长链非编码RNA(lncRNA)在动脉粥样硬化(As)的发生发展中发挥重要作用，但其机制尚未完全阐明。DNA去甲基化酶TET2在抑制血管内膜增生、抗As过程中发挥重要作用。预实验中我们发现lncRNA TET2-AS1能明显抑制TET2的表达，促进单核-内皮细胞粘附。据此，我们推测“lncRNATET2-AS1通过下调TET2的表达，调控单核细胞与内皮细胞的粘附能力、单核细胞和血管平滑肌细胞的迁移、增殖能力，促进As发生发展”。为证实该假说，项目拟建立lncRNATET2-AS1和TET2过表达的血管细胞模型和沉默lncRNATET2-AS1和TET2表达的apoE敲除鼠As模型，采用基因芯片、实时定量PCR、激光共聚焦和质谱分析等方法，探讨lncRNATET2-AS1参与As形成的分子机制。本研究的完成将从新的视角阐明As的发病机制，为As的有效防治提供新策略和新靶点。

Long chain non-coding RNA(lncRNA)recently have been implicated in many diseases. However, the functional role of lncRNA in atherosclerosis(As) is largely unknown. DNA demethylation modification enzyme ten-eleven translocation2 (TET2) may play important roles in atherosclerosis by inhibition of neointimal hyperplasia in vivo. Preliminary experiments we found that the lncRNA TET2-AS1 can obviously inhibit the expression of TET2 and promote monocyte-endothelial cell adhesion, which imply that lncRNA TET2 AS1-TET2 signaling pathway may play important roles in atherosclerosis. Therefore, we hypothesized that LncRNATET2-AS1,by cutting TET2 expression and regulation of mononuclear cells and endothelial cells adhesion, mononuclear cells and vascular smooth muscle cell migration and proliferation, promote the As development. To test this hypothesis, this study by establishing LncRNATET2-AS1 and TET2 high expression model of the blood vessel cells and silence lncRNATET2-AS1 and TET2 expression of As apoE knockout mice, aims to explore the role of lncRNATET2-AS1-TET2 signaling pathway in As compling with molecular biology techniques, such as gene chip, real time quantitative PCR, laser confocal and mass spectrometry , mass spectrometry.The accomplish of this study will further explore the mechnasim of As with a new perspective, which will provide new interventional strategies and targets for the prevention and treatment for As.

长链非编码RNA(lncRNA)在动脉粥样硬化(As)发生发展中发挥重要作用， TET2可通过表观遗传学调节血管平滑肌分化，影响As发生发展，但其机制尚未完全阐明。本项目研究lncRNA TET2-AS1促细胞粘附的相关机制，同时探讨TET2 在As 发生发展中的作用及机制。研究发现，ox-LDL可促进血管内皮细胞lncRNATET2-AS1表达。lncRNATET2-AS1可抑制血管内皮细胞TET2和eNOS表达，促进ICAM-1、VCAM-1和ET-1表达，从而促进单核细胞与内皮细胞粘附。在高脂饮食喂养的ApoE？/？小鼠，当转染TET2 慢病毒过表达载体时，As斑块面积明显减小，5hmC含量增多，而5mC含量下降。过表达的TET2促进细胞自噬，下调VCAM-1、ICAM-1、MCP-1和IL-1β等炎性因子表达。当转染TET2 shRNA慢病毒载体时，斑块中5hmC含量下降，而5mC含量增多。同时细胞自噬受到抑制，斑块呈现出大脂质核心、巨噬细胞聚积，炎性因子大量表达等不稳定性特征。ox-LDL可抑制内皮细胞自噬。当转染干扰TET2质粒时，ox-LDL预处理的内皮细胞自噬和自噬流进一步受到抑制。当转染TET2过表达质粒时，Beclin 1启动子发生去甲基化，炎性因子表达下调，ox-LDL预处理的内皮细胞自噬和自噬流得到明显改善。总之，我们发现TET2在As形成过程中表达是下调的，下调的TET2促进Beclin 1启动子甲基化，损伤内皮细胞自噬，引起自噬流障碍，上调炎性因子的表达。TET2很可能成为防治As一个新靶点，通过上调TET2表达很可能是防治As一个新策略。