miR-130b-3p Modulates Epithelial-Mesenchymal Crosstalk in Lung Fibrosis by Targeting IGF-1.
miR-130b-3p Modulates Epithelial-Mesenchymal Crosstalk in Lung Fibrosis by Targeting IGF-1.
复制标题
miR-130b-3p 通过靶向 IGF-1 调节肺纤维化中的上皮-间质串扰
DOI：
10.1371/journal.pone.0150418
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Dai H
中科院分区：
文献类型：
--
作者：
Li S;Geng J;Xu X;Huang X;Leng D;Jiang D;Liang J;Wang C;Jiang D;Dai H
特发性肺纤维化（IPF）是一种慢性、进行性和通常致死性的纤维化肺病，病因和发病机制基本未知。有证据表明microRNA（miRNA）有助于IPF的发病机制。在这项研究中，我们试图鉴定miRNA表达特征并确定miR-130 b-3 p在肺纤维化中的作用。通过Affyphin微阵列测定IPF患者和正常供体肺的miRNA表达谱，并用Affyphin微阵列测定转录组。通过生物信息学分析，建立了其功能、信号通路和miRNA-mRNA网络。用荧光素酶和ELISA方法对目的基因进行鉴定。使用共培养系统评估miRNA转染的上皮对成纤维细胞活性的影响。通过qRT-PCR、蛋白质印迹、Transwell和BrdU测定来确定成纤维细胞活性。在IPF肺中，7种miRNA显著减少，其中miR-130 b-3 p在miRNA-mRNA网络中最高。胰岛素样生长因子（IGF-1）是miR-130 b-3 p在上皮中的靶基因。在共培养体系中，抑制上皮细胞中的miR-130 b-3 p可诱导成纤维细胞表达I型胶原，并增强成纤维细胞的增殖和迁移能力，模拟外源性IGF-1对成纤维细胞的作用。用抗体中和IGF-1显著降低了miR-130 b-3 p转染上皮细胞对成纤维细胞活化的调节作用。我们的研究结果表明，miR-130 b-3 p在IPF肺中下调。miR-130 b-3 p下调通过促进肺上皮细胞分泌IGF-1而促进成纤维细胞的活化和上皮-间充质串扰的失调，表明该miRNA在预防肺纤维化中的关键调节作用。
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive and usually lethal fibrotic lung disease with largely unknown etiology and pathogenesis. Evidence suggests microRNAs (miRNA) contribute to pathogenesis of IPF. In this study, we sought to identify miRNA expression signatures and determine the role of miR-130b-3p in lung fibrosis. The miRNA expression profile of the lungs from patients with IPF and normal donors was determined by Affymetrix microarray, and transcriptome with Affymetrix array. The functions and signal pathways as well as miRNA-mRNA networks were established by bioinformatics analysis. Luciferase assays and ELISA were used to confirm the miRNA target gene. The effect of miRNA-transfected epithelium on fibroblast activities was assessed using a co-culture system. The fibroblast activities were determined by qRT-PCR, western blotting, Transwell and BrdU assays. Seven miRNAs were significantly decreased in IPF lungs, with miR-130b-3p being the highest in the miRNA-mRNA network. Insulin-like growth factor (IGF-1) was a target gene of miR-130b-3p in the epithelium. miR-130b-3p inhibition in the epithelium induced collagen I expression and enhanced the proliferation and migration ability of fibroblast in co-culture systems, which mimicked the functions of exogenous IGF-1 on fibroblasts. Neutralizing IGF-1 with an antibody significantly reduced the modulatory effects of miR-130b-3p inhibitor-transfected epithelium on the activation of fibroblasts. Our results show that miR-130b-3p was downregulated in IPF lungs. miR-130b-3p downregulation contributed to the activation of fibroblasts and the dysregulated epithelial-mesenchymal crosstalk by promoting IGF-1 secretion from lung epithelium, suggesting a key regulatory role for this miRNA in preventing lung fibrosis.