A fibronectin fragment alters the metabolism by rabbit intervertebral disc cells in vitro
A fibronectin fragment alters the metabolism by rabbit intervertebral disc cells in vitro
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DOI：
10.1097/01.brs.0000164097.47091.4c
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发表时间：
2005-06-01
期刊：
影响因子：
3
通讯作者：
Balian, G
中科院分区：
文献类型：
--
作者：
Anderson, DG;Li, XD;Balian, G
研究设计.进行了生化和基因表达研究，以确定30 kDa的N-末端纤连蛋白片段（Fn-f）对髓核（NP）外植体培养物中糖胺聚糖含量的影响，以及藻酸盐培养物中NP细胞的基因表达谱。确定Fn-f对藻酸盐培养物和圆盘外植体培养物中NP细胞的影响。椎间盘退变的宏观和组织学特征已被很好地描述，但椎间盘退变的分子生物学仍然知之甚少。虽然纤连蛋白和纤连蛋白片段已知在退行性椎间盘中积累，但纤连蛋白片段在退行性过程中的作用尚未阐明。本研究旨在确定Fn-f对关键基质和降解基因的表达以及对椎间盘基质蛋白的影响。收集新西兰白色兔椎间盘。分离NP细胞并使其在藻酸盐培养物中生长或培养为分离的组织。将培养的细胞暴露于10 nmol/L、100 nmol/L和1 μ mol/L浓度的30-kDa N-末端Fn-f或对照物质，然后进行组织学、生化和基因表达研究。藻酸盐培养的NP细胞保持了与体内椎间盘细胞相似的组织学外观和表型表达模式。这些细胞暴露于Fn-f导致MMP-9、MMP-13和Fas基因的上调以及II型胶原和聚集蛋白聚糖基因的下调。在外植体培养中，与对照相比，Fn-f暴露导致糖胺聚糖含量减少60%。在体外用Fn-f处理NP细胞导致基质蛋白和基因表达的变化，这些变化与体内椎间盘退变过程中观察到的相似。这支持了N-末端纤连蛋白片段在退行性椎间盘疾病中可能的有害作用。
Study Design. A biochemical and gene expression study was conducted to determine the effects of the 30-kDa N-terminal fibronectin fragment (Fn-f) on the glycosaminoglycan content of nucleus pulposus (NP) explant cultures, and on the gene expression profile of NP cells in alginate culture.Objective. To determine the effects of Fn-f on NP cells in alginate culture and disc explant cultures.Summary of Background Data. The macroscopic and histologic features of disc degeneration have been well described, but the molecular biology of disc degeneration remains poorly understood. Although fibronectin and fibronectin fragments are known to accumulate in degenerative discs, the role of fibronectin fragments on the degenerative process has not been elucidated. This study sought to define the effects of Fn-f on the expression of key matrix and degradative genes and on disc matrix proteins.Methods. New Zealand white rabbits discs were harvested. NP cells were either isolated and grown in alginate culture or cultured as explanted tissue. The cultured cells were exposed to 10 nmol/L, 100 nmol/L, and 1 mu mol/L concentrations of 30-kDa N-terminal Fn-f or a control substance and then analyzed histologically, biochemically, and with gene expression studies.Results. Alginate-cultured NP cells maintained a histologic appearance and phenotypic expression pattern similar to disc cells in vivo. Exposure of these cells to Fn-f led to the up-regulation of the MMP-9, MMP-13, and Fas genes and the down-regulation of the Type II collagen and aggrecan genes. In explant culture, Fn-f exposure led to a 60% reduction in glycosaminoglycan content compared with controls.Conclusion. Treatment of NP cells in vitro with Fn-f led to changes in matrix proteins and gene expression similar to those seen during disc degeneration in vivo. This supports a possible detrimental role of the N-terminal fibronectin fragment in degenerative disc disease.