MicroRNA-320 regulates matrix metalloproteinase-13 expression in chondrogenesis and interleukin-1β-induced chondrocyte responses
MicroRNA-320 regulates matrix metalloproteinase-13 expression in chondrogenesis and interleukin-1β-induced chondrocyte responses
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MicroRNA-320 调节软骨形成和白细胞介素 1β 诱导的软骨细胞反应中基质金属蛋白酶 13 的表达
DOI：
10.1016/j.joca.2015.12.012
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发表时间：
2016-05-01
影响因子：
7
通讯作者：
Liao, W.
中科院分区：
文献类型：
--
作者：
Meng, F.;Zhang, Z.;Liao, W.
目的：金属蛋白酶（MMP）是骨关节炎（OA）和胶原蛋白降解的关键调节因子，并已被证明参与软骨内骨化。本研究的目的是确定 microRNA-320 (miR-320) 是否在软骨形成和炎症中调节 MMP-13 的表达。 实验设计：在软骨形成的 ATDC5 细胞模型和白细胞介素 1 β (IL-1 β) 处理的原代小鼠软骨细胞 (PMC) 中体外评估 miR-320 表达，并在体内评估正常和 OA 人类软骨中的 miR-320 表达。原位杂交。 ATDC5 和 PMC 分别用 miR-320 或其反义抑制剂 (anti-miR-320) 转染。通过使用特定抑制剂评估激活的 MAP 激酶 (MAPK) 和 NF-κ B 的作用。通过荧光素酶报告基因检测证实了 miR-320 与其 Mmp-13 mRNA 30 非翻译区 (30-UTR) 中的假定结合位点之间的直接相互作用。结果：与正常软骨相比，在软骨形成和肥大 ATDC5 中 miR-320 表达升高，而在 OA 软骨中显着降低。 IL-1 beta 刺激导致 PMC 中 miR-320 表达显着降低。 PMC 和 ATDC5 中 MMP-13 表达的上调与 miR-320 表达的下调相关。在 ATDC5 和 IL-1 β 处理的 PMC 中，miR-320 的过表达抑制了含有 30-UTR 的报告构建体的活性，并抑制了 MMP-13 的表达，而抗 miR-320 的处理则增强了 MMP-13 的表达。 NF-kappa B 和 MAPK 激活下调 miR-320 表达。结论：软骨发育和稳态受 miR-320 影响，miR-320 直接靶向 MMP-13，调节小鼠软骨细胞中的软骨形成和 IL-1 β 刺激的分解代谢效应。 (c) 2016 年国际骨关节炎研究协会。由爱思唯尔有限公司出版。保留所有权利。
Objective: Metalloproteinases (MMPs) are key regulators of osteoarthritis (OA) and collagen degradation and have been shown to participate in endochondral ossification. The aim of this study was to determine whether microRNA-320 (miR-320) regulates the expression of MMP-13 in chondrogenesis and inflammation.Experimental design: miR-320 expression was assessed in vitro, in the ATDC5 cell model of chondrogenesis and in interleukin-1 beta (IL-1 beta)-treated primary mouse chondrocytes (PMCs), and in vivo, in normal and OA human cartilage by in situ hybridization. ATDC5 and PMCs were transfected with miR-320 or its antisense inhibitor (anti-miR-320), respectively. The roles of activated MAP kinases (MAPK) and NF-kappa B were evaluated by using specific inhibitors. Direct interaction between miR-320 and its putative binding site in the 30-untranslated region (30-UTR) of Mmp-13 mRNA was confirmed by the luciferase reporter assay.Results: miR-320 expression was elevated in chondrogenic and hypertrophic ATDC5, while significantly reduced in OA cartilage compared with normal cartilage. Stimulation with IL-1 beta led to a significant reduction in miR-320 expression in PMCs. Upregulation of MMP-13 expression was correlated with downregulation of miR-320 expression in both PMCs and ATDC5. Overexpression of miR-320 suppressed the activity of a reporter construct containing the 30-UTR and inhibited MMP-13 expression in both ATDC5 and IL-1 beta -treated PMCs, while treatment with anti-miR-320 enhanced MMP-13 expression. NF-kappa B and MAPK activation downregulated miR-320 expression.Conclusion: Cartilage development and homeostasis are influenced by miR-320, which directly targets MMP-13 and regulates chondrogenesis and the IL-1 beta-stimulated catabolic effect in mouse chondrocytes. (c) 2016 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.