Lauder S N, Carty S M, Carpenter C E, Hill R J, Talamas F, Bondeson J, Brennan P, Williams A S
Rheumatology Research Laboratory, Cardiff University, Heath Park, Cardiff CF14 4XN, UK.
Rheumatology (Oxford). 2007 May;46(5):752-8. doi: 10.1093/rheumatology/kel419. Epub 2007 Jan 11.
To investigate the importance of activation of the transcription factor, nuclear factor-kappaB (NF-kappaB) by interleukin-1beta (IL-1beta) and tumour necrosis factor-alpha (TNF-alpha) in the pathogenesis of osteoarthritis (OA) and assess its suitability as a target for therapy by determining its role in the induction of the cytokine IL-6 and the degenerative enzymes, matrix metalloproteinase (MMP)-1 and MMP-3 in vitro.
Three distinct cellular models, derived from primary OA tissue, were employed, namely, fibroblast-like synoviocytes (OA-SF); co-cultures containing phenotypic macrophage-like and fibroblast-like cells (OA-COCUL); and primary OA synovial tissue explants (OA-EXP). These were treated with specific inhibitors of IL-1beta, TNF-alpha and NF-kappaB to assess their differential role in the production of pathologically relevant mediators, specifically IL-6, MMP-1, MMP-3 and the tissue inhibitor of metalloproteinases-1 (TIMP-1), which were quantified by enzyme-linked immunosorbent assay.
Inhibition of NF-kappaB by a novel agent, RO100 at a dose of 0.1 microM, exerted significant (P < 0.05) repression of IL-6, MMP-1 and MMP-3 production in OA-SF. Notably, neither TIMP-1 production nor cell viability was significantly affected at the dose tested. These data were reproduced in OA-EXP, which might be considered as having greater physiological relevance. Interestingly, comparable efficacy was noted using IL-1beta and TNF-alpha neutralizing antibodies in OA-COCUL.
We have demonstrated that a novel pharmacological inhibitor of NF-kappaB, RO100 inhibits pathological mediators of OA progression with equivalent efficacy as established IL-1beta and TNF-alpha neutralizing strategies. Our findings highlight a potential for developing NF-kappaB targeted therapeutics for positively regulating disease activity and improving clinical outcome in OA.
研究白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）激活转录因子核因子-κB（NF-κB）在骨关节炎（OA）发病机制中的重要性，并通过确定其在体外诱导细胞因子IL-6和退行性酶基质金属蛋白酶（MMP）-1及MMP-3中的作用，评估其作为治疗靶点的适用性。
采用三种源自原发性OA组织的不同细胞模型，即成纤维细胞样滑膜细胞（OA-SF）；含有表型巨噬细胞样和成纤维细胞样细胞的共培养物（OA-COCUL）；以及原发性OA滑膜组织外植体（OA-EXP）。用IL-1β、TNF-α和NF-κB的特异性抑制剂处理这些模型，以评估它们在产生病理相关介质（特别是IL-6、MMP-1、MMP-3和金属蛋白酶组织抑制剂-1（TIMP-1））中的不同作用，这些介质通过酶联免疫吸附测定法定量。
新型药物RO100以0.1微摩尔/升的剂量抑制NF-κB，对OA-SF中IL-6、MMP-1和MMP-3的产生有显著（P<0.05）抑制作用。值得注意的是，在所测试的剂量下，TIMP-1的产生和细胞活力均未受到显著影响。这些数据在OA-EXP中得到重现，OA-EXP可能被认为具有更大的生理相关性。有趣的是，在OA-COCUL中使用IL-1β和TNF-α中和抗体也观察到了类似的效果。
我们已经证明，新型NF-κB药理学抑制剂RO100抑制OA进展的病理介质，其效果与既定的IL-1β和TNF-α中和策略相当。我们的研究结果突出了开发针对NF-κB的疗法以正向调节疾病活动并改善OA临床结局的潜力。
