Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Singapore National Medical Research Council Background New treatments are needed to prevent post-angioplasty and stent restenosis in coronary artery disease (CAD) and peripheral arterial disease (PAD) patients. Accumulated findings have shown that mitochondrial dysfunction is involved in the pathophysiology mechanisms underlying several cardiovascular conditions, however, in the scenario of vascular remodelling after injury, the role of mitochondrial dynamics is not completely understood. Purpose In this study, we investigate the effect of mdivi-1 on neointimal hyperplasia and plaque development in a wire-induced endothelial injury animal model.­ Methods Using ApoE-/- mice on a background of hyperlipidaemia, we investigated weather in vivo administration of mdivi-1 (1.2mg/kg/d) could reduce atherosclerotic plaque volume, plaque complexity and inflammation in a carotid-wire injury model of neointimal hyperplasia. Also, THP-1 monocytes (T-M) and THP-1-derived Macrophages (T-DM) were stimulated in vitro with LPS + IFN-gamma to induce M1 polarization and then subjected to gene expression and protein analysis in the presence or absence of mdivi-1 (50µM). Transmigration and 3D chemotaxis assays were performed in T-M cells and Human peripheral blood monocytes, to evaluate the effect of mdivi-1 on cell migration in response to M-CSF and MCP-1. Finally, we investigated the effects of Mdivi-1 on mitochondrial respiration using Seahorse assay in T-M and T-DM. Results In vivo treatment with mdivi-1 reduced neointimal hyperplasia by 37% when compared to control, and was associated with a significant decrease of vascular smooth muscle cells and macrophages cell numbers, as well as reduction of the pro-inflammatory mediators TNF-α and ICAM-1 in the plaque. In vitro M1 polarization of T-M and T-DM induced both up-regulation and production of pro-inflammatory mediators and mdivi-1 significantly suppressed this effects. In addition, monocyte chemotaxis response to M-CSF and MCP-1 was significantly reduced in the presence of mdivi-1. Lastly, mdivi-1 reduced oxygen consumption rate in T-M and T-DM, and prevented M1-cell polarization. Conclusions We show that mdivi-1 significantly reduces vessel wall thickness, neointimal hyperplasia, monocyte recruitment, and inflammation after vascular endothelial injury, positioning Mdivi-1 as a potential pharmacological strategy to reduce restenosis following angioplasty and stenting in CAD and PAD patients.

中文翻译：

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线粒体分裂抑制剂 1 (mdivi-1) 作为预防血管再狭窄的潜在药理学策略

资金致谢 资金来源类型：公共拨款——仅限国家预算。主要资金来源：新加坡国家医学研究委员会 背景需要新的治疗方法来预防冠状动脉疾病 (CAD) 和外周动脉疾病 (PAD) 患者的血管成形术后和支架再狭窄。积累的研究结果表明，线粒体功能障碍参与了几种心血管疾病的病理生理机制，然而，在损伤后血管重塑的情况下，线粒体动力学的作用尚不完全清楚。目的 在本研究中，我们在金属丝诱导的内皮损伤动物模型中研究 mdivi-1 对新生内膜增生和斑块发育的影响。方法在高脂血症背景下使用 ApoE-/- 小鼠，我们研究了 mdivi-1 (1.2mg/kg/d) 的体内天气管理可以减少新内膜增生的颈动脉线损伤模型中的动脉粥样硬化斑块体积、斑块复杂性和炎症。此外，在体外用 LPS + IFN-γ 刺激 THP-1 单核细胞 (TM) 和 THP-1 衍生的巨噬细胞 (T-DM) 以诱导 M1 极化，然后在存在或不存在的情况下进行基因表达和蛋白质分析。 mdivi-1 (50µM)。在 TM 细胞和人外周血单核细胞中进行迁移和 3D 趋化性测定，以评估 mdivi-1 对响应 M-CSF 和 MCP-1 的细胞迁移的影响。最后，我们在 TM 和 T-DM 中使用 Seahorse 测定研究了 Mdivi-1 对线粒体呼吸的影响。结果 与对照相比，mdivi-1 的体内治疗减少了 37% 的新内膜增生，并且与血管平滑肌细胞和巨噬细胞数量的显着减少以及斑块中促炎介质 TNF-α 和 ICAM-1 的减少有关。TM 和 T-DM 的体外 M1 极化诱导促炎介质的上调和产生，而 mdivi-1 显着抑制了这种作用。此外，在存在 mdivi-1 的情况下，单核细胞对 M-CSF 和 MCP-1 的趋化反应显着降低。最后，mdivi-1 降低了 TM 和 T-DM 的耗氧率，并防止了 M1 细胞极化。结论 我们显示 mdivi-1 显着降低血管壁厚度、新内膜增生、单核细胞募集和血管内皮损伤后的炎症，