The CO-releasing molecule CORM-2 is a novel regulator of the inflammatory process in osteoarthritic chondrocytes
The CO-releasing molecule CORM-2 is a novel regulator of the inflammatory process in osteoarthritic chondrocytes
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DOI：
10.1093/rheumatology/ken264
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发表时间：
2008-09-01
期刊：
影响因子：
5.5
通讯作者：
Alcaraz, M. J.
中科院分区：
文献类型：
--
作者：
Guillen, M. I.;Megias, J.;Alcaraz, M. J.
目标.先前的研究表明，CO释放分子CORM-2可以防止软骨退化。本研究的目的是检测CORM-2是否可以控制骨关节炎软骨细胞中炎症介质的产生并确定相关机制。用IL-1 β刺激来自OA患者的软骨细胞的原代培养物。在存在或不存在CORM-2的情况下测量活性氧、亚硝酸盐、PGE（2）、TNF-α和IL-1受体拮抗剂（IL-1 Ra）的产生。Western blot和实时荧光定量PCR检测细胞中一氧化氮合酶2（NOS 2）、环氧合酶考克斯2（COX 2）和微粒体前列腺素E合酶1（mPGES 1）的表达。ELISA法检测细胞核因子-κ B（NF-κ B）和缺氧诱导因子-1 α（HIF-1 α）的活化及NF-κ B抑制蛋白α（I-κ B α）的磷酸化。CORM-2减少氧化应激、亚硝酸盐和PGE的产生（2）。此外，CORM-2抑制IL-1 β诱导的TNF-α，但增强IL-1 Ra的产生。用CORM-2处理软骨细胞强烈下调NOS-2和mPGES-1蛋白表达，而考克斯-2降低程度较小。这些变化伴随着NOS-2和mPGES-1的mRNA表达的显着下降。CORM-2对p65 NF-κ B B和HIF-1 α的DNA结合活性具有浓度依赖性抑制作用。I κ B α磷酸化也被CORM-2处理降低。这些数据已经为CORM-2打开了新的作用机制，提出了CO释放分子是开发关节疾病新治疗方法的有趣策略的前景。
Objectives. Previous work has shown that the CO-releasing molecule CORM-2 protects against cartilage degradation. The aim of this study was to examine whether CORM-2 can control the production of inflammatory mediators in osteoarthritic chondrocytes and determine the mechanisms involved.Methods. Primary cultures of chondrocytes from OA patients were stimulated with IL-1 beta. The production of reactive oxygen species, nitrite, PGE(2), TNF-alpha and IL-1 receptor antagonist (IL-1Ra) were measured in the presence or absence of CORM-2. The expression of nitric oxide synthase-2 (NOS-2), cyclo-oxygenase-2 (COX-2) and microsomal PG E synthase-1 (mPGES-1) was followed by western blot and real-time PCR. Activation of nuclear factor-kappa B (NF-kappa B) and hypoxia inducible factor-1 alpha (HIF-1 alpha), and phosphorylation of NF-kappa B inhibitory protein alpha (I kappa B alpha) were determined by ELISA.Results. CORM-2 decreased the production of oxidative stress, nitrite and PGE(2). In addition, CORM-2 inhibited IL-1 beta-induced TNF-alpha but enhanced IL-1Ra production. Treatment of chondrocytes with CORM-2 strongly down-regulated NOS-2 and mPGES-1 protein expression, whereas COX-2 was reduced to a lesser extent. These changes were accompanied by a significant decrease in mRNA expression for NOS-2 and mPGES-1. CORM-2 showed a concentration-dependent inhibition of DNA-binding activity for p65 NF-kappa B and HIF-1 alpha. I kappa B alpha phosphorylation was also reduced by CORM-2 treatment.Conclusions. These data have opened new mechanisms of action for CORM-2, raising the prospect that CO-releasing molecules are an interesting strategy for the development of new treatments in articular conditions.