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

2004 Fiscal Year Final Research Report Summary

• Project/Area Number: 14571915
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Surgical dentistry
• Research Institution: Nihon University
• Principal Investigator: OGURA Naomi Nihon University, School of Dentistry at Matsudo, Lecturer(Full-Time), 松戸歯学部, 講師 (10152448)
• Co-Investigator(Kenkyū-buntansha): HIRRATSUKA Kouichi Nihon University, School of Dentistry at Matsudo, Lecturer(Full-Time), 松戸歯学部, 講師 (80246917)
• Project Period (FY): 2002 – 2004
• Keywords: temporomandibular joint / synovial fibroblasts / GeneChip / interleukin-1β / Pathway Analysis

Research Abstract

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.