SIRT1介导组蛋白乙酰化在同型半胱氨酸致动脉粥样硬化中的作用及特异性miRNAs调控机制

项目名称： SIRT1介导组蛋白乙酰化在同型半胱氨酸致动脉粥样硬化中的作用及特异性miRNAs调控机制

项目编号： No.81460080

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 杨晓明

作者单位： 宁夏医科大学

项目金额： 47万元

中文摘要： 动脉粥样硬化(As)是以脂代谢紊乱为特征的全身性疾病，前期研究同型半胱氨酸(Hcy)致As时发现不同基因DNA高低甲基化并存，提示存在更深层次机制；组蛋白乙酰化具有协同DNA甲基化调控基因转录的作用，但Hcy是否通过组蛋白乙酰化异常致As未见报道，而miRNAs是基因表达调控的重要方式，因此本课题拟复制HHcy As模型，免疫印迹检测组蛋白乙酰化的改变，阐明H3和H4乙酰化在As中的作用；定量PCR等检测HDACs、HATs及SIRT1的表达，构建针对SIRT1的过表达和抑制表达载体转染，阐明组蛋白乙酰化的机制并确定SIRT1是关键靶基因；运用微阵列技术筛选Hcy特异性miRNAs，转染miRNAs抑制物和过表达载体，观察其对组蛋白乙酰化的影响；构建携载SIRT1重组质粒及突变荧光素酶报告质粒并转染，探讨特异性miRNAs靶向调控SIRT1的机制，确定关键靶点，为As靶向治疗提供理论依据。

中文关键词： 同型半胱氨酸；动脉粥样硬化；组蛋白乙酰化；沉默信息调节因子1

英文摘要： Atherosclerosis (As) is a systemic disease which is characterized by lipid metabolism disorder. Previous research has shown that hypermethylation and hypomethylation were co-exist in the process of Hcy induced As,which indicates the existence of deeper regulatory mechanism. Histone acetylation regulates genetic transcription coordinate with DNA methylation, however, it has not been reported that whether Hcy causes As through abnormal histone acetylation and miRNAs is an important way in regulating gene expression；Therefore, this project intends to replicate the model of HHcy-induced As, detecting the changes of histone acetylation with western blot to clarify the role of H3 and H4 acetylation in As; detecting the expression of HDACs, HATs, and SIRT1 by qRT-PCR. Constructing the over-expressed and suppressive vectors of SIRT1 to clarify the mechanism of histone acetylation and determine whether SIRT1 is the key target gene. Microarray is used to screen Hcy specific miRNAs, and observe its effect on histone acetylation after transfecting the over-expressed and suppressive vectors of the specific miRNAs. Constructing the recombinant plasmid carrying SIRT1 and mutant luciferase reporter plasmid and transfect into related cells to discuss the mechanism of specific miRNAs target regulating SIRT1 and identify the key targets, to provide a theoretical basis for the targeted therapy of As.

英文关键词： Homocysteine;Atherosclerosis;Histone acetylation;Silent mating type information regulation 2 homolog 1