Department of Cardiovascular Medicine, The First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China; Department of Cardiovascular Medicine, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, 750004, China.
Department of Cardiovascular Medicine, Shenzhen Hospital of Southern Medical University, Shenzhen, Guangdong, 518100, China.
Eur J Pharmacol. 2022 Jan 15;915:174601. doi: 10.1016/j.ejphar.2021.174601. Epub 2021 Oct 23.
Long non-coding RNAs (LncRNAs) are essential regulators in the occurrence and development of AS. Here we aim to explore the underlying molecular mechanism of LncRNA SNHG16 in regulating ox-LDL-induced VSMC proliferation, migration and invasion. After constructing AS in vivo and in vitro models, the expressions of SNHG16, miR-22-3p, HMBG2, proliferation- and metastasis-related proteins were determined by qRT-PCR and Western blot assays. Detection of serological lipids, H&E and Masson staining analysis were conducted to evaluate the AS injury in mice. The effects of ox-LDL treatment on VSMCs were examined by CCK-8, wound scratch and Transwell Chamber assays. The targeted relationship was measured by luciferase reporter and RIP assays. The results showed that SNHG16 and high-mobility group box 2 (HMGB2) expressions were increased while miRNA-22-3p expression was decreased in AS mice and ox-LDL-stimulated VSMCs. Functionally, sh-SNHG16 restrained ox-LDL-induced VSMC growth and migration. SNHG16 suppressed miRNA-22-3p expression by direct binding. Furthermore, in ox-LDL-treated VSMCs, miRNA-22-3p mimic prevented proliferation, migration, and invasion. Further explorations showed that HMGB2 was a target of miRNA-22-3p, SNHG16 upregulated HMGB2 levels by acting as a competing endogenous RNA (ceRNA) of miRNA-22-3p. More importantly, sh-HMGB2 partially reversed the effects of sh-SNHG16 together with miR-22-2p inhibitor on ox-LDL-induced VSMC proliferation, migration and invasion. Collectively, SNHG16 accelerated atherosclerotic plaque (AP) formation and enhanced ox-LDL-activated VSMCs proliferation and migration by miRNA-22-3p/HMGB2 axis.
长链非编码 RNA（lncRNAs）是 AS 发生和发展的重要调节因子。本研究旨在探讨 lncRNA SNHG16 调节 ox-LDL 诱导的 VSMC 增殖、迁移和侵袭的潜在分子机制。构建 AS 体内和体外模型后，通过 qRT-PCR 和 Western blot 检测 SNHG16、miR-22-3p、HMBG2、增殖和转移相关蛋白的表达。通过酶联免疫吸附测定（ELISA）和 H&E、Masson 染色分析评估小鼠 AS 损伤。通过 CCK-8、划痕和 Transwell 实验检测 ox-LDL 处理对 VSMCs 的影响。通过荧光素酶报告和 RIP 实验检测靶向关系。结果表明，在 AS 小鼠和 ox-LDL 刺激的 VSMCs 中，SNHG16 和高迁移率族蛋白 2（HMGB2）表达增加，而 miR-22-3p 表达降低。功能上，sh-SNHG16 抑制 ox-LDL 诱导的 VSMC 生长和迁移。SNHG16 通过直接结合抑制 miR-22-3p 的表达。此外，在 ox-LDL 处理的 VSMCs 中，miR-22-3p 模拟物可防止增殖、迁移和侵袭。进一步探索表明，HMGB2 是 miR-22-3p 的靶标，SNHG16 通过作为 miR-22-3p 的竞争性内源 RNA（ceRNA）而上调 HMGB2 水平。更重要的是，sh-HMGB2 部分逆转了 sh-SNHG16 与 miR-22-2p 抑制剂联合对 ox-LDL 诱导的 VSMC 增殖、迁移和侵袭的作用。总之，SNHG16 通过 miRNA-22-3p/HMGB2 轴加速动脉粥样硬化斑块（AP）形成，并增强 ox-LDL 激活的 VSMCs 增殖和迁移。
