Protection against cartilage and bone destruction by systemic interleukin-4 treatment in established murine type II collagen-induced arthritis.
Protection against cartilage and bone destruction by systemic interleukin-4 treatment in established murine type II collagen-induced arthritis.
复制标题
在已建立的II型胶原蛋白诱导的关节炎中，全身性白介素4治疗对软骨和骨骼破坏的保护。
DOI：
10.1186/ar14
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
van den Berg , W B
中科院分区：
文献类型：
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作者：
Joosten, L A;Lubberts, E;Helsen, M M;Saxne, T;Coenen-de Roo , C J;Heinegard, D;van den Berg , W B
软骨和骨骼的破坏是人类类风湿关节炎（RA）的标志，控制这些侵蚀过程是类风湿关节炎治疗中最具挑战性的目标。全身白介素-4治疗建立小鼠胶原诱导关节炎抑制疾病活动和保护软骨和骨破坏。血清软骨寡聚基质蛋白（COMP）和组织学检查均证实软骨病理减少。此外，放射学分析显示，骨骼破坏也被部分阻止。当白细胞介素-4治疗与低剂量强的松龙治疗联合使用时，关节肿胀得到了更好的抑制。有趣的是，当低剂量白细胞介素-4与泼尼松龙联合使用时，血清COMP和炎症参数均有协同降低。白细胞介素-4的全身治疗似乎是关节炎软骨和骨骼的一种保护疗法，与低剂量强的松龙联合治疗可能是RA的一种替代疗法。类风湿性关节炎（RA）与一系列细胞因子的产生增加有关，包括肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1，它们显示出强有力的促炎作用，被认为有助于疾病的发病机制。虽然TNF-α似乎是炎症过程中的主要细胞因子，但IL-1是软骨和骨破坏的关键介质。除了直接阻断IL-1/TNF外，还可以在IL-4和IL-10等调节性细胞因子水平上进行调节。IL-4是一种多效性t细胞衍生的细胞因子，可对不同类型的细胞发挥抑制或刺激作用，最初被确定为b细胞生长因子和体液免疫途径的调节剂。IL-4由活化的CD4+ T细胞产生，它促进Th2细胞的成熟。IL-4刺激多种细胞类型的增殖、分化和活化，包括成纤维细胞、内皮细胞和上皮细胞。IL-4也是一种有效的抗炎细胞因子，通过抑制巨噬细胞和单核细胞合成促炎细胞因子，如IL-1、TNF-α、IL-6、IL-8和IL-12。此外，IL-4刺激多种细胞因子抑制剂的合成，如白细胞介素-1受体拮抗剂（IL-1Ra）、可溶性il -1受体II型和TNF受体，IL-4抑制金属蛋白酶的产生，并刺激人单核吞噬细胞和软骨外植体中金属蛋白酶-1组织抑制剂的产生，表明IL-4对细胞外基质降解具有保护作用。此外，IL-4抑制破骨细胞活性和存活，从而阻断体外骨吸收。重要的是，IL-4不能在滑液或组织中检测到。关节中IL-4的缺失可能导致慢性RA中Th1/Th2平衡紊乱。胶原诱导关节炎（CIA）是一种广泛使用的关节炎模型，它显示了人类RA的几个特征。最近有研究表明，CIA的发病受到IL-4和IL-10的严格控制。此外，在免疫阶段暴露于IL-4可降低CIA的发病和严重程度。然而，停止IL-4治疗后，疾病表达增加到控制值。由于有报道称IL-4抑制几种促炎细胞因子和基质降解酶，并上调细胞因子和分解代谢酶的抑制剂，因此我们以已建立的小鼠CIA为模型研究了全身IL-4治疗的组织保护作用。还评估了低剂量抗炎糖皮质激素和IL-4的潜在协同作用。用牛II型胶原免疫DBA-1J/Bom小鼠，并在第21天增强。在免疫后第28天选择已建立CIA的小鼠，用IL-4、强的松龙或强的松龙和IL-4联合治疗数天。目测关节炎评分。采用组织学、影像学和血清软骨寡聚基质蛋白（COMP）评价关节病理。同时测定血清IL-1Ra和抗胶原抗体水平。用IL-4 （1 μg/天）治疗已建立的CIA可抑制疾病活性，如图1所示。令人非常感兴趣的是，尽管1 μg/d IL-4对疾病活动的炎症成分只有中等影响，但组织学检查确定，它强烈地减少了软骨病理（图1）。此外，血清COMP水平显著降低，证实软骨受累减少。此外，组织学和放射学分析均显示骨破坏得以避免（图1）。全身给予IL-4增加血清IL-1Ra水平，降低抗胶原蛋白II型抗体水平。低剂量IL-4 （0.1 μg/d）治疗对疾病评分、血清COMP及关节破坏均无明显抑制作用。当低剂量IL-4与强的松龙（0.05 mg/kg/天）联合使用时，发现了关节炎严重程度和COMP水平的协同抑制，然而，其本身并不有效。在本研究中，我们证明全身IL-4治疗可改善已建立的CIA的疾病进展。虽然临床疾病进展只是被阻止，而不是逆转，但对软骨和骨破坏的明显保护被注意到。这与人类RA和RA动物模型的研究结果一致，表明炎症和组织破坏有时是不耦合的过程。非常重要的是，尽管炎症仍然存在，但暴露于IL-4后发现血清COMP明显降低。这表明血清COMP水平反映软骨损伤，尽管不能排除滑膜炎症的有限贡献。全身用IL-4治疗后发现血清IL-1Ra水平升高（两倍），但这可能不能解释CIA的抑制。我们和其他人已经报道了需要高剂量的IL-1Ra来显著抑制CIA。如前所述，较低剂量的IL-4并没有降低已建立的CIA的临床疾病严重程度。重要的是，低剂量的IL-4和IL-10联合治疗似乎具有更有效的抗炎作用，并显着保护软骨免受破坏。IL-4/强的松龙治疗可改善抗炎效果。此外，还发现了减少软骨和骨破坏的协同效应。这表明全身IL-4/强的松龙治疗可能为人类RA提供软骨和骨骼保护治疗。白细胞介素-4治疗或对照对小鼠疾病活动性、软骨损伤和骨破坏的影响。小鼠分别腹腔注射白介素-4 （IL-4）或1 μg/d。CIA，胶原诱导关节炎。*经Mann-Whitney U检验，P < 0.05。
Destruction of cartilage and bone are hallmarks of human rheumatoid arthritis (RA), and controlling these erosive processes is the most challenging objective in the treatment of RA. Systemic interleukin-4 treatment of established murine collagen-induced arthritis suppressed disease activity and protected against cartilage and bone destruction. Reduced cartilage pathology was confirmed by both decreased serum cartilage oligomeric matrix protein (COMP) and histological examination. In addition, radiological analysis revealed that bone destruction was also partially