Lectin-like oxidized low-density lipoprotein receptor 1 mediates matrix metalloproteinase 3 synthesis enhanced by oxidized low-density lipoprotein in rheumatoid arthritis cartilage
Lectin-like oxidized low-density lipoprotein receptor 1 mediates matrix metalloproteinase 3 synthesis enhanced by oxidized low-density lipoprotein in rheumatoid arthritis cartilage
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DOI：
10.1002/art.20581
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发表时间：
2004-11-01
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：
Kakinuma, T;Yasuda, T;Nakamura, T
Objective.研究氧化低密度脂蛋白（ox-LDL）和凝集素样氧化低密度脂蛋白受体1（LOX-1）在类风湿关节炎（RA）关节软骨标本中的存在，并确定ox-LDL与LOX-1的相互作用是否能诱导关节软骨组织培养中基质金属蛋白酶3（MMP-3）的表达。从RA、骨关节炎（OA）和股骨颈骨折患者获得的人关节软骨标本通过共聚焦荧光显微镜检查LOX-1和ox-LDL。通过免疫荧光分析来评估ox-LDL和LOX-1之间的关联。股骨颈骨折患者的关节软骨标本与ox-LDL孵育，有或没有与中和抗LOX-1抗体预孵育。采用免疫荧光法检测软骨细胞MMP-3的合成情况，采用免疫印迹法和酶联免疫吸附法检测MMP-3向条件培养液中的分泌情况。大多数RA软骨细胞与抗LOX-1和抗ox-LDL抗体均呈阳性染色;然而，在OA和正常软骨标本中未发现阳性细胞。抗LOX-1抗体可抑制Dil标记的ox-LDL与软骨细胞的结合，表明这种相互作用是由LOX-1介导的。与天然LDL相比，ox-LDL诱导关节软骨细胞合成MMP-3，并诱导LOX-1，从而导致MMP-3分泌到培养基中。抗LOX-1抗体逆转ox-LDL刺激的MMP-3合成至对照水平。结论Ox-LDL主要由LOX-1介导，增强关节软骨细胞MMP-3的产生。RA软骨中ox-LDL的积累增加，LOX-1的表达升高，表明RA软骨病理中受体-配体相互作用的特定作用。
Objective. To investigate for the presence of oxidized low-density lipoprotein (ox-LDL) and lectin-like oxidized LDL receptor 1 (LOX-1) in cartilage specimens from rheumatoid arthritis (RA) joints and to determine whether the interaction of ox-LDL with LOX-1 can induce matrix metalloproteinase 3 (MMP-3) in articular cartilage explant culture.Methods. Human articular cartilage specimens obtained from patients with RA, osteoarthritis (OA), and femoral neck fractures were examined for LOX-1 and ox-LDL by confocal fluorescence microscopy. The association between ox-LDL and LOX-1 was evaluated by immunofluorescence analysis. Articular cartilage specimens from patients with femoral neck fractures were incubated with ox-LDL, with or without preincubation with neutralizing anti-LOX-1 antibody. MMP-3 synthesis by chondrocytes in explant cartilage was evaluated by immunofluorescence, and protein secretion into conditioned medium was monitored by immunoblotting and enzyme-linked immunosorbent assay.Results. The majority of the RA chondrocytes stained positively with both anti-LOX-1 and anti-ox-LDL antibodies; however, no positive cells were found in OA and normal cartilage specimens. Anti-LOX-1 antibody suppressed the binding of Dil-labeled ox-LDL to chondrocytes in explant culture, suggesting that the interaction was mediated by LOX-1. In contrast to native LDL, ox-LDL induced MMP-3 synthesis by articular chondrocytes in association with the induction of LOX-1, which resulted in enhanced secretion of MMP-3 into the culture medium. Anti-LOX-1 antibody reversed ox-LDL-stimulated MMP-3 synthesis to control levels. Conclusion. Ox-LDL, principally mediated by LOX-1, enhanced MMP-3 production in articular chondrocytes. Increased accumulation of ox-LDL with elevated expression of, LOX-1 in RA cartilage indicates a specific role of the receptor-ligand interaction in cartilage pathology in RA.