Inhibition of interleukin-1β-stimulated production of matrix metalloproteinases by hyaluronan via CD44 in human articular cartilage
Inhibition of interleukin-1β-stimulated production of matrix metalloproteinases by hyaluronan via CD44 in human articular cartilage
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DOI：
10.1002/art.20004
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发表时间：
2004-02-01
影响因子：
--
通讯作者：
Nakamura, T
中科院分区：
文献类型：
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作者：
Julovi, SM;Yasuda, T;Nakamura, T
Objective.探讨透明质酸（HA）抑制白细胞介素1 β（IL 1 β）刺激的人关节软骨基质金属蛋白酶（MMPs）产生的机制。将IL-1 β加入到移植培养的正常和骨关节炎（OA）人关节软骨中以刺激MMP的产生。将关节软骨与临床使用的800-kd HA一起孵育或预孵育，以评估其对IL-1 β诱导的MMPs的作用。通过免疫印迹法检测条件培养基中分泌的MMPs 1，3和13的水平;通过免疫荧光显微镜评价软骨细胞中的细胞内MMP合成。使用荧光素化的HA通过荧光显微镜分析HA向软骨组织中的渗透及其与CD 44的结合。用抗CD 44抗体进行阻断实验，探讨HA的作用机制。在正常和OA软骨外植体培养中，用1 mg/ml的800 kd HA处理和预处理导致IL-1 β刺激的MMPs 1、3和13的产生显著抑制。荧光组织化学显示HA穿透软骨组织并定位于软骨细胞周围的细胞周基质中。使用抗CD 44抗体的HA结合阻断实验证明HA与软骨细胞的缔合是由CD 44介导的。抗CD 44抗体可抑制IL-1 β刺激的MMP的产生，而RA对IL-1 β诱导的正常和OA软骨MMP产生的抑制作用可逆转。这项研究表明，HA有效地抑制IL-1 β刺激的MMP-1，MMP-3和MMP-13的产生，这支持了RA在OA治疗中的临床应用。HA对IL-1 β的作用可能涉及RA与软骨细胞上的CD 44之间的直接相互作用。
Objective. To investigate the mechanism of the inhibitory action of hyaluronan (HA) on interleukin-1beta (IL-1beta)-stimulated production of matrix metalloproteinases (MMPs) in human articular cartilage.Methods. IL-1beta was added to normal and osteoarthritic (OA) human articular cartilage in explant culture to stimulate MMP production. Articular cartilage was incubated or preincubated with a clinically used form of 800-kd HA to assess its effect on IL-1beta-induced MMPs. Levels of secreted MMPs 1, 3, and 13 in conditioned media were detected by immunoblotting; intracellular MMP synthesis in chondrocytes was evaluated by immunofluorescence microscopy. Penetration of HA into cartilage tissue and its binding to CD44 were analyzed by fluorescence microscopy using fluoresceinated HA. Blocking experiments with anti-CD44 antibody were performed to investigate the mechanism of action of HA.Results. Treatment and pretreatment with 800-kd HA at 1 mg/ml resulted in significant suppression of IL-1beta-stimulated production of MMPs 1, 3, and 13 in normal and OA cartilage explant culture. Fluorescence histocytochemistry revealed that HA penetrated cartilage tissue and localized in the pericellular matrix around chondrocytes. HA-binding blocking experiments using anti-CD44 antibody demonstrated that the association of HA with chondrocytes was mediated by CD44. Preincubation with anti-CD44 antibody, which suppressed IL-1beta-stimulated MMPs, reversed the inhibitory effect of RA on MMP production that was induced by IL-1beta in normal and OA cartilage.Conclusion. This study demonstrates that HA effectively inhibits IL-1beta-stimulated production of MMP-1, MMP-3, and MMP-13, which supports the clinical use of RA in the treatment of OA. The action of HA on IL-1beta may involve direct interaction between RA and CD44 on chondrocytes.