Department of Orthopaedic Surgery, Carolinas Medical Center, PO Box 32861, Charlotte, NC 28232, USA.
Exp Mol Pathol. 2011 Oct;91(2):534-9. doi: 10.1016/j.yexmp.2011.06.001. Epub 2011 Jun 12.
During disk degeneration, annulus dehydration and matrix fraying culminate in the formation of tears through which nucleus and annulus disk material may rupture, causing radicular pain. Annular tears are present in more than half of the patients in early adulthood and are almost always present in the elderly. Aggrecan, which provides the disk with a shock absorber function under loading, is a key disk extracellular matrix (ECM) component. The objective of the present study was to assess the immunolocalization of aggrecan in the annulus, and to assess molecular gene expression patterns in the annulus ECM utilizing microarray analysis. Immunohistochemistry was performed on 45 specimens using an anti-human aggrecan antibody. Affymetrix microarray gene expression studies used the extracellular matrix ontology approach to evaluate an additional 6 grade I-II, 9 grade III, and 4 grade IV disks. Grade III/IV disks were compared to healthier grade I/II disks. Healthy and less degenerated disks showed a general uniform aggrecan immunolocalization; more degenerated disks contained regions with little or no identifiable aggrecan localization. In degenerated disks, molecular studies showed a significant downregulation of aggrecan, ADAMTS-like 3, and ADAMTS10. Collagen types III and VIII, fibronectin, decorin, connective tissue growth factor, TIMP-3, latent TGF-β binding protein 2 and TGF-β1 were significantly upregulated with fold changes ranging from 2.4 to 9.8. Findings here help us better understand changes in the immunohistochemical distribution of a key proteoglycan during disk aging. Such information may have application as we work towards biologic therapies to improve the aging/degenerating disk matrix.
在椎间盘退变过程中,纤维环脱水和基质磨损最终导致撕裂的形成,核和纤维环的椎间盘物质可能会通过这些撕裂破裂,引起神经根疼痛。在早期成年患者中,超过一半的患者存在环状撕裂,而在老年人中几乎总是存在。聚集蛋白聚糖(aggrecan)为椎间盘提供了在加载下的减震器功能,是椎间盘细胞外基质(ECM)的关键成分。本研究的目的是评估聚集蛋白聚糖在纤维环中的免疫定位,并利用微阵列分析评估纤维环 ECM 的分子基因表达模式。使用抗人聚集蛋白聚糖抗体对 45 个标本进行了免疫组织化学染色。Affymetrix 微阵列基因表达研究采用细胞外基质本体论方法,对另外 6 个 I-II 级、9 个 III 级和 4 个 IV 级椎间盘进行了评估。III/IV 级椎间盘与更健康的 I/II 级椎间盘进行了比较。健康和退变程度较低的椎间盘显示出一般均匀的聚集蛋白聚糖免疫定位;退变程度较高的椎间盘含有聚集蛋白聚糖定位较少或无法识别的区域。在退变的椎间盘,分子研究显示聚集蛋白聚糖、ADAMTS 样 3 和 ADAMTS10 的表达显著下调。胶原类型 III 和 VIII、纤维连接蛋白、饰胶蛋白、结缔组织生长因子、TIMP-3、潜伏 TGF-β结合蛋白 2 和 TGF-β1 的表达显著上调,倍数变化范围为 2.4 至 9.8。这些发现有助于我们更好地了解在椎间盘老化过程中关键蛋白聚糖的免疫组织化学分布变化。这些信息可能在我们努力开发改善老化/退变椎间盘基质的生物治疗方法时具有应用价值。