Rioja Inmaculada, Clayton Chris L, Graham Simon J, Life Paul F, Dickson Marion C
Rheumatoid Arthritis Disease Biology Department, GlaxoSmithKline, Medicines Research Centre, Stevenage, UK.
Arthritis Res Ther. 2005;7(1):R101-17. doi: 10.1186/ar1458. Epub 2004 Nov 19.
Experimental arthritis models are considered valuable tools for delineating mechanisms of inflammation and autoimmune phenomena. Use of microarray-based methods represents a new and challenging approach that allows molecular dissection of complex autoimmune diseases such as arthritis. In order to characterize the temporal gene expression profile in joints from the reactivation model of streptococcal cell wall (SCW)-induced arthritis in Lewis (LEW/N) rats, total RNA was extracted from ankle joints from naive, SCW injected, or phosphate buffered saline injected animals (time course study) and gene expression was analyzed using Affymetrix oligonucleotide microarray technology (RAE230A). After normalization and statistical analysis of data, 631 differentially expressed genes were sorted into clusters based on their levels and kinetics of expression using Spotfire profile search and K-mean cluster analysis. Microarray-based data for a subset of genes were validated using real-time PCR TaqMan analysis. Analysis of the microarray data identified 631 genes (441 upregulated and 190 downregulated) that were differentially expressed (Delta > 1.8, P < 0.01), showing specific levels and patterns of gene expression. The genes exhibiting the highest fold increase in expression on days -13.8, -13, or 3 were involved in chemotaxis, inflammatory response, cell adhesion and extracellular matrix remodelling. Transcriptome analysis identified 10 upregulated genes (Delta > 5), which have not previously been associated with arthritis pathology and are located in genomic regions associated with autoimmune disease. The majority of the downregulated genes were associated with metabolism, transport and regulation of muscle development. In conclusion, the present study describes the temporal expression of multiple disease-associated genes with potential pathophysiological roles in the reactivation model of SCW-induced arthritis in Lewis (LEW/N) rat. These findings improve our understanding of the molecular events that underlie the pathology in this animal model, which is potentially a valuable comparator to human rheumatoid arthritis (RA).
实验性关节炎模型被认为是阐明炎症和自身免疫现象机制的有价值工具。基于微阵列的方法的应用代表了一种新的且具有挑战性的途径,它能够对诸如关节炎等复杂自身免疫性疾病进行分子剖析。为了描绘Lewis(LEW/N)大鼠中链球菌细胞壁(SCW)诱导的关节炎再激活模型关节中的时间基因表达谱,从未处理、注射SCW或注射磷酸盐缓冲盐水的动物的踝关节中提取总RNA(时间进程研究),并使用Affymetrix寡核苷酸微阵列技术(RAE230A)分析基因表达。在对数据进行标准化和统计分析后,使用Spotfire轮廓搜索和K均值聚类分析,将631个差异表达基因根据其表达水平和动力学分为不同簇。使用实时PCR TaqMan分析对一部分基因基于微阵列的数据进行验证。微阵列数据分析确定了631个差异表达的基因(441个上调和190个下调)(Δ>1.8,P<0.01),显示出特定的基因表达水平和模式。在第-13.8、-13或3天表达增加倍数最高的基因参与趋化作用、炎症反应、细胞黏附和细胞外基质重塑。转录组分析确定了10个上调基因(Δ>5),这些基因以前未与关节炎病理相关联,且位于与自身免疫性疾病相关的基因组区域。大多数下调基因与代谢、转运和肌肉发育调节有关。总之,本研究描述了在Lewis(LEW/N)大鼠SCW诱导的关节炎再激活模型中具有潜在病理生理作用的多个疾病相关基因的时间表达。这些发现增进了我们对该动物模型病理基础分子事件的理解,该模型可能是人类类风湿性关节炎(RA)的一个有价值的对照模型。