| 徐敏铭,温子欢,官伟,王绪君,黄常盛.SOX-9通过调控ADAMTS保护椎间盘[J].脊柱外科杂志,2019,17(6):425-430. |
SOX-9通过调控ADAMTS保护椎间盘  点此下载全文 (Fulltext) |
| 徐敏铭1△ 温子欢2△ 官伟1 王绪君1 黄常盛3* |
1. 上饶惠阳医院骨科, 上饶 334000;
2. 上饶市立医院骨科, 上饶 334000;
3. 上饶清水医院骨科, 上饶 334000 |
| 基金项目： |
| DOI：10.3969/j.issn.1672-2957.2019.06.012 |
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| 摘要： |
| 目的 探讨转录因子SOX-9和含Ⅰ型血小板反应蛋白的解聚蛋白样金属蛋白酶（ADAMTS）对椎间盘退行性变的影响及二者之间的关系，为进一步理解椎间盘退行性变发生机制提供实验依据。方法 收集不同退行性变等级的椎间盘组织，通过实时荧光定量PCR、免疫组织化学染色和蛋白质印迹分析等方法检测椎间盘组织中SOX-9、DAMTS-4、ADAMTS-5的表达。原代培养正常椎间盘中的髓核细胞，通过转染小干扰RNA（siRNA）干扰SOX-9、ADAMTS-4、ADAMTS-5的表达，采用实时荧光定量PCR和蛋白质印迹分析检测细胞外基质成分——Ⅱ型胶原（COL2A1）、蛋白聚糖（ACAN）、软骨寡聚物基质蛋白（COMP）的表达变化，通过荧光素酶实验评估SOX-9对ADAMTS-4、ADAMTS-5启动子活性的影响。结果 与正常椎间盘组织相比，发生退行性变椎间盘组织中SOX-9表达降低，而ADAMTS-4、ADAMTS-5表达升高。单纯干扰髓核细胞中SOX-9的表达，COL2A1、ACAN及COMP表达均下调，ADAMTS-4、ADAMTS-5表达上调；同时干扰SOX-9、ADAMTS-4及ADAMTS-5表达后，COL2A1、ACAN及COMP表达较单纯干扰SOX-9表达明显升高。荧光素酶实验结果显示SOX-9可显著下调ADAMTS-4、ADAMTS-5启动子活性，提示二者具有直接调控关系。结论 SOX-9通过直接调控ADAMTS-4、ADAMTS-5表达进而影响髓核细胞外基质成分的表达，最终发挥保护椎间盘的作用。 |
| 关键词：转录因子 ADAM蛋白质类 基因表达调控 椎间盘退行性变 |
| SOX-9 protect intervertebral disc from degeneration through ADAMTS Fulltext |
| XU Min-ming1△ WEN Zi-huan2△ GUAN Wei1 WANG Xu-jun1 HUANG Chang-sheng3* |
1. Department of Orthopaedics, Shangrao Huiyang Hospital, Shangrao 334000, Jiangxi, China;
2. Department of Orthopaedics, Shangrao Municipal Hospital, Shangrao 334000, Jiangxi, China;
3. Department of Orthopaedics, Shangrao Qingshui Hospital, Shangrao 334000, Jiangxi, China |
| Fund Project: |
| Abstract: |
| Objective To investigate the relationship between transcription factor SOX-9 and ADAMTS(A disintergrin like and metalloproteinase with thrombospondin typeⅠmotifs) in intervertebral discs and their effects on disc degeneration,to provide a theoretical basis for further understanding the pathogenesis of intervertebral disc degeneration. Methods The expressions of SOX-9 and ADAMTS in different degenerative discs were detected by real-time fluorescence quantitative PCR,immunohistochemistry and Western blotting. The nucleus pulposus cells in normal intervertebral discs were extracted for primary culture,and SOX-9 as well as ADAMTS were interfered by transfection of small interfering RNA(siRNA). The expressions of extracellular matrix components,including collagenⅡ(COL2A1),aggrecan(ACAN) and cartilage oligomeric matrix protein(COMP),were detected by real-time PCR and Western blotting. Finally,the effect of SOX-9 on the activity of ADAMTS promoters were evaluated by Luciferase assay. Results Compared with normal intervertebral discs,the expression of SOX-9 was decreased and ADAMTS were increased(P<0.05) in degenerated discs. The expressions of COL2A1,ACAN and COMP were decreased and ADAMTS expression increased with SOX-9 single interference,however,the expression of COL2A1,ACAN and COMP were significantly increased with SOX-9 and ADAMTS co-interference compared with SOX-9 single interference. Conclusion SOX-9 affects the expression of extracellular matrix components in the nucleus pulposus cells by directly regulating the expression of ADAMTS,and ultimately plays a role in protecting the intervertebral disc. |
| Keywords:Transcription factors ADAM proteins Gene expression regulation Intervertebral disc degeneration |
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