Injectable hyaluronic acid down-regulates interferon signaling molecules, IGFBP3 and IFIT3 in the bovine intervertebral disc
Injectable hyaluronic acid down-regulates interferon signaling molecules, IGFBP3 and IFIT3 in the bovine intervertebral disc
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DOI：
10.1016/j.actbio.2016.12.029
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发表时间：
2017-04-01
影响因子：
9.7
通讯作者：
Pandit, Abhay
中科院分区：
文献类型：
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作者：
Kazezian, Zepur;Li, Zhen;Pandit, Abhay
腰痛是20至50岁人群致残的主要原因，造成了严重的社会经济负担。目前再生医学的重点是识别分子标记，以促进靶向治疗的设计。在此之前，我们已经证明，在人纤维环（AF）中，抗增殖干扰素诱导的四肽重复3蛋白（IFIT3）和促凋亡胰岛素样生长因子结合蛋白-3 （IGFBP3）作为炎症细胞因子干扰素α (IFN α)信号通路的下游靶点被上调。在这里，我们假设注射透明质酸（HA）对损伤和IFN α (2) β炎症的牛椎间盘（IVD）具有抗炎和基质调节作用。在牛IVD器官培养模型中，采用IFN α (2) β刺激的AF缺损椎间盘，检测HA对IFN α (2) β通路以及基质蛋白聚集蛋白和胶原i的影响，采用qRT-PCR方法评估IFN α (2) β信号分子的基因表达。此外，免疫染色法检测蛋白表达。我们的研究结果显示，HA处理显著下调炎症组IFNARI、IFNAR2、STAT1/2、JAK1、IFIT3和IGFBP3 mRNA的表达。蛋白分析证实了PCR结果。在细胞外基质中，HA治疗损伤和炎症椎间盘后，aggrecan和collagen I上调，ADAMTS4下调。因此，HA既具有抗炎作用，导致AF中IFIT3和IGFBP3的下调，又具有基质调节作用，通过上调聚集蛋白和I型胶原的表达。意义声明退行性IVD的促炎环境对再生治疗提出了挑战。研究表明，透明质酸通过下调IFNARI和IFNAR2、信号分子STAT1、STAT2、JAK1以及下游凋亡靶点IGFBP3和IFIT3发挥抗炎分子的作用。我们还证明了透明质酸通过增加聚集蛋白和胶原I的合成以及下调炎症条件下降解基质的ADAMTS4来调节椎间盘基质环境。这项工作的意义在于，透明质酸作为一种抗炎分子，通过将椎间盘环境转向更合成代谢的状态，并通过促进天然IVD基质的产生。(C) 2016材料学报Elsevier Ltd.出版。版权所有。
Low back pain which is a major cause of disability for people aged between 20 and 50 years imposes a serious socio-economic burden. The current focus of regenerative medicine is on identifying molecular markers to facilitate the design of targeted therapeutics. Previously, we have demonstrated that expression of the anti-proliferative interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and proapoptotic insulin-like growth factor-binding protein-3 (IGFBP3), are up-regulated as downstream targets of the inflammatory cytokine interferon alpha (IFN alpha) signaling pathway in the human annulus fibrosus (AF). Here, we hypothesised that injection of hyaluronic acid (HA) would have an anti-inflammatory and matrix modulatory effect on injured and IFN alpha(2)beta inflamed bovine intervertebral discs (IVD). Discs with an AF defect and challenged with IFN alpha(2)beta were used in a bovine IVD organ culture model to test the effect of HA on the IFN alpha(2)beta pathway, as well as the matrix proteins aggrecan and collagen I. qRT-PCR was used to assess the gene expression of IFN alpha(2)beta signaling molecules. Additionally, immunostaining was used to measure protein expression. Our results show that HA treatment significantly down-regulates IFNARI, IFNAR2, STAT1/2, JAK1, IFIT3 and IGFBP3 mRNA expression in the inflamed groups. Protein analysis confirmed the PCR results. In the extracellular matrix, aggrecan and collagen I were up-regulated while ADAMTS4 was down-regulated upon treatment of the injured and inflamed discs with HA. Hence, HA demonstrates both an anti-inflammatory role, resulting in the down-regulation of IFIT3 and IGFBP3 in the AF, and a matrix modulatory effect by up-regulating aggrecan and collagen I expression.Statement of SignificanceThe pro-inflammatory environment of the degenerated IVD represents a challenge for regenerative therapies. The study demonstrates that hyaluronan acts as an anti-inflammatory molecule by down regulating IFNARI and IFNAR2, the signaling molecules STAT1, STAT2,JAK1 and the downstream apoptotic targets IGFBP3 and IFIT3. We also demonstrated that hyaluronan modulates the disc matrix environment by increasing aggrecan and collagen I synthesis and down-regulating ADAMTS4 that degrades the matrix under inflammatory conditions. The significance of this work lies in the fact that hyaluronan acts as an anti-inflammatory molecule by shifting the disc environment towards a more anabolic state and by promoting native IVD matrix production. (C) 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.