BACKGROUND Atherosclerosis is the main cause of cardiovascular diseases such as ischemic stroke and coronary heart disease. Gene-specific promoter methylation changes have been suggested as one of the causes underlying the development of atherosclerosis. We aimed to identify and validate specific genes that are differentially expressed through promoter methylation in atherosclerotic plaques. We performed the present study in four steps: (1) profiling and identification of gene-specific promoter methylation changes in atherosclerotic tissues; (2) validation of the promoter methylation changes of genes in plaques by comparison with non-plaque intima; (3) evaluation of promoter methylation status of the genes in vascular cellular components composing atherosclerotic plaques; and (4) evaluation of promoter methylation differences in genes among monocytes, T cells, and B cells isolated from the blood of ischemic stroke patients. RESULTS Upon profiling, AIRE1, ALOX12, FANK1, NETO1, and SERHL2 were found to have displayed changes in promoter methylation. Of these, AIRE1 and ALOX12 displayed higher methylation levels in plaques than in non-plaque intima, but lower than those in the buffy coat of blood. Between inflammatory cells, the three genes were significantly less methylated in monocytes than in T and B cells. In the vascular cells, AIRE1 methylation was lower in endothelial and smooth muscle cells. ALOX12 methylation was higher in endothelial, but lower in smooth muscle cells. Immunofluorescence staining showed that co-localization of ALOX12 and AIRE1 was more frequent in CD14(+)-monocytes than in CD4(+)-T cell in plaque than in non-plaque intima. CONCLUSIONS Promoter methylation changes in AIRE1 and ALOX12 occur in atherosclerosis and can be considered as novel epigenetic markers.

中文翻译：

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ALOX12 和 AIRE1 中的启动子甲基化变化：动脉粥样硬化的新型表观遗传标记。

背景动脉粥样硬化是心血管疾病如缺血性中风和冠心病的主要原因。基因特异性启动子甲基化变化被认为是动脉粥样硬化发展的潜在原因之一。我们旨在鉴定和验证通过动脉粥样硬化斑块中启动子甲基化差异表达的特定基因。我们分四个步骤进行了本研究：（1）分析和鉴定动脉粥样硬化组织中基因特异性启动子甲基化变化；(2)与非斑块内膜相比，验证斑块中基因的启动子甲基化变化；(3) 评估构成动脉粥样硬化斑块的血管细胞成分中基因的启动子甲基化状态；(4) 评估从缺血性中风患者血液中分离的单核细胞、T 细胞和 B 细胞之间基因的启动子甲基化差异。结果 分析后，发现 AIRE1、ALOX12、FANK1、NETO1 和 SERHL2 显示启动子甲基化的变化。其中，AIRE1 和 ALOX12 在斑块中的甲基化水平高于非斑块内膜，但低于血沉棕黄层中的甲基化水平。在炎症细胞中，这三种基因在单核细胞中的甲基化程度明显低于 T 细胞和 B 细胞。在血管细胞中，内皮细胞和平滑肌细胞中的 AIRE1 甲基化较低。ALOX12 甲基化在内皮细胞中较高，但在平滑肌细胞中较低。免疫荧光染色显示，与非斑块内膜相比，斑块中 ALOX12 和 AIRE1 的共定位在 CD14(+)-单核细胞中比在 CD4(+)-T 细胞中更频繁。结论 AIRE1 和 ALOX12 的启动子甲基化变化发生在动脉粥样硬化中，可被视为新的表观遗传标记。