Qiao Shan, Wang Xing, Li Haiyun, Zhang Canling, Wang Aihua, Zhang Shanchao
Department of Neurology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong First Medical University, Jinan, China; Department of Neurology, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan, China; Department of Medical Genetics, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China; Shandong Laibo Biotechnology Co., Ltd, China.
Department of Neurology, Tianyou Affiliated Hospital, Wuhan University of Science and Technology, Wuhan, China.
Nutr Metab Cardiovasc Dis. 2023 Aug;33(8):1619-1631. doi: 10.1016/j.numecd.2023.05.007. Epub 2023 May 11.
Atherosclerosis (AS) is a chronic inflammatory disease that damages the arterial wall as a result of hyperlipidemia and causes endothelial cell dysfunction, which increases the risk of atherothrombotic events. Multiple pathological conditions have shown ectopic miR-199a-5p levels to cause endothelial injury, but its role in the AS competitive endogenous RNA (CeRNA) network is still unknown.
The high-fat diet (HFD) apoE-/- mouse model was constructed in vivo, and ECs were cultured under ox-LDL treatment to induce EC injury in vitro. Immunohistochemistry and immunofluorescence staining were used to assess the effect of miR-199a-5p on the macrophage, SMC, collagen content, and endothelial coverage in the artery wall of mouse model. miR-199a-5p level was validated to be overexpression in the aorta tissue of HFD apoE-/- mice and in the ox-LDL-treated ECs, and even in the plasma EVs of the patients with cerebral AS. Silencing of miR-199a-5p significantly attenuated atherosclerotic progress in HFD apoE-/- mice, and the gain/loss-of-function assay indicated that miR-199a-5p overexpression aggravated ox-LDL-induced disabilities of endothelial proliferation, motility, and neovascularization based on cell counting kit-8 assay, transwell assay and matrigel assay. Mechanistically, miR-199a-5p prevented EC activation by activating the FOXO signaling pathway by targeting SIRT1. Additionally, circular RNA (circRNA) circHIF1ɑ was identified as having a low expression in the ox-LDL-treated EC and mediated SIRT1 expression via sponging miR-199a-5p to rescue ox-LDL-induced EC injury.
Our study demonstrated the vital role of miR-199a-5p/SIRT1 axis regulated by circHIF1ɑ in AS pathogenesis and provided novel effective targets for AS treatment.
动脉粥样硬化（AS）是一种慢性炎症性疾病，由于高脂血症会损害动脉壁并导致内皮细胞功能障碍，进而增加动脉粥样硬化血栓形成事件的风险。多种病理状况已表明异位的miR-199a-5p水平会导致内皮损伤，但其在AS竞争性内源性RNA（CeRNA）网络中的作用仍不清楚。
在体内构建高脂饮食（HFD）载脂蛋白E基因敲除（apoE-/-）小鼠模型，并在体外对内皮细胞（ECs）进行氧化型低密度脂蛋白（ox-LDL）处理以诱导EC损伤。采用免疫组织化学和免疫荧光染色评估miR-199a-5p对小鼠模型动脉壁中巨噬细胞、平滑肌细胞（SMC）、胶原含量和内皮覆盖的影响。经证实，miR-199a-5p在HFD apoE-/-小鼠的主动脉组织、ox-LDL处理的ECs中，甚至在脑AS患者的血浆细胞外囊泡（EVs）中均呈过表达。沉默miR-199a-5p可显著减轻HFD apoE-/-小鼠的动脉粥样硬化进展，功能获得/丧失实验表明，基于细胞计数试剂盒-8实验、Transwell实验和基质胶实验，miR-199a-5p过表达会加重ox-LDL诱导的内皮细胞增殖、迁移和新生血管形成障碍。机制上，miR-199a-5p通过靶向沉默调节蛋白1（SIRT1）激活叉头框蛋白O（FOXO）信号通路来阻止EC激活。此外，环状RNA（circRNA）circHIF1ɑ在ox-LDL处理的EC中被鉴定为低表达，并通过海绵吸附miR-199a-5p介导SIRT1表达以挽救ox-LDL诱导的EC损伤。
我们的研究证明了由circHIF1ɑ调控的miR-199a-5p/SIRT1轴在AS发病机制中的重要作用，并为AS治疗提供了新的有效靶点。
