线粒体外膜电压依赖阴离子通道VDAC差异表达在母系遗传性高血压发病中的调控机制研究

项目名称： 线粒体外膜电压依赖阴离子通道VDAC差异表达在母系遗传性高血压发病中的调控机制研究

项目编号： No.81470542

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 刘昱圻

作者单位： 中国人民解放军总医院

项目金额： 73万元

中文摘要： 原发性高血压（EHT）是导致脑卒中、冠心病和晚期肾功能衰竭等疾病的独立危险因素。EHT受环境因素和遗传因素等多因素影响，近来研究表明母系遗传性高血压与线粒体基因（mtDNA）突变有关，但其具体作用机制尚未有系统研究报道。本课题通过高血压风险评估模型和线粒体功能验证筛查和发现mtDNA突变相关的高血压患者，分离血管平滑肌细胞和外周血内皮祖细胞，分别构建血管平滑肌细胞系和诱导分化内皮细胞系，以线粒体外膜电压依赖阴离子通道（Voltage-dependent anion channel, VDAC）为研究靶点，利用RT-PCR测序和双向凝胶电泳对比研究不同细胞系VDAC基因和蛋白表达差异，并利用VDAC选择性阻断剂和ROS干预下，观察对线粒体基质内钙、细胞内钙以及线粒体膜电势的影响，并利用荧光共振能量转移（FRET）技术观察VDAC与线粒体内膜钙离子通道MCU之间动态相互作用，初步探讨mtDNA突变在高血压发病中的作用机制，为具有母系遗传的原发性高血压早期诊断提供理论依据。

中文关键词： 原发性高血压；线粒体；电压依赖阴离子通道；钙离子通道；基因突变

英文摘要： Essential hypertension （EHT）is an independent risk factor for stroke, coronary heart disease and end-stage renal failure and other diseases. EHT is influenced by multiple factors such as environmental factors and genetic factors. Recently, some studies have shown that maternally inherited hypertension was associated with mitochondrial DNA (mtDNA) mutations, but the mechanism is still unclear. In this study, EHT patients associated with mtDNA were identified and verified by mitochondrial function evaluation. The vascular smooth muscle cells and endothelial progenitor cells (EPC) were isolated and induced to endothelial cells. Differential expression of voltage-dependent anion channel (VDAC) located at mitochondrial outer membrane were studied with RT-PCR and two-dimensional gel electrophoresis. To observe the change of [Ca2+]m, [Ca2+]c and mitochondrial membrane potential with influenced by VDAC selective blockers and ROS intervention. The fluorescence resonance energy transfer (FRET) technique was used to observe the dynamic interaction between VDAC and endometrial calcium channel MCU. The pathogenesis mechanism of mtDNA mutations in maternally inherited hypertension is initially discussed, which provide a theory for early diagnosis and genes intervention of maternally inherited hypertension.

英文关键词： hypertension;mitochondrial;VDAC;calcium channel;gene mutation