Gene expression profiling of IL-1β-stimulated synovial cells from TMJ using Gene Chip

白细胞介素1受体(IL-1β，IL-1β)在类风湿关节炎等关节疾病的炎症和结缔组织破坏中起重要作用。IL-1β也是关节囊内病理状态的关键介质，包括关节盘移位/内部排列紊乱和骨关节炎。为了从关节炎组织中寻找可能的IL-1β反应基因，我们研究了IL-1β对5例TMJ患者滑膜成纤维细胞8,793个基因表达的影响。方法滑膜成纤维细胞取自1例TMJ关节切开术患者的滑膜组织。用RNasy试剂盒提取滑膜成纤维细胞，加入或不加入I单位/mlIL-1β孵育4h。使用基因芯片(HG Focus Array，8,763个基因)进行基因表达谱分析。杂交数据用Gene Springs？？TM&gt；软件分析。实时定量聚合酶链式反应(Real Time-PCR)检测基因表达水平。结果对照细胞和IL-1β处理后的细胞(P&lt；0.05)共检测到170个基因，其中上调基因139个，下调基因31个。IL-1β增强作用最强的基因是趋化因子CCL20。这170个IL-1β反应基因包括9个趋化因子。实时定量聚合酶链式反应证实，经IL-1β处理后，这些趋化因子的基因表达水平增加。此外，用独创性通径分析方法研究了IL-1β的作用。我们发现IL-1β影响了NF-κB信号通路中多个基因的表达，并通过终点和实时荧光定量分析证实了IL-1β处理后NF-1 B1mRNA和IκBα的表达水平增加。结果提示，IL-1β反应基因在炎症进展和关节成分破坏中起重要作用。

Interleukin-1β(IL-1β) has important roles in the inflammation and connective tissue destruction observed in joint diseases such as rheumatoid arthritis. IL-1β is also a key mediator of intracapsular pathologic conditions of the temporomandibular joint(TMJ), including disk displacement/internal derangement and osteoarthritis. To identify putative IL-1β-responsive genes from arthritic disease tissues, we investigated the effects of IL-1β on gene expression of 8,793 genes in synovial fibroblasts from five TMJ patients. <Methods>Synovial fibroblasts were obtained using an outgrowth method with synovial tissue from a patient who underwent arthrotomy of the TMJ. Total RNA of synovial fibroblasts, incubated with or without i□unit/ml of IL-1β for 4 hours, was extracted using the RNeasy kit. Gene expression pro filing was used GeneChip (HG Focus Array,8,763 genes).Hybridization data were analyzed using Gene Springs^<TM> software. Gene expression levels were also confirmed by real time-PCR. <results>Significant changes between control and IL-1β-treated cells (p<0.05) were detected in 170 genes:139 up-regulated genes and 31 down-regulated genes. The most enhanced gene by IL-1β was CCL20, which is one of chemokines. The 170 IL-1β-responsive genes included 9 chemokines. Real-time PCR confirms that mRNA levels of these chemokines increase after IL-1β treatment. In addition, the effects of IL-1β were investigated using Ingenuity Pathway Analysis. We found that IL-1β affects the expression of several genes in the NFκB signaling pathway and were able to confirm that mRNA levels of NFκB1 and IκBα increase after IL-1β treatment by endpoint-PCR and real time-PCR analysis. The results suggest that IL-1β-responsive genes play important roles in progression of inflammation and destruction of joint components.