Genonic analysis to Identify atherosclerosis-susceptible and -resistant genes and Investigation of molecular mechanism In the pathogenesis of atherosclerosis

本项目的具体目标是(1)通过分析动脉粥样硬化易感和抵抗小鼠来识别动脉粥样硬化的易感和抵抗基因，以及(2)利用候选基因和新发现的动脉粥样硬化易感和抵抗基因的单核苷酸多态性(SNPs)进行遗传分析。1.动脉粥样硬化易感和抵抗基因的鉴定为了鉴定动脉粥样硬化易感和抵抗基因，用高胆固醇饲料喂养动脉粥样硬化易感小鼠(C57BL/6J)和动脉粥样硬化抵抗小鼠(C3H/HeJ)。随后，从这些动物的血管内皮细胞、血管平滑肌细胞和骨髓干细胞中提取mRNAs用于表达谱(在viva表达谱中)。进行了表情分析，目前正在分析收集到的数据。同时，对生成PRI…进行了优化2.动脉粥样硬化相关候选基因序列变异的鉴定在几个候选基因中确定了MCP-1 A-2518G和CCR2 G270A多态性的实验条件。然后搜索候选基因MCP-1、转化生长因子-β和核因子-κB的多态，但没有发现改变基因功能的新的多态。与此同时，对缺血性心脏病(IHD)患者的招募仍在继续，但300名IHD患者中只有2人被发现。重新分析我们的患者数据库，发现冠状动脉危险因素的数量对动脉粥样硬化的发病机制影响很小。因此，改变了患者招募的策略，并开始以新的标准招募IHD患者。未来，将利用动脉硬化相关候选基因的多态性进行遗传分析。较少

Specific aims of this project are (1)to Identify atherosclerosis-susceptible and -resistant genes by analyzing atherosclerosis-susceptible and -resistant mice, and (2)to carry out genetic analysis by using single nucleotide polymorphisms (SNPs) of candidate genes and newly identified atherosclerosis-susceptible and -resistant genes. Progress of this project is as follows;1. Identification of atherosclerosis-susceptible and -resistant genesTo identity atherosclerosis-susceptible and -resistant genes, atherosclerosis-susceptible mouse line (C57BL/6J) and atherosclerosis -resistant mouse line (C3H/HeJ), were fed with high cholesterol diet. Subsequently, mRNAs were extracted from vascular endothelial cells, vascular smooth muscle cells and bone marrow stem cells of these animals for expression profiling (In viva expression profiling) with cDNA microarray chips. Expression profiling was carried out, and collected data are now under analysis. At the same time, optimization for generating pri … More mary cultured cells from vascular endothelial cells, vascular smooth muscle cells and bone marrow stem cells of these animals are undergone to perform in vitro expression profiling.2.Identification of sequence variations in atherosclerosis-related candidate genes for genetic analysisAmong several candidate genes experimental condition to determine MCP-1 A-2518G polymorphism and CCR2 G270A polymorphism has been set. Then, polymorphisms of candidate genes, such as MCP-1,TGF-β and NF-κB, were searchd, but no novel polymorphisms to alter gene function were identified. At the same time, recruitment of ischemic heart diseasee(IHD)patients has been continued,but only 2 out of 300 IHD patients were found. Our patient database was reanalyzed, and It was identified that number of coronary risk factors had an small influence on the pathogenesis of atherosclerosis. Therefore, a strategy of patient recruitment was changed, and recruitment of IHD patients with new criteria was begun. ln future, genetic analysis using polymorphisms of aterosclerosis-related candidate genes will be conducted. Less