prevented. Improved suppression of joint swelling was achieved when interleukin-4 treatment was combined with low-dose prednisolone treatment. Interestingly, synergistic reduction of both serum COMP and inflammatory parameters was noted when low-dose interleukin-4 was combined with prednisolone. Systemic treatment with interleukin-4 appeared to be a protective therapy for cartilage and bone in arthritis, and in combination with prednisolone at low dosages may offer an alternative therapy in RA. Rheumatoid arthritis (RA) is associated with an increased production of a range of cytokines including tumour necrosis factor (TNF)-α and interleukin (IL)-1, which display potent proinflammatory actions that are thought to contribute to the pathogenesis of the disease. Although TNF-α seems to be the major cytokine in the inflammatory process, IL-1 is the key mediator with regard to cartilage and bone destruction. Apart from direct blockade of IL-1/TNF, regulation can be exerted at the level of modulatory cytokines such as IL-4 and IL-10. IL-4 is a pleiotropic T-cell derived cytokine that can exert either suppressive or stimulatory effects on different cell types, and was originally identified as a B-cell growth factor and regulator of humoral immune pathways. IL-4 is produced by activated CD4+ T cells and it promotes the maturation of Th2 cells. IL-4 stimulates proliferation, differentiation and activation of several cell types, including fibroblasts, endothelial cells and epithelial cells. IL-4 is also known to be a potent anti-inflammatory cytokine that acts by inhibiting the synthesis of proinflammatory cytokines such as IL-1, TNF-α, IL-6, IL-8 and IL-12 by macrophages and monocytes. Moreover, IL-4 stimulates the synthesis of several cytokine inhibitors such as interleukin-1 receptor antagonist (IL-1Ra), soluble IL-1-receptor type II and TNF receptors IL-4 suppresses metalloproteinase production and stimulates tissue inhibitor of metalloproteinase-1 production in human mononuclear phagocytes and cartilage explants, indicating a protective effect of IL-4 towards extracellular matrix degradation. Furthermore, IL-4 inhibits both osteoclast activity and survival, and thereby blocks bone resorption in vitro. Of great importance is that IL-4 could not be detected in synovial fluid or in tissues. This absence of IL-4 in the joint probably contributes to the disturbance in the Th1/Th2 balance in chronic RA. Collagen-induced arthritis (CIA) is a widely used model of arthritis that displays several features of human RA. Recently it was demonstrated that the onset of CIA is under stringent control of IL-4 and IL-10. Furthermore, it was demonstrated that exposure to IL-4 during the immunization stage reduced onset and severity of CIA. However, after cessation of IL-4 treatment disease expression increased to control values. Because it was reported that IL-4 suppresses several proinflammatory cytokines and matrix degrading enzymes and upregulates inhibitors of both cytokines and catabolic enzymes, we investigated the tissue protective effect of systemic IL-4 treatment using established murine CIA as a model. Potential synergy of low dosages of anti-inflammatory glucocorticosteroids and IL-4 was also evaluated. DBA-1J/Bom mice were