Department of Cardiology, The Affiliated hospital of Qingdao University, Qingdao, 266000, China.
Institute for translational medicine, Qingdao University, No. 38 Dengzhou Road, 266021, China.
Atherosclerosis. 2020 Apr;298:58-69. doi: 10.1016/j.atherosclerosis.2020.02.018. Epub 2020 Mar 4.
The endothelium is crucially involved in the pathogenesis of atherosclerosis according to accumulating evidence. Moreover, recent studies have showed that lncRNAs could serve as biomarkers of cardiovascular diseases, in particular atherosclerosis. However, the underlying mechanism of endothelial dysfunction involving lncRNAs in atherosclerosis remains unknown. This study investigated the mechanism of lncRNA XXYLT1-AS2 in endothelial dysfunction in atherosclerosis.
The levels of lncRNA XXYLT1-AS2, FUS, VCAM-1, MCP-1, p-AKT, and p-P65 were measured in arteries and HUVEC cell lines via quantitative real-time PCR or Western blot. FISH assay demonstrated that XXYLT1-AS2 and FUS are localized in the nucleus. HUVECs were transfected with si-XXYLT1-AS2 or XXYLT1-AS2 to further assess cell proliferation, migration, and adhesion. Furthermore, bioinformatics analysis, RNA immunoprecipitation and immunofluorescence were performed to investigate the target genes of XXYLT1-AS2 and possible signal pathways.
Overexpression of XXYLT1-AS2 inhibited cell proliferation and migration, reduced the expression of adhesion molecules (VCAM-1) and chemoattractant proteins (MCP-1), and restrained monocyte adhesion to endothelial cells. Mechanistic investigations indicated that XXYLT1-AS2 directly interacts with the target gene FUS/cyclin D1 and modulates the proliferation and migration of endothelial cells (ECs). Moreover, XXYLT1-AS2 exerts a protective role against the inflammatory response in atherosclerosis by blocking NF-κB activity. Clinically, the involvement of XXYLT1-AS2/FUS was also observed in human arteries and the results were consistent with the in vitro analysis.
Our study identified a novel long non-coding RNA (XXYLT1-AS2) and suggests that it might act as an underlying therapeutic target in atherosclerosis-related diseases by regulating ECs functions.
根据不断积累的证据，内皮细胞在动脉粥样硬化的发病机制中起着至关重要的作用。此外，最近的研究表明，lncRNAs 可以作为心血管疾病，特别是动脉粥样硬化的生物标志物。然而，lncRNAs 在内皮功能障碍导致动脉粥样硬化中的潜在机制尚不清楚。本研究探讨了 lncRNA XXYLT1-AS2 在动脉粥样硬化内皮功能障碍中的作用机制。
通过定量实时 PCR 或 Western blot 检测动脉和 HUVEC 细胞系中的 lncRNA XXYLT1-AS2、FUS、VCAM-1、MCP-1、p-AKT 和 p-P65 水平。FISH 实验表明 XXYLT1-AS2 和 FUS 定位于细胞核内。用 si-XXYLT1-AS2 或 XXYLT1-AS2 转染 HUVEC 细胞，进一步评估细胞增殖、迁移和黏附。此外，进行生物信息学分析、RNA 免疫沉淀和免疫荧光实验，以研究 XXYLT1-AS2 的靶基因和可能的信号通路。
XXYLT1-AS2 的过表达抑制细胞增殖和迁移，降低黏附分子（VCAM-1）和趋化因子蛋白（MCP-1）的表达，并抑制单核细胞黏附到内皮细胞。机制研究表明，XXYLT1-AS2 直接与靶基因 FUS/细胞周期蛋白 D1 相互作用，调节内皮细胞（ECs）的增殖和迁移。此外，XXYLT1-AS2 通过阻断 NF-κB 活性，在动脉粥样硬化的炎症反应中发挥保护作用。临床研究中也观察到 XXYLT1-AS2/FUS 在内皮细胞中的作用，结果与体外分析一致。
本研究鉴定了一种新型长非编码 RNA（XXYLT1-AS2），并表明其可能通过调节 ECs 功能，成为与动脉粥样硬化相关疾病的潜在治疗靶点。
