Changes in gene expression and protein distribution at different stages of mechanically induced disc degeneration - An in vivo study on the New Zealand white rabbit
Changes in gene expression and protein distribution at different stages of mechanically induced disc degeneration - An in vivo study on the New Zealand white rabbit
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DOI：
10.1002/jor.20055
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发表时间：
2006-03-01
影响因子：
2.8
通讯作者：
Guehring, T
中科院分区：
文献类型：
--
作者：
Omlor, GW;Lorenz, H;Guehring, T
本研究的目的是使用兔模型提高对退行性椎间盘疾病的生物学理解，在该模型中，通过改变施加2.4 MPa外部压缩器械的载荷持续时间来诱导不同阶段的椎间盘退变。基因表达和蛋白质分布进行了分析，在对照组和后1，28，和56天的超生理负荷。为了评估细胞外基质基因，应用定量实时逆转录聚合酶链反应检测I型胶原、II型胶原、双糖蛋白聚糖、核心蛋白聚糖、纤调蛋白、纤连蛋白、聚集蛋白聚糖和骨粘连蛋白。作为分解代谢、抗分解代谢和合成代谢因子的代表，选择基质金属蛋白酶-13（MMP-13）、基质金属蛋白酶组织抑制剂-1（TIMP-1）和骨形态发生蛋白-2（BMP-2）。为了评估蛋白质分布，对胶原蛋白I、胶原蛋白II和BMP-2/4进行免疫组织化学。基质基因表达的特点是两个主要的发展：胶原蛋白I和II，双糖蛋白聚糖，核心蛋白聚糖显示早期升高，随后下调到控制水平，而纤调蛋白，纤连蛋白，聚集蛋白聚糖，骨连接蛋白显示持续上调或保持在类似的水平。MMP-13基因表达的诱导被发现在退变的椎间盘。TIMP-1和BMP-2在高生理负荷后立即升高，并在56天组中呈现最高水平。免疫组化显示压缩后II型胶原和BMP-2/4阳性细胞减少。总之，基质基因表达升高代表了对超生理负荷的早期细胞反应。随着变性的进展，一些miatrix基因增加上调，而其他基因开始下调。分解代谢、抗分解代谢和合成代谢因子的持续上调表明它们在退化过程中的重要作用。(c)2005骨科研究学会。由威利期刊公司出版
The objective of the study was to improve the biological understanding of degenerative disc disease using a rabbit model in which different stages of disc degeneration are induced by variation of the duration of loading with an external compression-device applying 2.4 MPa. Gene expression and protein distribution were analyzed in controls and after 1, 28, and 56 days of hyperphysiologic loading. To evaluate extracellular matrix genes, quantitative real-time reverse-transcriptase polymerase chain reaction was applied for collagen I, collagen II, biglycan, decorin, fibromodulin, fibronectin, aggrecan, and osteonectin. As representatives of catabolic, anticatabolic, and anabolic factors, matrix metalloproteinase-13 (MMP-13), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), and bone morphogenetic protein-2 (BMP-2) were chosen. To evaluate protein distribution, immunohistochemistry was performed for collagen I, collagen II, and BMP-2/4. Matrix gene expression was characterized by two major developments: collagen I and II, biglycan, and decorin showed early elevation followed by later downregulation to control levels, whereas fibromodulin, fibronectin, aggrecan, and osteonectin showed continuous upregulation or remained at similar levels. Induction of MMP-13 gene expression was found in degenerated discs. TIMP-1 and BMP-2 were elevated immediately after hyperphysiologic loading and presented highest levels in the 56-day group. Immunohistochemistry showed less collagen II and BMP-2/4, positive cells after compression. In conclusion, elevated matrix gene expression represents an early cellular response to hyperphysiologic loading. As degeneration progresses, some miatrix genes increase upregulation, whereas other start downregulation. Continuous upregulation of catabolic, anticatabolic, and Anabolic factors indicates their important role in the degeneration process. (c) 2005 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.