MicroRNAs (miRNAs) have been identified as significant modulators in the pathogenesis of atherosclerosis (AS). Additionally, the dysregulation of vascular smooth muscle cells (VSMCs) is a crucial biological event during AS. Our study aimed to explore the functional roles and molecular mechanisms of miR-141-5p in VSMCs dysfunction. C57BL/6 mice were used to establish AS animal model. Human VSMCs were treated by oxidized low-density lipoprotein (ox-LDL) to establish AS cell model. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to probe miR-141-5p and high-mobility group box 1 (HMGB1) mRNA expressions in VSMCs or plasma samples of the mice. Inflammatory cytokines were detected by enzyme-linked immunosorbent assay kits. Cell counting kit-8 and bromodeoxyuridine assays were performed to evaluate cell proliferation. Cell migration and apoptosis were detected with Transwell assay and flow cytometry analysis, respectively. The target gene of miR-141-5p was predicted with the TargetScan database, and the interaction between miR-141-5p and HMGB1/nuclear factor-κB (NF-κB) was further validated by dual-luciferase reporter assay, qRT-PCR, and Western blot analysis. miR-141-5p was found to be decreased in the plasma of patients and mice model with AS. Its expression was also downregulated in VSMCs treated by ox-LDL. miR-141-5p overexpression inhibited the inflammation, proliferation, migration of VSMCs, and promoted the apoptosis of VSMCs. HMGB1 was identified as a direct target of miR-141-5p, and miR-141-5p could repress the activity of HMGB1/NF-κB signaling. HMGB1 restoration reversed the effects of miR-141-5p, and NF-κB inhibitor JSH-23 showed similar effects with miR-141-5p mimics. miR-141-5p inhibits VSMCs’ dysfunction by targeting the HMGB1/NF-κB pathway, which probably functions as a protective factor during the development of AS.

中文翻译：

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miR-141-5p 通过抑制 HMGB1/NF-κB 通路抑制血管平滑肌细胞炎症、增殖和迁移

MicroRNAs (miRNAs) 已被确定为动脉粥样硬化 (AS) 发病机制中的重要调节剂。此外，血管平滑肌细胞 (VSMC) 的失调是 AS 期间的一个关键生物学事件。我们的研究旨在探讨 miR-141-5p 在 VSMC 功能障碍中的功能作用和分子机制。C57BL/6小鼠用于建立AS动物模型。用氧化低密度脂蛋白（ox-LDL）处理人血管平滑肌细胞，建立AS细胞模型。采用定量实时聚合酶链反应 (qRT-PCR) 来探测小鼠 VSMC 或血浆样品中的 miR-141-5p 和高迁移率组框 1 (HMGB1) mRNA 的表达。通过酶联免疫吸附测定试剂盒检测炎症细胞因子。进行细胞计数试剂盒 8 和溴脱氧尿苷测定以评估细胞增殖。分别用Transwell测定和流式细胞术分析检测细胞迁移和凋亡。用TargetScan数据库预测miR-141-5p的靶基因，并通过双荧光素酶报告基因检测、qRT-PCR进一步验证了miR-141-5p与HMGB1/核因子-κB（NF-κB）的相互作用和蛋白质印迹分析。发现 miR-141-5p 在患有 AS 的患者和小鼠模型的血浆中降低。它的表达也在 ox-LDL 处理的 VSMC 中下调。miR-141-5p过表达抑制了VSMCs的炎症、增殖、迁移，促进了VSMCs的凋亡。HMGB1 被确定为 miR-141-5p 的直接靶标，并且 miR-141-5p 可以抑制 HMGB1/NF-κB 信号传导的活性。HMGB1 恢复逆转了 miR-141-5p 的作用，和 NF-κB 抑制剂 JSH-23 显示出与 miR-141-5p 模拟物相似的效果。miR-141-5p 通过靶向 HMGB1/NF-κB 通路抑制 VSMCs 的功能障碍，该通路可能在 AS 的发展过程中起保护作用。