Sandell L J, Xing X, Franz C, Davies S, Chang L-W, Patra D
Department of Orthopaedic Surgery, Washington University School of Medicine at Barnes-Jewish Hospital, St Louis, MO 63110, United States.
Osteoarthritis Cartilage. 2008 Dec;16(12):1560-71. doi: 10.1016/j.joca.2008.04.027. Epub 2008 Jun 18.
To provide a more complete picture of the effect of interleukin-1 beta (IL-1beta) on adult human articular chondrocyte gene expression, in contrast to the candidate gene approach.
Chondrocytes from human knee cartilage were cultured in medium containing IL-1beta. Changes in gene expression were analyzed by microarray and reverse transcriptase-polymerase chain reaction analysis. The ability of transforming growth factor beta-1 (TGF-beta1), fibroblast growth factor (FGF)-18, and bone morphogenetic protein 2 (BMP-2) to alter the effects of IL-1beta was analyzed. Computational analysis of the promoter regions of differentially expressed genes for transcription factor binding motifs was performed.
IL-1beta-treated human chondrocytes showed significant increases in the expression of granulocyte colony stimulating factor-3, endothelial leukocyte adhesion molecule 1 and leukemia inhibitory factor as well as for a large group of chemokines that include CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL4, CCL5, CCL8, CCL20, CCL3L1, CX3CL1 and the cytokine IL-6. As expected, the mRNA for matrix metalloproteinase (MMP)-13 and BMP-2 also increased while mRNA for the matrix genes COL2A1 and aggrecan was down-regulated. A subset of chemokines increased rapidly at very low levels of IL-1beta. The phenotype induced by IL-1beta was partially reversed by TGF-beta1, but not by BMP-2. In the presence of IL-1beta, FGF-18 increased expression of ADAMTS-4, aggrecan, BMP-2, COL2A1, CCL3, CCL4, CCL20, CXCL1, CXCL3, CXCL6, IL-1beta, IL-6, and IL-8 and decreased ADAMTS-5, MMP-13, CCL2, and CCL8. Computational analysis revealed a high likelihood that the most up-regulated chemokines are regulated by the transcription factors myocyte enhancer binding factor-3 (MEF-3), CCAAT/enhancer binding protein (C/EBP) and nuclear factor-kappa B (NF-kappaB).
IL-1beta has a diverse effect on gene expression profile in human chondrocytes affecting matrix genes as well as chemokines and cytokines. TGF-beta1 has the ability to antagonize some of the phenotype induced by IL-1beta.
与候选基因方法相比，更全面地描绘白细胞介素 -1β（IL-1β）对成人人类关节软骨细胞基因表达的影响。
将来自人膝关节软骨的软骨细胞在含有IL-1β的培养基中培养。通过微阵列和逆转录聚合酶链反应分析来分析基因表达的变化。分析转化生长因子β-1（TGF-β1）、成纤维细胞生长因子（FGF）-18和骨形态发生蛋白2（BMP-2）改变IL-1β作用的能力。对差异表达基因的启动子区域进行转录因子结合基序的计算分析。
经IL-1β处理的人软骨细胞中，粒细胞集落刺激因子-3、内皮白细胞黏附分子1和白血病抑制因子以及一大组趋化因子（包括CXCL1、CXCL2、CXCL3、CXCL5、CXCL6、CXCL8、CCL2、CCL3、CCL4、CCL5、CCL8、CCL20、CCL3L1、CX3CL1和细胞因子IL-6）的表达显著增加。正如预期的那样，基质金属蛋白酶（MMP）-13和BMP-2的mRNA也增加，而基质基因COL2A1和聚集蛋白聚糖的mRNA则下调。一部分趋化因子在极低水平的IL-1β作用下迅速增加。IL-1β诱导的表型被TGF-β1部分逆转，但未被BMP-2逆转。在存在IL-1β的情况下，FGF-18增加了ADAMTS-4、聚集蛋白聚糖、BMP-2、COL2A1、CCL3、CCL4、CCL20、CXCL1、CXCL3、CXCL6、IL-1β、IL-6和IL-8的表达，并降低了ADAMTS-5、MMP-13、CCL2和CCL8的表达。计算分析表明，上调最明显的趋化因子极有可能受转录因子肌细胞增强子结合因子-3（MEF-3）、CCAAT/增强子结合蛋白（C/EBP）和核因子-κB（NF-κB）调控。
IL-1β对人软骨细胞的基因表达谱具有多种影响，既影响基质基因，也影响趋化因子和细胞因子。TGF-β1具有拮抗IL-1β诱导的部分表型的能力。
