Attachment to laminin-111 facilitates transforming growth factor β-induced expression of matrix metalloproteinase-3 in synovial fibroblasts
Attachment to laminin-111 facilitates transforming growth factor β-induced expression of matrix metalloproteinase-3 in synovial fibroblasts
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DOI：
10.1136/ard.2006.060228
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发表时间：
2007-04-01
影响因子：
27.4
通讯作者：
Aicher, Wilhelm K.
中科院分区：
文献类型：
--
作者：
Hoberg, Maik;Rudert, Maximilian;Aicher, Wilhelm K.
背景资料：在类风湿性关节炎（RA）患者的滑膜中，层粘连蛋白和基质降解蛋白酶的强表达被报道。目的：研究层粘连蛋白-1（LM- 111）对骨关节炎（OA）和类风湿关节炎（RA）患者滑膜成纤维细胞（SF）基质金属蛋白酶（MMPs）表达的调节作用，以及TGF-β共刺激信号的存在与否。采用定量逆转录-聚合酶链反应（RT-PCR）检测基因表达免疫印迹法检测细胞内信号转导通路，SB 203580、PD 98059和A- 83- 01分别对p38 MAPK、MEK-ERK和SMAD 2进行毒性作用。SF与LM- 111的连接没有激活MMP的表达，但加入TGF β诱导MMP- 3的表达增加了5倍。在存在TGF β的情况下，SF在LM- 111上的孵育诱导MMP- 3 mRNA的显著12倍高表达，并且MMP- 3的分泌比对照升高20倍。功能性阻断LM- 111 -整合素相互作用可显著降低层粘连蛋白激活的MMP- 3表达。LM- 111和TGFb刺激SF可激活p38 MAPK、ERK和SMAD 2通路，SB 203580、PD 98059和A- 83- 01抑制这些通路证实这些通路参与MMP-3的调节。SF与LM- 111的连接本身对MMP- 1或MMP-3的表达只有很小的影响，但它显著促进TGF β诱导的MMP- 3的表达。因此，这种MMP- 3诱导模式可能导致RA中的炎性关节破坏，而不依赖于促炎细胞因子白细胞介素（IL）1 β或肿瘤坏死因子（TNF）α。
Background: In the synovial membrane of patients with rheumatoid arthritis ( RA), a strong expression of laminins and matrix degrading proteases was reported. Aim: To investigate the regulation of matrix metalloproteinases ( MMPs) in synovial fibroblasts ( SFs) of patients with osteoarthritis ( OA) and RA by attachment to laminin- 1 ( LM- 111) and in the presence or absence of costimulatory signals provided by transforming growth factor beta ( TGF beta).Methods: SFs were seeded in laminin- coated flasks and activated by addition of TGF beta. The expression of genes was investigated by quantitative reverse transcriptase- polymerase chain reaction ( qRT- PCR), immunocytochemistry and ELISA, and intracellular signalling pathways by immunoblotting, and by poisoning p38MAPK by SB203580, MEK- ERK by PD98059 and SMAD2 by A- 83- 01.Results: Attachment of SF to LM- 111 did not activate the expression of MMPs, but addition of TGFb induced a fivefold higher expression of MMP- 3. Incubation of SF on LM- 111 in the presence of TGFb induced a significant 12- fold higher expression of MMP- 3 mRNA, and secretion of MMP- 3 was elevated 20- fold above controls. Functional blocking of LM- 111 - integrin interaction reduced the laminin- activated MMP- 3 expression significantly. Stimulation of SF by LM- 111 and TGFb activated the p38MAPK, ERK and SMAD2 pathways, and inhibition of these pathways by using SB203580, PD98059 or A- 83- 01 confirmed the involvement of these pathways in the regulation of MMP- 3.Conclusion: Attachment of SF to LM- 111 by itself has only minor effects on the expression of MMP- 1 or MMP3, but it facilitates the TGF beta-induced expression of MMP- 3 significantly. This mode of MMP- 3 induction may therefore contribute to inflammatory joint destruction in RA independent of the proinflammatory cytokines interleukin ( IL)1 beta or tumour necrosis factor ( TNF)alpha.