immunized with bovine type II collagen and boosted at day 21. Mice with established CIA were selected at day 28 after immunization and treated for days with IL-4, prednisolone, or combinations of prednisolone and IL-4. Arthritis score was monitored visually. Joint pathology was evaluated by histology, radiology and serum cartilage oligomeric matrix protein (COMP). In addition, serum levels of IL-1Ra and anticollagen antibodies were determined. Treatment of established CIA with IL-4 (1 μg/day) resulted in suppression of disease activity as depicted in Figure 1. Of great interest is that, although 1 μg/day IL-4 had only a moderate effect on the inflammatory component of the disease activity, it strongly reduced cartilage pathology, as determined by histological examination (Fig. 1). Moreover, serum COMP levels were significantly reduced, confirming decreased cartilage involvement. In addition, both histological and radiological analysis showed that bone destruction was prevented (Fig. 1). Systemic IL-4 administration increased serum IL-1Ra levels and reduced anticollagen type II antibody levels. Treatment with low-dose IL-4 (0.1 μg/day) was ineffective in suppressing disease score, serum COMP or joint destruction. Synergistic suppression of both arthritis severity and COMP levels was noted when low-dose IL-4 was combined with prednisolone (0.05 mg/kg/day), however, which in itself was not effective. In the present study, we demonstrate that systemic IL-4 treatment ameliorates disease progression of established CIA. Although clinical disease progression was only arrested and not reversed, clear protection against cartilage and bone destruction was noted. This is in accord with findings in both human RA and animal models of RA that show that inflammation and tissue destruction sometimes are uncoupled processes. Of great importance is that, although inflammation was still present, strong reduction in serum COMP was found after exposure to IL-4. This indicated that serum COMP levels reflected cartilage damage, although a limited contribution of the inflamed synovium cannot be excluded. Increased serum IL-1Ra level (twofold) was found after systemic treatment with IL-4, but it is not likely that this could explain the suppression of CIA. We and others have reported that high dosages of IL-1Ra are needed for marked suppression of CIA. As reported previously, lower dosages of IL-4 did not reduce clinical disease severity of established CIA. Of importance is that combined treatment of low dosages of IL-4 and IL-10 appeared to have more potent anti-inflammatory effects, and markedly protected against cartilage destruction. Improved anti-inflammatory effect was achieved with IL-4/prednisolone treatment. In addition, synergistic effects were found for the reduction of cartilage and bone destruction. This indicates that systemic IL-4/prednisolone treatment may provide a cartilage and bone protective therapy for human RA. Effects in mice of treatment with interleukin-4 or control on disease activity, cartilage damage and bone destruction. Mice were treated intraperitoneally for 7 days with either vehicle (control) or 1 μg/day interleukin-4 (IL-4). CIA, collagen-induced arthritis. *P < 0.05, versus control, by Mann-Whitney U test.