文献类型：期刊文献

中文题名：黑质-纹状体DNA甲基化在6-羟基多巴胺所致帕金森病模型大鼠中的作用

英文题名：Study on DNA methylation of substantia nigra and striatum in the rat model of Parkinson's disease induced by 6-OHDA

作者：梁建庆[1];何建成[2]

第一作者：梁建庆

机构：[1]甘肃中医药大学基础医学院中医诊断教研室,兰州730000;[2]上海中医药大学基础医学院,上海201203

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2018

卷号：43

期号：8

起止页码：636

中文期刊名：解放军医学杂志

外文期刊名：Medical Journal of Chinese People's Liberation Army

收录：CSTPCD;;Scopus;北大核心:【北大核心2017】；CSCD:【CSCD2017_2018】；

基金：国家自然科学基金(81573899;81660767);敦煌医学与转化省部共建教育部重点实验室开放课题(DHYX1516-08);甘肃中医药大学中青年科研基金(ZQ2015-17)~~

语种：中文

中文关键词：帕金森病;DNA甲基化;黑质;纹状体

外文关键词：Parkinson＇s disease;DNA methylation;substantia nigra;corpus striatum

摘要：目的探讨6-羟基多巴胺(6-OHDA)所致帕金森病(PD)模型大鼠黑质-纹状体甲基化状态的变化。方法采用6-OHDA脑立体定向注射术建立大鼠PD模型,另设正常组和假手术组(n=10)。摘取大鼠黑质及纹状体组织,采用ELISA法测定大鼠全基因组甲基化水平;HPLC法检测大鼠黑质及纹状体组织胞嘧啶、5-甲基胞嘧啶含量;Western blotting法检测大鼠黑质及纹状体组织Dnmt1、Dnmt3a、Dnmt3b蛋白表达。采用Methyl RAD技术进行大鼠黑质及纹状体全基因组差异甲基化图谱绘制。结果与正常组及假手术组比较,模型组大鼠黑质及纹状体中DNA总体甲基化水平均明显降低,差异有统计学意义(P<0.01)。与正常组及假手术组比较,模型组大鼠黑质及纹状体中胞嘧啶明显升高、5-甲基胞嘧啶明显降低,差异有统计学意义(P<0.01)。与正常组及假手术组比较,模型组大鼠黑质及纹状体中Dnmt1、Dnmt3a、Dnmt3b蛋白的表达明显增高,差异有统计学意义(P<0.01)。从染色体图谱来看,与正常组及假手术组比较,模型组大鼠黑质及纹状体差异甲基化位点在染色体上的分布不均匀。结论 DNA甲基化修饰异常在帕金森病发病中有一定作用。
Objective To study the changes ofmethylation status ofsubstantia nigra-striatum in the rat model of Parkinson＇s disease （PD） induced by 6-hydroxydopamine （6-OHDA）. Methods PD rats model was established by stereotactic injection of brain with 6-OHDA, the normal control group and the sham operation group were also established （10 rats in each group）. Substantia nigra and striatum of rats were extracted, and ELISA was performed to determine the level of whole genome methylation of rats. The content of cytosine and 5-methylcytosine in substantia nigra and striatum were detected by high performance liquid chromatography and electrochemical detection, and of the expressions ofDnmtl, Dnmt3a and Dnmt3b protein were detected by Western blotting. The differential methylation icon of whole genome of substantia nigra and striatum was plotted using Methyl RAD technique. Results Compared with normal control group and sham operation group, the overall DNA methylation level of substantia nigra and striatum decreased obviously （P〈0.01）, the contents of cytosine increased and of 5-methylcytosine decreased significantly （P〈0.01）, and the expressions of Dnmtl, Dnmt3a and Dnmt3b protein increased markedly with statistical significance （P〈0.01） in model group. The chromosome map indicated that, compared with the normal control group and the sham operation group, an inhomogeneous distribution of different methylation sites of substantia nigra and striatum presented in the model group. Conclusion The abnormality ofDNA methylation modification may play a role in the pathogenesis of Parkinson＇s disease.