Gene expression profiling of IL-1β-stimulated synovial cells from TMJ using Gene Chip
| Project/Area Number |
14571915
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | Nihon University |
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Principal Investigator |
OGURA Naomi Nihon University, School of Dentistry at Matsudo, Lecturer(Full-Time), 松戸歯学部, 講師 (10152448)
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| Co-Investigator(Kenkyū-buntansha) |
HIRRATSUKA Kouichi Nihon University, School of Dentistry at Matsudo, Lecturer(Full-Time), 松戸歯学部, 講師 (80246917)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | temporomandibular joint / synovial fibroblasts / GeneChip / interleukin-1β / Pathway Analysis / 顎関節 / ヒト顎関節滑膜細胞 / GeneChip解析 / Pathway解析 / GeneChip microarray / interluekin-1β / out growth法 / 顎関節症 / ヒト顎間接滑膜細胞 / DNAマイクロアレイ / IL-1β |
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| 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.
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Report
(4 results)
Research Products
(2 results)
