Yuan Y, Zhang G Q, Chai W, Ni M, Xu C, Chen J Y
Department of Orthopaedics, Chinese PLA General Hospital, No.28 Fuxing Road,Haidian District,Beijing 100853,China and, Jinan Military General Hospital, No.25, Shifan Road, Tianqiao District, Jinan 250031, Shandong, China.
Department of Orthopaedics, Chinese PLA General Hospital, General Hospital, No.28 Fuxing Road, Haidian District, Beijing 100853, China.
Bone Joint Res. 2016 Oct;5(10):523-530. doi: 10.1302/2046-3758.510.BJR-2016-0074.R2.
Osteoarthritis (OA) is characterised by articular cartilage degradation. MicroRNAs (miRNAs) have been identified in the development of OA. The purpose of our study was to explore the functional role and underlying mechanism of miR-138-5p in interleukin-1 beta (IL-1β)-induced extracellular matrix (ECM) degradation of OA cartilage.
Human articular cartilage was obtained from patients with and without OA, and chondrocytes were isolated and stimulated by IL-1β. The expression levels of miR-138-5p in cartilage and chondrocytes were both determined. After transfection with miR-138-5p mimics, allele-specific oligonucleotide (ASO)-miR-138-5p, or their negative controls, the messenger RNA (mRNA) levels of aggrecan (ACAN), collagen type II and alpha 1 (COL2A1), the protein levels of glycosaminoglycans (GAGs), and both the mRNA and protein levels of matrix metalloproteinase (MMP)-13 were evaluated. Luciferase reporter assay, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot were performed to explore whether Forkhead Box C1 (FOCX1) was a target of miR-138-5p. Further, we co-transfected OA chondrocytes with miR-138-5p mimics and pcDNA3.1 (+)-FOXC1 and then stimulated with IL-1β to determine whether miR-138-5p-mediated IL-1β-induced cartilage matrix degradation resulted from targeting FOXC1.
MiR-138-5p was significantly increased in OA cartilage and in chondrocytes in response to IL-1β-stimulation. Overexpression of miR-138-5p significantly increased the IL-1β-induced downregulation of COL2A1, ACAN, and GAGs, and increased the IL-1β-induced over expression of MMP-13.We found that FOXC1 is directly regulated by miR-138-5p. Additionally, co-transfection with miR-138-5p mimics and pcDNA3.1 (+)-FOXC1 resulted in higher levels of COL2A1, ACAN, and GAGs, but lower levels of MMP-13.
miR-138-5p promotes IL-1β-induced cartilage degradation in human chondrocytes, possibly by targeting FOXC1.Cite this article: Y. Yuan, G. Q. Zhang, W. Chai,M. Ni, C. Xu, J. Y. Chen. Silencing of microRNA-138-5p promotes IL-1β-induced cartilage degradation in human chondrocytes by targeting FOXC1: miR-138 promotes cartilage degradation. Bone Joint Res 2016;5:523-530. DOI: 10.1302/2046-3758.510.BJR-2016-0074.R2.
骨关节炎（OA）的特征是关节软骨降解。微小RNA（miRNA）已被证实在OA的发展过程中发挥作用。本研究旨在探讨miR-138-5p在白细胞介素-1β（IL-1β）诱导的OA软骨细胞外基质（ECM）降解中的功能作用及潜在机制。
从患OA和未患OA的患者获取人关节软骨，分离软骨细胞并用IL-1β刺激。测定软骨和软骨细胞中miR-138-5p的表达水平。用miR-138-5p模拟物、等位基因特异性寡核苷酸（ASO）-miR-138-5p或其阴性对照转染后，评估聚集蛋白聚糖（ACAN）、Ⅱ型胶原蛋白α1（COL2A1）的信使核糖核酸（mRNA）水平、糖胺聚糖（GAGs）的蛋白水平以及基质金属蛋白酶（MMP）-13的mRNA和蛋白水平。进行荧光素酶报告基因检测、定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法，以探究叉头框C1（FOXC1）是否为miR-138-5p的靶标。此外，我们将miR-138-5p模拟物与pcDNA3.1（+）-FOXC1共转染OA软骨细胞，然后用IL-1β刺激，以确定miR-138-5p介导的IL-1β诱导的软骨基质降解是否是通过靶向FOXC1所致。
在OA软骨以及经IL-1β刺激的软骨细胞中，miR-138-5p显著上调。miR-138-5p过表达显著增加了IL-1β诱导的COL2A1、ACAN和GAGs的下调，并增加了IL-1β诱导的MMP-13的过表达。我们发现FOXC1直接受miR-138-5p调控。此外，miR-138-5p模拟物与pcDNA3.1（+）-FOXC1共转染导致COL2A1、ACAN和GAGs水平升高，但MMP-13水平降低。
miR-138-5p可能通过靶向FOXC1促进IL-1β诱导的人软骨细胞软骨降解。引用本文：Y. Yuan, G. Q. Zhang, W. Chai, M. Ni, C. Xu, J. Y. Chen. 微小RNA-138-5p沉默通过靶向FOXC1促进IL-1β诱导的人软骨细胞软骨降解：miR-138促进软骨降解。骨关节研究2016；5：523 - 530。DOI：10.1302/2046 - 3758.510.BJR - 2016 - 0074.R2。
