circRNA通过调节FoxO1稳定动脉粥样硬化斑块的作用及其分子机制研究

To illustrate whether circRNA_0108265 influences the stability of atherosclerotic plaque and its molecular mechanisms by adjusting FoxO1, the paternal gene, this study is designed to observe the effects of overexpression and silence of circ_0108265, FoxO1 gene and relevant miRNA genes on the secretion of proinflammatory cytokines, collagen metabolism and cell proliferation and apoptosis, respectively. Also,it covers the interaction and molecular mechanisms between circ_0108265 and miRNA, as well as miRNA and FoxO1mRNA. Then, we establish a vulnerable carotid paque model in apolipoprotein E knockout (ApoE-/-) mice by placing a perivascular collar around the carotid artery after high-fat diet feeding for two weeks, and observe the influences of overexpression and silence of circ_0108265, FoxO1 gene on the stability of atherosclerotic plaques.The immplementation of this subject research will illustrate the regulation and molecular mechanisms of circ_0108265 in atherosclerotic plaques and provide new targets for prevention of coronary heart disease.

为了阐明circRNA-circ_0108265是否通过调节父本基因FoxO1影响动脉粥样硬化斑块稳定性及其分子机制，本课题首先在ox-LDL刺激的条件下对细胞进行腺病毒干预，观察circ_0108265过表达或沉默、FoxO1基因过表达或沉默、相关miRNA基因过表达或沉默对细胞炎症因子分泌、胶原代谢、细胞增殖与凋亡等的影响，并研究circ_0108265与miRNA，miRNA与FoxO1mRNA的相互结合及分子机制。然后，在高脂喂养并行颈动脉套管术的ApoE-/-小鼠，观察circ_0108265过表达或沉默、FoxO1基因过表达或沉默对小鼠动脉粥样硬化斑块稳定性的影响。本课题的实施将阐明circ_0108265对动脉粥样硬化斑块的调节作用及分子机制，为冠心病的防治提供新的靶点。

血管内皮细胞异常自噬不利于动脉粥样硬化斑块的稳定。环状RNA (circRNAs)参与调控癌症、代谢和心血管疾病等多种疾病。然而，目前尚不清楚circRNAs是否参与异常内皮细胞自噬和斑块稳定性。我们对70名冠心病患者和30名对照组的外周血单核细胞中的环状RNA进行了序列分析。在冠心病中，共鉴定出2283个下调和85个上调的环状RNA。我们着重研究了冠心病(CHD)相关的hsa_circ_0030042作为内源性真核启动因子4A-III (eIF4A3)海绵，抑制氧化的低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVECs)异常自噬，并保护体内斑块的稳定性。我们发现hsa_circ_0030042通过海绵吸附eIF4A3并阻断其向beclin1和FOXO1 mRNA的聚集来发挥自噬作用，而hsa_circ_0030042引起的beclin1和FOXO1的抑制可以通过eIF4A3的过表达来抵消。在ApoE-/-小鼠中，hsa_circ_0030042过表达也改善了斑块的稳定性，并抵抗了eif4a3诱导的斑块不稳定性。因此，我们提出了一个包括hsa_circ_0030042, eIF4A3, FOXO1和beclin1在内的调控新途径，这可能是一个潜在的治疗冠心病的方法。