

发布时间：2018-01-20 00:10

本文关键词： 帕金森病 DNA甲基化总体甲基化 DNA甲基转移酶泛素羧基末端水解酶　出处：《生物化学与生物物理进展》2015年03期 　论文类型：期刊论文

【摘要】：为研究DNA甲基化在帕金森病发病机制中的作用,本研究用环境毒素1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)连续腹腔给药诱导小鼠帕金森病(Parkison's disease,PD)模型,应用ELISA检测小鼠黑质脑组织总体甲基化水平,应用实时荧光定量PCR方法检测DNA甲基转移酶表达水平,探讨MPTP诱导的小鼠PD模型黑质部位是否存在DNA甲基化异常.进一步应用甲基化DNA免疫共沉淀结合DNA甲基化芯片方法,构建MPTP诱导的小鼠PD模型黑质脑组织DNA甲基化谱,并寻找DNA甲基化修饰异常的PD相关基因对其进行验证.结果表明,模型组小鼠黑质脑组织DNA总体甲基化水平较对照组显著降低,Dnmt1的表达水平显著增高.利用DNA甲基化芯片在全基因组内筛选出甲基化差异修饰位点共48个,涉及44个基因,这些甲基化差异基因参与信号转导、分子转运、转录调控、发育、细胞分化、凋亡调控、氧化应激、蛋白质降解等生物学过程.在甲基化差异修饰基因中,对Uchl1基因及Arih2基因进行了甲基化水平以及表达水平的验证.结果表明,模型组小鼠黑质脑组织Uchl1启动子区域甲基化水平较对照组增高,m RNA及蛋白质表达水平降低,Arih2启动子区域甲基化水平较对照组降低,m RNA及蛋白质表达水平增高.实验结果进一步证实,DNA甲基化修饰异常在帕金森病发病机制中有重要作用,环境因素(如MPTP)可以通过改变DNA甲基化修饰参与帕金森病的发生发展.
[Abstract]:In order to study the role of DNA methylation in the pathogenesis of Parkinson's disease, the environmental toxin 1-methyl-4-phenyl-1tian3 was used in this study. 6-Tetrahydropyridine (MPTP) was used to induce the model of Parkinson's disease (PD) in mice. The total methylation level in the substantia nigra of mice was detected by ELISA and the expression of DNA methyltransferase was detected by real-time fluorescence quantitative PCR. To investigate the presence of abnormal DNA methylation in the substantia nigra of MPTP induced PD mice, the methylated DNA immunoprecipitation combined with DNA methylation chip method was used to detect the presence of DNA methylation in the substantia nigra. To construct DNA methylation spectrum of substantia nigra in MPTP induced PD model and to identify PD-related genes with abnormal DNA methylation modification. The total methylation level of DNA in the substantia nigra of the model group was significantly lower than that in the control group. The expression level of Dnmt1 was significantly increased. A total of 48 methylation differentially modified sites were screened in the whole genome using DNA methylation microarray, involving 44 genes. These methylation differential genes are involved in signal transduction, molecular transport, transcriptional regulation, development, cell differentiation, apoptosis regulation, oxidative stress, protein degradation and other biological processes. The methylation level and expression level of Uchl1 gene and Arih2 gene were verified. The methylation level of Uchl1 promoter region in substantia nigra of the model group was higher than that of the control group. The level of protein expression and the methylation level of the Uchl1 promoter region in the model group were lower than those in the control group. The expression level of m RNA and protein was increased. The results further confirmed that abnormal methylation of RNA may play an important role in the pathogenesis of Parkinson's disease. Environmental factors such as MPTP can be involved in the development of Parkinson's disease by modifying DNA methylation.
【作者单位】：中南大学湘雅医院老年病学神经内科;中南大学湘雅医院神经内科;神经退行性疾病湖南省重点实验室;中南大学医学遗传学国家重点实验室;
【基金】：国家自然科学基金国际(地区)合作与交流(81361120404);国家自然科学基金青年基金(81401060)资助项目国家重点基础研究发展计划(973)(2011CB510000)~~
【分类号】：R742.5;R-332
【正文快照】： 帕金森病(Parkison's disease,PD)是一种常见的神经退行性疾病,在中国65岁以上的老年人中患病率约为1%~2%,且随着年龄增长有增加的趋势[1].PD的发病机制十分复杂,目前认为与遗传因素和环境因素的相互作用有关.最新研究表明,表观遗传学机制在神经退行性疾病中有重要作用.表观遗

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