MicroRNA-7 regulates IL-1β-induced extracellular matrix degeneration by targeting GDF5 in human nucleus pulposus cells
MicroRNA-7 regulates IL-1β-induced extracellular matrix degeneration by targeting GDF5 in human nucleus pulposus cells
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DOI：
10.1016/j.biopha.2016.08.062
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发表时间：
2016-10-01
影响因子：
7.5
通讯作者：
Yang, Cao
中科院分区：
文献类型：
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作者：
Liu, Wei;Zhang, Yukun;Yang, Cao
白细胞介素-1 β（IL-1 β）诱导的细胞外基质变性在椎间盘退变（IDD）发病机制中的确切作用目前尚不清楚。最近的证据表明，微小RNA（miRNAs）与IDD相关，但它们在髓核（NP）组织细胞外基质降解中的功能也知之甚少。本研究的目的是评估miR-7在IL-1 β诱导的椎间盘退变中的表达和功能作用。使用定量逆转录-聚合酶链反应扩增分析研究了退行性NP组织和IL-1 β诱导的NP细胞中miR-7的表达水平。然后利用双荧光素酶报告基因测定来确定生长分化因子5（GDF 5）是否是miR-7的靶标。最后，确定已知基质组分和基质降解酶的mRNA和蛋白水平，以阐明miR-7在IL-1 β诱导的椎间盘退变中的功能。在这项研究中，我们发现miR-7在人类退行性NP组织和IL-1 β刺激的NP细胞中高度表达，与正常对照相比。我们还确定GDF 5是miR-7的靶点。功能分析显示，miR-7的过表达显著增强了IL-1 β诱导的细胞外基质变性，而miR-7功能的抑制阻止了NP细胞响应于IL-1 β的有害分解代谢变化。此外，通过miR-7沉默对IL-1 β诱导的NP细胞外基质变性的预防被GDF 5 siRNA减弱。这些发现表明，miR-7通过靶向GDF 5而导致椎间盘中ECM受损，因此miR-7可能代表预防IDD的新治疗靶点。(C)2016 Elsevier Masson SAS。All rights reserved.
The precise role of interleukin-1 beta (IL-1 beta)-induced extracellular matrix degeneration in the pathogenesis of intervertebral disc degeneration (IDD) is currently unknown. Recent evidence has revealed that microRNAs (miRNAs) are associated with IDD, but their function in the extracellular matrix degradation of nucleus pulposus (NP) tissues is also poorly understood. The aim of this study was to evaluate the expression and functional role of miR-7 in IL-1 beta-induced disc degeneration. The expression level of miR-7 was investigated in degenerative NP tissues and in IL-1 beta-induced NP cells using quantitative reverse transcription-polymerase chain reaction amplification analysis. A dual-luciferase reporter assay was then utilized to determine whether growth differentiation factor 5 (GDF5) is a target of miR-7. Finally, mRNA and protein levels of known matrix components and of matrix degradation enzymes were determined to elucidate the function of miR-7 in IL-1 beta-induced disc degeneration. In this study, we found that miR-7 is highly expressed in human degenerative NP tissues and in IL-1 beta stimulated NP cells compared to normal controls. We also determined that GDF5 was a target of miR-7. Functional analysis showed that the overexpression of miR-7 significantly enhanced the IL-1 beta-induced extracellular matrix degeneration, whereas inhibition of miR-7 function by antagomiR-7 prevented NP cell detrimental catabolic changes in response to IL-1 beta. Additionally, the prevention of IL-1 beta-induced NP extracellular matrix degeneration by miR-7 silencing was attenuated by GDF5 siRNA. These findings suggest that miR-7 contributes to an impaired ECM in intervertebral discs through targeting GDF5 and miR-7 might therefore represent a novel therapeutic target for the prevention of IDD. (C) 2016 Elsevier Masson SAS. All rights reserved.