Stress-induced signaling pathways in hyalin chondrocytes: inhibition by Avocado-Soybean Unsaponifiables (ASU)
Stress-induced signaling pathways in hyalin chondrocytes: inhibition by Avocado-Soybean Unsaponifiables (ASU)
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DOI：
10.1016/j.joca.2007.06.016
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发表时间：
2008-03-01
影响因子：
7
通讯作者：
Berenbaum, F.
中科院分区：
文献类型：
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作者：
Gabay, O.;Gosset, M.;Berenbaum, F.
目的：鳄梨大豆不皂化物（ASU）是治疗症状性骨关节炎（OA）最常用的药物之一。其活动机制仍知之甚少。我们在这里研究 ASU 对小鼠或人类软骨细胞信号通路的影响。方法：在存在或不存在 ASU (10 μg/ml) 的情况下，研究了用白细胞介素 1 β (IL1 β，10 ng/ml) 刺激的小鼠或人类软骨细胞以及受到压缩机械应力 (MS) 的软骨。使用 1-kappa B α 抗体通过免疫印迹评估核因子 kappa B (NF-kappa B) 激活，使用 p65 抗体评估 NF-kappa B 核转位，并使用磷酸和 ERK1/2 抗体评估细胞外信号调节激酶 (ERK)1/2 激活。通过电泳迁移率变动分析研究了 p50/p65 复合物与 DNA 的结合。结果：在我们的模型中，ASU 减少了基质金属蛋白酶-3 和-13 的表达以及前列腺素 E-2 (PGE(2)) 的释放。当软骨细胞受到 IL1 β 或 MS 刺激时，1-kappa B α 蛋白在细胞质中持续表达，表明 ASU 存在时可阻止 1-kappa B α 的降解。 NF-κ B 复合物的核转位表现为胞浆中 p65 蛋白的减少，而 p65 在 IL1 β 刺激下出现在细胞核中。当亚利桑那州立大学存在时，这种易位就被废除了。此外，带移实验表明，ASU 会抑制 IL1 β 诱导的 p50/p65 复合物与 NF-κ B 反应元件的结合。最后，在已知由 IL1 β 诱导的不同丝裂原激活蛋白激酶中，ERK1/2 是唯一被 ASU 抑制的激酶。结论：这些结果表明 ASU 表达一系列独特的活性，可以抵消 OA 中涉及的有害过程，例如炎症。 (C) 2007 年国际骨关节炎研究协会。由爱思唯尔有限公司出版。保留所有权利。
Objective: Avocado-Soybean Unsaponifiables (ASU) represent one of the most commonly used drugs for symptomatic osteoarthritis (OA). The mechanisms of its activities are still poorly understood. We investigate here the effects of ASU on signaling pathways in mouse or human chondrocytes.Methods: Mouse or human chondrocytes stimulated with interleukin-1 beta (IL1 beta, 10 ng/ml) and cartilage submitted to a compressive mechanical stress (MS) were studied in the presence or absence of ASU (10 mu g/ml). Nuclear factor kappa B (NF-kappa B) activation was assessed by immunoblot, using an 1-kappa B alpha antibody, nuclear translocation of NF-kappa B using p65 antibody, and extra-cellular signal-regulated kinase (ERK)1/2 activation using phospho and ERK1/2 antibodies. The binding of the p50/p65 complex on DNA was studied by electrophoretic mobility shift assay.Results: ASU decrease matrix metalloproteinases-3 and -13 expressions and Prostaglandin E-2 (PGE(2)) release in our model. The degradation of 1-kappa B alpha is prevented in the presence of ASU as shown by the persistent expression of 1-kappa B alpha protein in the cytosol when chondrocytes are stimulated by IL1 beta or MS. Nuclear translocation of the NF-kappa B complex is shown by the decrease of the p65 protein from the cytosol, whereas p65 appears in the nucleus under IL1 beta stimulation. This translocation is abolished in the presence of ASU. Moreover, bandshift experiments show an inhibition of the IL1 beta-induced binding of p50/p65 complexes to NF-kappa B responsive elements in response to ASU. Finally, among the different mitogen-activated protein kinases known to be induced by IL1 beta, ERK1/2 was the sole kinase inhibited by ASU.Conclusion: These results demonstrate that ASU express a unique range of activities, which could counteract deleterious processes involved in OA, such as inflammation. (C) 2007 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.