Welten Sabine M J, de Jong Rob C M, Wezel Anouk, de Vries Margreet R, Boonstra Martin C, Parma Laura, Jukema J Wouter, van der Sluis Tetje C, Arens Ramon, Bot Ilze, Agrawal Sudhir, Quax Paul H A, Nossent A Yaël
Department of Surgery, Leiden University Medical Center, Leiden, The Netherlands; Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, Leiden, The Netherlands.
Department of Surgery, Leiden University Medical Center, Leiden, The Netherlands; Division of Biopharmaceutics, LACDR, Leiden University, Leiden, The Netherlands.
Atherosclerosis. 2017 Jun;261:26-36. doi: 10.1016/j.atherosclerosis.2017.04.011. Epub 2017 Apr 13.
We aimed at investigating the role of 14q32 microRNAs in intimal hyperplasia and accelerated atherosclerosis; two major contributors to restenosis. Restenosis occurs regularly in patients treated for coronary artery disease and peripheral arterial disease. We have previously shown that inhibition of 14q32 microRNAs leads to increased post-ischemic neovascularization, and microRNA miR-494 also decreased atherosclerosis, while increasing plaque stability. We hypothesized that 14q32 microRNA inhibition has beneficial effects on intimal hyperplasia, as well as accelerated atherosclerosis.
Non-constrictive cuffs were placed around both femoral arteries of C57BL/6J mice to induce intimal hyperplasia. Accelerated atherosclerotic plaque formation was induced in hypercholesterolemic ApoE mice by placing semi-constrictive collars around both carotid arteries. 14q32 microRNAs miR-329, miR-494 and miR-495 were inhibited in vivo using Gene Silencing Oligonucleotides (GSOs).
GSO-495 administration led to a 32% reduction of intimal hyperplasia. Moreover, the number of macrophages in the arterial wall of mice treated with GSO-495 was reduced by 55%. Inhibition of miR-329 and miR-494 had less profound effects on intimal hyperplasia. GSO-495 administration also decreased atherosclerotic plaque formation by 52% and plaques of GSO-495 treated animals showed a more stable phenotype. Finally, cholesterol levels were also decreased in GSO-495 treated animals, via reduction of the VLDL-fraction.
GSO-495 administration decreased our primary outcomes, namely intimal hyperplasia, and accelerated atherosclerosis. GSO-495 administration also favourably affected multiple secondary outcomes, including macrophage influx, plaque stability and total plasma cholesterol levels. We conclude that 14q32 microRNA miR-495 is a promising target for prevention of restenosis.
我们旨在研究14q32微小RNA在内膜增生和动脉粥样硬化加速进程中的作用，这两者是再狭窄的两个主要促成因素。再狭窄在接受冠状动脉疾病和外周动脉疾病治疗的患者中经常发生。我们之前已经表明，抑制14q32微小RNA会导致缺血后新生血管形成增加，微小RNA miR-494也会减少动脉粥样硬化，同时增加斑块稳定性。我们假设抑制14q32微小RNA对内膜增生以及动脉粥样硬化加速进程具有有益作用。
在C57BL/6J小鼠的双侧股动脉周围放置非收缩性袖带以诱导内膜增生。通过在高胆固醇血症的载脂蛋白E（ApoE）小鼠的双侧颈动脉周围放置半收缩性项圈来诱导动脉粥样硬化斑块加速形成。使用基因沉默寡核苷酸（GSO）在体内抑制14q32微小RNA miR-329、miR-494和miR-495。
给予GSO-495可使内膜增生减少32%。此外，用GSO-495处理的小鼠动脉壁中的巨噬细胞数量减少了55%。抑制miR-329和miR-494对内膜增生的影响较小。给予GSO-495还使动脉粥样硬化斑块形成减少52%，并且接受GSO-495处理的动物的斑块表现出更稳定的表型。最后，通过降低极低密度脂蛋白（VLDL）组分，接受GSO-495处理的动物的胆固醇水平也降低了。
给予GSO-495降低了我们的主要结局指标，即内膜增生和动脉粥样硬化加速进程。给予GSO-495还对多个次要结局指标产生了有利影响，包括巨噬细胞流入、斑块稳定性和总血浆胆固醇水平。我们得出结论，14q32微小RNA miR-495是预防再狭窄的一个有前景的靶点。
