From the King's British Heart Foundation Centre, King's College London, London, United Kingdom.
Circ Res. 2013 Oct 25;113(10):1138-47. doi: 10.1161/CIRCRESAHA.113.302400. Epub 2013 Sep 4.
MicroRNAs (miRNAs), in particular miR-29b and miR-30c, have been implicated as important regulators of cardiac fibrosis.
To perform a proteomics comparison of miRNA effects on extracellular matrix secretion by cardiac fibroblasts.
Mouse cardiac fibroblasts were transfected with pre-/anti-miR of miR-29b and miR-30c, and their conditioned medium was analyzed by mass spectrometry. miR-29b targeted a cadre of proteins involved in fibrosis, including multiple collagens, matrix metalloproteinases, and leukemia inhibitory factor, insulin-like growth factor 1, and pentraxin 3, 3 predicted targets of miR-29b. miR-29b also attenuated the cardiac fibroblast response to transforming growth factor-β. In contrast, miR-30c had little effect on extracellular matrix production but opposite effects regarding leukemia inhibitory factor and insulin-like growth factor 1. Both miRNAs indirectly affected cardiac myocytes. On transfection with pre-miR-29b, the conditioned medium of cardiac fibroblasts lost its ability to support adhesion of rat ventricular myocytes and led to a significant reduction of cardiac myocyte proteins (α-actinin, cardiac myosin-binding protein C, and cardiac troponin I). Similarly, cardiomyocytes derived from mouse embryonic stem cells atrophied under pre-miR-29 conditioned medium, whereas pre-miR-30c conditioned medium had a prohypertrophic effect. Levels of miR-29a, miR-29c, and miR-30c, but not miR-29b, were significantly reduced in a mouse model of pathological but not physiological hypertrophy. Treatment with antagomiRs to miR-29b induced excess fibrosis after aortic constriction without overt deterioration in cardiac function.
Our proteomic analysis revealed novel molecular targets of miRNAs that are linked to a fibrogenic cardiac phenotype. Such comprehensive screening methods are essential to define the concerted actions of miRNAs in cardiovascular disease.
MicroRNAs（miRNAs），特别是 miR-29b 和 miR-30c，被认为是心脏纤维化的重要调节因子。
对心脏成纤维细胞细胞外基质分泌的 miRNA 效应进行蛋白质组学比较。
用 miR-29b 和 miR-30c 的前体/反义 miRNA 转染小鼠心脏成纤维细胞，并通过质谱分析其条件培养基。miR-29b 靶向一组与纤维化有关的蛋白，包括多种胶原蛋白、基质金属蛋白酶和白血病抑制因子、胰岛素样生长因子 1 和 pentraxin 3、3 个 miR-29b 的预测靶点。miR-29b 还减弱了心脏成纤维细胞对转化生长因子-β的反应。相比之下，miR-30c 对细胞外基质的产生几乎没有影响，但对白血病抑制因子和胰岛素样生长因子 1 有相反的影响。两种 miRNA 都间接影响心肌细胞。在转染前体 miR-29b 后，心脏成纤维细胞的条件培养基丧失了支持大鼠心室肌细胞黏附的能力，并导致心肌蛋白（α-辅肌动蛋白、心肌肌球蛋白结合蛋白 C 和心肌肌钙蛋白 I）显著减少。同样，在 pre-miR-29 条件培养基中，来自小鼠胚胎干细胞的心肌细胞萎缩，而 pre-miR-30c 条件培养基则具有促肥大作用。在病理性而非生理性肥大的小鼠模型中，miR-29a、miR-29c 和 miR-30c 的水平，但不是 miR-29b，明显降低。用 miR-29b 的拮抗剂治疗主动脉缩窄后会引起过度纤维化，而心脏功能没有明显恶化。
我们的蛋白质组学分析揭示了与纤维化心脏表型相关的 miRNA 的新分子靶标。这种全面的筛选方法对于确定 miRNA 在心血管疾病中的协同作用至关重要。
