Transcriptional profiling and biochemical analysis of mechanically induced cartilaginous tissues in a rat model.
Transcriptional profiling and biochemical analysis of mechanically induced cartilaginous tissues in a rat model.
复制标题
大鼠模型中机械诱导的软骨组织的转录谱和生化分析。
DOI：
10.1002/art.27343
复制
发表时间：
2010
影响因子：
--
通讯作者：
Morgan,EliseF
中科院分区：
文献类型：
--
作者：
SalisburyPalomares,KristyT;Gerstenfeld,LouisC;Wigner,NathanA;Lenburg,MarcE;Einhorn,ThomasA;Morgan,EliseF
目的通过分析机械诱导软骨形成过程中信使RNA的表达，表征出生后体内软骨形成的分子表达模式。方法采用非临界尺寸的股骨横截骨术对退休的繁殖大鼠进行手术。实验动物（n = 45）于术后第10天开始对截骨间隙进行弯曲刺激（矢状面60°循环运动15分钟/天）。对照组动物（n = 32）连续进行刚性固定。在手术后第10、17、24和38天分离的信使RNA使用包含608个基因的微阵列进行分析，这些基因涉及骨骼发育、组织分化、骨折愈合和机械转导。将受刺激组织中的糖胺聚糖（GAG）含量与天然关节软骨中的含量进行比较，作为评估组织软骨发育进展的一种手段。结果100个差异表达基因中大部分在机械刺激下表达上调。这些基因中有许多与关节软骨发育和维持、关节发育、细胞粘附、细胞外基质合成、信号转导和骨骼发育有关。定量实时聚合酶链反应结果与微阵列结果一致。受刺激组织的GAG含量随着时间的推移而增加，与术后第38天关节软骨的GAG含量没有差异。我们的研究结果表明，机械刺激导致主要参与关节腔形态发生和关节软骨功能的基因上调。对这种刺激的进一步研究可能会确定出生后透明软骨形成所需的关键信号事件。
ObjectiveTo characterize patterns of molecular expression that lead to cartilage formation in vivo in a postnatal setting, by profiling messenger RNA expression across the time course of mechanically induced chondrogenesis.MethodsRetired breeder Sprague‐Dawley rats underwent a noncritical‐sized transverse femoral osteotomy. Experimental animals (n = 45) were subjected to bending stimulation (60° cyclic motion in the sagittal plane for 15 minutes/day) of the osteotomy gap beginning on day 10 after the operation. Control animals (n = 32) experienced continuous rigid fixation. Messenger RNA isolated on days 10, 17, 24, and 38 after surgery was analyzed using a microarray containing 608 genes involved in skeletal development, tissue differentiation, fracture healing, and mechanotransduction. The glycosaminoglycan (GAG) content in the stimulated tissues was compared with that in native articular cartilage as a means of assessing the progression of chondrogenic development of the tissues.ResultsThe majority of the 100 genes that were differentially expressed were up‐regulated in response to mechanical stimulation. Many of these genes are associated with articular cartilage development and maintenance, diarthrodial joint development, cell adhesion, extracellular matrix synthesis, signal transduction, and skeletal development. Quantitative real‐time polymerase chain reaction results were consistent with the microarray findings. The GAG content of the stimulated tissues increased over time and was no different from that of articular cartilage on day 38 after surgery.ConclusionOur findings indicate that mechanical stimulation causes up‐regulation of genes that are principally involved in joint cavity morphogenesis and critical to articular cartilage function. Further study of this type of stimulation may identify key signaling events required for postnatal hyaline cartilage formation.