Egr-1 inhibits the expression of extracellular matrix genes in chondrocytes by TNFalpha-induced MEK/ERK signalling.
Egr-1 inhibits the expression of extracellular matrix genes in chondrocytes by TNFalpha-induced MEK/ERK signalling.
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DOI：
10.1186/ar2595
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发表时间：
2009
影响因子：
4.9
通讯作者：
Leask A
中科院分区：
文献类型：
--
作者：
Rockel JS;Bernier SM;Leask A
TNFα在类风湿性关节炎和骨关节炎患者的滑液中升高。TNFα激活软骨细胞中的丝裂原活化激酶激酶（MEK）/细胞外调节激酶（ERK）;然而，软骨细胞中MEK/ERK与TNFα调节的基因表达的总体功能相关性尚不清楚。软骨细胞用TNFα加或不加MEK 1/2抑制剂U 0126处理24小时。微阵列分析和实时PCR分析用于鉴定TNFα以MEK 1/2依赖方式调控的基因。启动子/报告基因、免疫印迹和电泳迁移率变动分析用于鉴定对TNFα应答的活性依赖于MEK 1/2的转录因子。诱饵寡脱氧核苷酸轴承共识转录因子结合位点被引入软骨细胞，以确定我们的结果的功能。软骨细胞中约20%受TNFα调控的基因对U 0126敏感。软骨选择性基质基因Col 2a 1、Agc 1和Hapln 1以及基质金属蛋白酶基因Mmp-12和Mmp-9的转录调控对U 0126敏感，而炎症基因巨噬细胞Csf-1的调控对U 0126不敏感。TNFα诱导的Sox 9和NFκB活性调节也对U 0126不敏感。相反，TNFα增加的早期生长反应1（Egr-1）DNA结合对U 0126敏感。用同源Egr-1寡脱氧核苷酸转染软骨细胞减弱TNFα抑制Col 2a 1、Agc 1或Hapln 1 mRNA表达的能力。我们的研究结果表明，MEK/ERK和Egr 1是TNFα调节的软骨细胞外基质分解代谢和合成代谢基因所必需的，因此可能是骨关节炎或类风湿性关节炎药物干预的潜在靶点。
TNFα is increased in the synovial fluid of patients with rheumatoid arthritis and osteoarthritis. TNFα activates mitogen-activated kinase kinase (MEK)/extracellular regulated kinase (ERK) in chondrocytes; however, the overall functional relevance of MEK/ERK to TNFα-regulated gene expression in chondrocytes is unknown. Chondrocytes were treated with TNFα with or without the MEK1/2 inhibitor U0126 for 24 hours. Microarray analysis and real-time PCR analyses were used to identify genes regulated by TNFα in a MEK1/2-dependent fashion. Promoter/reporter, immunoblot, and electrophoretic mobility shift assays were used to identify transcription factors whose activity in response to TNFα was MEK1/2 dependent. Decoy oligodeoxynucleotides bearing consensus transcription factor binding sites were introduced into chondrocytes to determine the functionality of our results. Approximately 20% of the genes regulated by TNFα in chondrocytes were sensitive to U0126. Transcript regulation of the cartilage-selective matrix genes Col2a1, Agc1 and Hapln1, and of the matrix metalloproteinase genes Mmp-12 and Mmp-9, were U0126 sensitive – whereas regulation of the inflammatory gene macrophage Csf-1 was U0126 insensitive. TNFα-induced regulation of Sox9 and NFκB activity was also U0126 insensitive. Conversely, TNFα-increased early growth response 1 (Egr-1) DNA binding was U0126 sensitive. Transfection of chondrocytes with cognate Egr-1 oligodeoxynucleotides attenuated the ability of TNFα to suppress Col2a1, Agc1 or Hapln1 mRNA expression. Our results suggest that MEK/ERK and Egr1 are required for TNFα-regulated catabolic and anabolic genes of the cartilage extracellular matrix, and hence may represent potential targets for drug intervention in osteoarthritis or rheumatoid arthritis.
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影响因子：
--
作者：
CHU, CQ;FIELD, M;MAINI, RN
通讯作者：
MAINI, RN
影响因子：
4.8
作者：
Kou, I;Ikegawa, S
通讯作者：
Ikegawa, S
影响因子：
4.8
作者：
Favata, MF;Horiuchi, KY;Trzaskos, JM
通讯作者：
Trzaskos, JM
影响因子：
56.9
作者：
FRY, DW;KRAKER, AJ;BRIDGES, AJ
通讯作者：
BRIDGES, AJ
DOI：
10.1016/0925-4439(93)90153-r
发表时间：
1993-08-04
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
CAMPBELL, IK;IANCHES, G;HAMILTON, JA
通讯作者：
HAMILTON, JA