de Gonzalo-Calvo David, Cenarro Ana, Civeira Fernando, Llorente-Cortes Vicenta
Cardiovascular Research Center (CSIC-ICCC), Biomedical Research Institute Sant Pau (IIB Sant Pau), Barcelona, Spain.
Lipid Unit and Molecular Research Laboratory, IIS Aragón, Hospital Universitario Miguel Servet, Zaragoza, Spain.
Clin Investig Arterioscler. 2016 Jul-Aug;28(4):167-77. doi: 10.1016/j.arteri.2016.05.005. Epub 2016 Jun 28.
microRNA (miRNA) expression profile of extracellular vesicles is a potential tool for clinical practice. Despite the key role of vascular smooth muscle cells (VSMC) in cardiovascular pathology, there is limited information about the presence of miRNAs in microparticles secreted by this cell type, including human coronary artery smooth muscle cells (HCASMC). Here, we tested whether HCASMC-derived microparticles contain miRNAs and the value of these miRNAs as biomarkers.
HCASMC and explants from atherosclerotic or non-atherosclerotic areas were obtained from coronary arteries of patients undergoing heart transplant. Plasma samples were collected from: normocholesterolemic controls (N=12) and familial hypercholesterolemia (FH) patients (N=12). Both groups were strictly matched for age, sex and cardiovascular risk factors. Microparticle (0.1-1μm) isolation and characterization was performed using standard techniques. VSMC-enriched miRNAs expression (miR-21-5p, -143-3p, -145-5p, -221-3p and -222-3p) was analyzed using RT-qPCR.
Total RNA isolated from HCASMC-derived microparticles contained small RNAs, including VSMC-enriched miRNAs. Exposition of HCASMC to pathophysiological conditions, such as hypercholesterolemia, induced a decrease in the expression level of miR-143-3p and miR-222-3p in microparticles, not in cells. Expression levels of miR-222-3p were lower in circulating microparticles from FH patients compared to normocholesterolemic controls. Microparticles derived from atherosclerotic plaque areas showed a decreased level of miR-143-3p and miR-222-3p compared to non-atherosclerotic areas.
We demonstrated for the first time that microparticles secreted by HCASMC contain microRNAs. Hypercholesterolemia alters the microRNA profile of HCASMC-derived microparticles. The miRNA signature of HCASMC-derived microparticles is a source of cardiovascular biomarkers.
细胞外囊泡的微小RNA（miRNA）表达谱是一种具有临床应用潜力的工具。尽管血管平滑肌细胞（VSMC）在心血管病理过程中起着关键作用，但关于这种细胞类型分泌的微粒中miRNA的存在情况，包括人冠状动脉平滑肌细胞（HCASMC），相关信息有限。在此，我们测试了HCASMC来源的微粒是否含有miRNA以及这些miRNA作为生物标志物的价值。
从接受心脏移植患者的冠状动脉中获取HCASMC以及来自动脉粥样硬化或非动脉粥样硬化区域的外植体。从以下人群中采集血浆样本：正常胆固醇血症对照组（N = 12）和家族性高胆固醇血症（FH）患者（N = 12）。两组在年龄、性别和心血管危险因素方面进行了严格匹配。使用标准技术进行微粒（0.1 - 1μm）的分离和表征。使用RT-qPCR分析富含VSMC的miRNA表达（miR-21-5p、-143-3p、-145-5p、-221-3p和-222-3p）。
从HCASMC来源的微粒中分离出的总RNA包含小RNA，包括富含VSMC的miRNA。将HCASMC暴露于病理生理条件下，如高胆固醇血症，会导致微粒中miR-143-3p和miR-222-3p的表达水平降低，而细胞中则没有。与正常胆固醇血症对照组相比，FH患者循环微粒中miR-222-3p的表达水平较低。与非动脉粥样硬化区域相比，来自动脉粥样硬化斑块区域的微粒中miR-143-3p和miR-222-3p的水平降低。
我们首次证明HCASMC分泌的微粒含有微小RNA。高胆固醇血症会改变HCASMC来源微粒的微小RNA谱。HCASMC来源微粒的miRNA特征是心血管生物标志物的一个来源。
