Interleukin (IL)-23 p19 expression induced by IL-1β in human fibroblast-like synoviocytes with rheumatoid arthritis via active nuclear factor-κB and AP-1 dependent pathway
Interleukin (IL)-23 p19 expression induced by IL-1β in human fibroblast-like synoviocytes with rheumatoid arthritis via active nuclear factor-κB and AP-1 dependent pathway
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DOI：
10.1093/rheumatology/kem055
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发表时间：
2007-08-01
期刊：
影响因子：
5.5
通讯作者：
Chang, D.-M.
中科院分区：
文献类型：
--
作者：
Liu, F.-L.;Chen, C.-H.;Chang, D.-M.
目标.目的探讨类风湿关节炎（RA）关节中白细胞介素-23（IL-23）p19亚单位的来源，IL-1 β和肿瘤坏死因子（TNF）-α对RA成纤维细胞样滑膜细胞IL-23基因表达的影响，以及IL-23对促炎细胞因子的影响。采用免疫组织化学方法检测RA和骨关节炎（OA）患者关节中IL-23 p19的表达。采用逆转录聚合酶链反应（RT-PCR）、定量PCR和Western blotting检测IL-1 β和TNF-α对类风湿关节炎（RA）患者成纤维样滑膜细胞（HFLS RA）IL-23 p19和IL-12 p35亚单位表达的影响。核因子κ B（NF-κ B）或AP-1激活的阻断用于验证p19诱导的细胞内信号通路的参与。采用RT-PCR和酶联免疫吸附法检测IL-23诱导HFLS RA产生IL-8和IL-6。IL-23 p19在RA患者滑膜中表达，而在OA患者滑膜中不表达。与蛋白表达相似，5例RA患者关节液单个核细胞（SFMC）中有4例可检测到IL-23 p19 mRNA。IL-1 β和TNF-α可诱导RA成纤维细胞样滑膜细胞产生IL-23 p19亚基。IL-1 β的作用比TNF-α强得多。观察到这些反应具有剂量反应性和时间依赖性。IL-1 β产生IL-12的p35亚基的微弱增强的基因表达。IL-1 β，8也促进p35的表达，p35是IL-12的亚基，但较弱。此外，NF-κ B和AP-1抑制剂下调IL-1 β诱导的IL-23 p19 mRNA的表达。IL-23受体（IL-23 R）在HFLS RA中呈组成性表达。IL-23可剂量依赖性上调HFLS RA中IL-8和IL-6的mRNA和蛋白水平。我们的结果表明，RA和HFLS-RA滑液中的单核细胞产生的IL-23通过诱导HFLS产生IL-8和IL-6来促进RA的炎症反应。IL-1 β，8通过NF-κ B和AP-1途径调节IL-23 p19表达。该报告还表明，IL-23可以通过刺激IL-8和IL-6的产生来促进HFLS-RA的炎症反应。
Objectives. To explore the source of the p19 subunit of interleukin-23 (IL-23) in joints with rheumatoid arthritis (RA), the effects of IL-1 beta and tumour necrosis factor (TNF)-alpha on IL-23 gene expression in RA fibroblast-like synoviocytes and the effect of IL-23 on proinflammatory cytokines.Methods. Expression of IL-23 p19 in joints was examined by immunohistochemical analysis of patients with RA and osteoarthritis (OA). The effects of IL-1 beta and TNF-alpha on the expression, of IL-23 p19 and IL-12 p35 subunits in human fibroblast-like synoviocytes from RA patients (HFLS-RA) were determined by reverse transcriptase polymerase chain reaction (RT-PCR), quantitative PCR and western blotting assay. Blockade of nuclear factor kappaB (NF-kappa B) or AP-1 activation was used to verify the involvement of intracellular signal pathways of the induction of p19. IL-23-induced IL-8 and IL-6 productions were determined in HFLS-RA by RT-PCR and enzyme-linked immunosorbent assay.Results. IL-23 p19 was expressed in the synovium from RA, but not from OA patients. Similar to the protein expression, IL-23 p19 mRNA could be detected by RT-PCR in four of five RA synovial fluid mononuclear cells (SFMC). IL-1 beta and TNF-alpha could induce RA fibroblast-like synoviocytes to produce the IL-23 p19 subunit. The effects of IL-1 beta ere much stronger than TNF-alpha. These responses were observed in both a dose-responsive and time-dependent manner. IL-1 beta produced weakly enhanced gene expression of the p35 subunits of IL-12. IL-1 beta,8 also promotes the p35 expression, a subunit of IL-12, but weakly. In addition, the NF-kappa B and the AP-1 inhibitors down-regulated the expression of IL-23 p19 mRNA induced by IL-1 beta . IL-23 receptor (IL-23R) was of constitutive expression in HFLS-RA. Moreover, IL-23 up-regulated the IL-8 and IL-6 mRNA and protein levels in a dose-dependent manner in HFLS-RA.Conclusions. Our results demonstrate that IL-23, produced by mononuclear cells in synovial fluid with RA and HFLS-RA, promotes inflammatory responses in RA by inducing IL-8 and IL-6 production from HFLS. IL-1 beta,8 regulates IL-23 p19 expression via NF-kappa B and AP-1 pathways. This report also demonstrates that IL-23 could promote inflammatory responses in HFLS-RA by stimulating IL-8 and IL-6 production.