Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, People's Republic of China.
Center for Clinical Laboratory, Zhujiang Hospital, Southern Medical University, Guangzhou, 510260, People's Republic of China.
Atherosclerosis. 2023 Aug;379:117183. doi: 10.1016/j.atherosclerosis.2023.06.974. Epub 2023 Jul 8.
The aberrant proliferation and migration of vascular smooth muscle cells (VSMCs) play an essential role in the pathogenesis of atherosclerosis (AS). Long noncoding RNAs (lncRNAs) have been reported as important regulators in a number of diseases. However, very little is known regarding the functional role of lncRNAs in governing proliferation and migration of VSMCs and AS development.
Both in vitro and in vivo assays were performed to investigate the role of lncRNA in the pathophysiology of AS. Our previous lncRNA arrays revealed that lncRNA RP4-639F20.1 was significantly decreased in atherosclerotic plaques. Lentivirus overexpressing RP4-639F20.1 and lncRNA RP4-639F20.1 silencing vectors (Si-lnc-RP4-639F20.1) were constructed and transfected in VSMCs. The in vitro functions of lncRNA were analyzed by CCK-8 assays, EdU assays, scratch wound assays, transwell assays, qRT-PCR and Western blot analyses. RNA fluorescence in situ hybridization, immunoprecipitation and mRNA microarrays were used to explore the underlying mechanism. Adeno-associated-virus-9 (AAV9) overexpressing RP4-639F20.1 was constructed and injected intravenously into ApoE mice to explore the role of lncRNA in vivo.
In vitro experiments showed that lncRNA RP4-639F20.1 interacted with THRAP3 and downregulated c-FOS expression. Both increase of lncRNA RP4-639F20.1 expression and knockdown of c-FOS inhibited the expression of MMP10 and VEGF-α in VSMCs and suppressed VSMCs proliferation and migration. In vivo experiments using ApoE mice fed a high-fat diet demonstrated that lncRNA RP4-639F20.1 overexpression deterred atherosclerosis and decreased lipid levels in atherosclerotic lesions. Patients with coronary artery disease were found to have higher c-FOS levels than healthy individuals and c-FOS expression was positively correlated with the SYNTAX score of patients.
Overall, these data indicated that lncRNA RP4-639F20.1/THRAP3/c-FOS pathway protects against the development of atherosclerosis by suppressing VSMCs proliferation and migration. LncRNA RP4-639F20.1 and c-FOS could represent potential therapeutic targets to ameliorate atherosclerosis-related diseases.
血管平滑肌细胞（VSMCs）的异常增殖和迁移在动脉粥样硬化（AS）的发病机制中起着至关重要的作用。长链非编码 RNA（lncRNA）已被报道为许多疾病的重要调控因子。然而，lncRNA 如何调控 VSMCs 的增殖和迁移以及 AS 的发生发展，目前所知甚少。
通过体外和体内实验研究 lncRNA 在 AS 病理生理学中的作用。我们之前的 lncRNA 芯片显示，lncRNA RP4-639F20.1 在动脉粥样硬化斑块中显著下调。构建了过表达 RP4-639F20.1 的慢病毒和 lncRNA RP4-639F20.1 沉默载体（Si-lnc-RP4-639F20.1），并转染到 VSMCs 中。通过 CCK-8 检测、EdU 检测、划痕愈合实验、Transwell 实验、qRT-PCR 和 Western blot 分析来分析 lncRNA 的体外功能。采用 RNA 荧光原位杂交、免疫沉淀和 mRNA 微阵列来探讨其潜在机制。构建腺相关病毒-9（AAV9）过表达 RP4-639F20.1，并静脉注射到 ApoE 小鼠体内，以研究 lncRNA 在体内的作用。
体外实验表明，lncRNA RP4-639F20.1 与 THRAP3 相互作用，并下调 c-FOS 的表达。lncRNA RP4-639F20.1 的表达增加和 c-FOS 的敲低均抑制了 MMP10 和 VEGF-α 在 VSMCs 中的表达，并抑制了 VSMCs 的增殖和迁移。在高脂饮食喂养的 ApoE 小鼠体内实验中，过表达 lncRNA RP4-639F20.1 可阻止动脉粥样硬化的发生，并降低动脉粥样硬化斑块中的脂质水平。与健康个体相比，冠心病患者的 c-FOS 水平更高，且患者的 c-FOS 表达与 SYNTAX 评分呈正相关。
综上所述，这些数据表明，lncRNA RP4-639F20.1/THRAP3/c-FOS 通路通过抑制 VSMCs 的增殖和迁移来保护动脉粥样硬化的发生。lncRNA RP4-639F20.1 和 c-FOS 可能成为改善动脉粥样硬化相关疾病的潜在治疗靶点。
