Center of Molecular Biology and Pharmacogenetics, Scientific and Technological Bioresource Nucleus (BIOREN), Universidad de La Frontera, Av. Francisco Salazar, 01145 Temuco, Chile.
Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Sciences, Universidade de São Paulo, Av. Professor Lineu Prestes 580, São Paulo, Brazil.
Arch Biochem Biophys. 2014 Sep 1;557:28-35. doi: 10.1016/j.abb.2014.04.009. Epub 2014 Apr 30.
New vessel formation plays a critical role in the progression and vulnerability of atherosclerotic lesions. It has been shown that polyphenols from propolis attenuate the progression of atherosclerosis and also exert inhibitory effects on angiogenic factors. However, the mechanisms underlying these effects are not completely understood. Thus, this study aimed to identify microRNAs (miRNAs) involved in the modulation of pro-angiogenic factors in the atherosclerotic plaques of LDL receptor gene knockout mice treated with a polyphenol-rich extract of Chilean propolis. The progression of the atherosclerotic lesions was significantly attenuated in treated mice compared with control mice. Using microarray analysis and a bioinformatic approach, we identified 29 differentially expressed miRNAs. Many of these miRNAs were involved in biological processes associated with angiogenesis, such as the cell cycle, cell migration, cell growth and proliferation. Among them, three miRNAs (miR-181a, miR-106a and miR-20b) were over-expressed and inversely related to the expression of Vegfa (vascular endothelial growth factor A) and Hif1a (hypoxia inducible factor 1 alpha). In addition, VEGF-A protein expression was attenuated in histological sections obtained from the aortic sinuses of treated mice. VEGFA is a key pro-angiogenic factor in atherosclerotic plaques, and Hif1a, which is expressed in the necrotic nucleus of the atheroma, is its main inducer. We found a correlation between the over-expression of miR-181a, miR-106a and miR-20b and their target genes, Hif1a and Vegfa, which is consistent with attenuation of the atherosclerotic lesion. In conclusion, our data analysis provides evidence that the anti-angiogenic effects of polyphenols from Chilean propolis can be modulated by miRNAs, in particular miR-181a, miR-106a and miR-20b.
新血管形成在动脉粥样硬化病变的进展和易损性中起着关键作用。已经表明，来自蜂胶的多酚可减轻动脉粥样硬化的进展，并对血管生成因子发挥抑制作用。然而，这些作用的机制尚不完全清楚。因此，本研究旨在确定与 LDL 受体基因敲除小鼠动脉粥样硬化斑块中促血管生成因子调节相关的 microRNAs（miRNAs），这些小鼠用富含多酚的智利蜂胶提取物处理。与对照组相比，治疗组小鼠的动脉粥样硬化病变进展明显减轻。通过微阵列分析和生物信息学方法，我们鉴定出 29 个差异表达的 miRNAs。其中许多 miRNA 参与与血管生成相关的生物学过程，例如细胞周期、细胞迁移、细胞生长和增殖。其中，三个 miRNAs（miR-181a、miR-106a 和 miR-20b）过表达，与 Vegfa（血管内皮生长因子 A）和 Hif1a（缺氧诱导因子 1α）的表达呈负相关。此外，治疗组小鼠主动脉窦组织切片中的 VEGF-A 蛋白表达减弱。VEGFA 是动脉粥样硬化斑块中的关键促血管生成因子，而在动脉粥样硬化坏死核中表达的 Hif1a 是其主要诱导因子。我们发现 miR-181a、miR-106a 和 miR-20b 的过表达与其靶基因 Hif1a 和 Vegfa 之间存在相关性，这与动脉粥样硬化病变的减轻一致。总之，我们的数据分析提供了证据，表明智利蜂胶中的多酚的抗血管生成作用可以通过 miRNAs 来调节，特别是 miR-181a、miR-106a 和 miR-20b。
