CCL2 induces articular chondrocyte MMP expression through ERK and p38 signaling pathways.
CCL2 induces articular chondrocyte MMP expression through ERK and p38 signaling pathways.
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CCL2通过ERK和p38信号通路诱导关节软骨细胞MMP表达。
DOI：
10.1016/j.ocarto.2020.100136
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发表时间：
2021-03
期刊：
影响因子：
--
通讯作者：
Longobardi, Lara
中科院分区：
文献类型：
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作者：
Willcockson, Helen;Ozkan, Huseyin;Chubinskaya, Susan;Loeser, Richard F;Longobardi, Lara
在之前的研究中，我们确定了血清和关节软骨CCL2水平升高与骨关节炎（OA）进展、软骨损伤和软骨中MMP13升高之间的关联。在这里，我们分析了健康和OA人关节软骨细胞中导致软骨分解代谢标志物基因表达的CCL2下游信号介质。用重组人CCL2或不用重组人CCL2处理健康人或OA受试者的人关节软骨细胞；收集细胞裂解液或mRNA进行免疫印迹或qRT-PCR。为了进行途径分析，在CCL2治疗之前，将软骨细胞与CCR2（独特的CCL2受体）抑制剂、ERK抑制剂或p38抑制剂一起预孵育。CCR2治疗健康和OA软骨细胞通过CCR2激活ERK和p38。在健康软骨细胞中，短时间（6h）和长时间（24-72h） CCL2治疗导致Ccr2、Mmp-1、Mmp-3、Mmp-13和Timp1上调。在OA软骨细胞中，CCL2诱导Ccr2、Mmp-1和Mmp-3的表达，但不诱导Mmp1和Timp1的表达，且仅在较长时间（72h）后表达。在健康和OA软骨细胞中，Ccr2介导的Ccr2和软骨分解代谢标志物的上调是由ERK和p38信号介导的。关节软骨细胞中CCL2/CCR2轴的触发激活特定的MAPK通路，导致软骨降解酶的基因表达。然而，健康软骨细胞与OA软骨细胞对CCL2刺激的反应在激活基因数量和暴露于CCL2的时间方面存在一些差异，这表明CCL2在关节软骨中的作用可能取决于OA的分期和严重程度。
In previous studies, we determined an association between increased serum and articular cartilage levels of CCL2 with osteoarthritis (OA) progression, cartilage damage and increased MMP13 in cartilage. Here we analyzed CCL2 downstream signaling mediators that lead to gene expression of cartilage catabolic markers, in healthy and OA human articular chondrocytes. Human articular chondrocytes obtained from healthy or OA subjects were treated with or without recombinant human CCL2; cell lysates or mRNA were collected for immunoblotting or qRT-PCR. For pathway analysis, chondrocytes were pre-incubated with an inhibitor of CCR2 (the unique CCL2 receptor), ERK inhibitor or p38 inhibitor prior to CCL2 treatment. CCL2 treatment of both healthy and OA chondrocytes activated ERK and p38 via CCR2. In healthy chondrocytes, short (6h) and prolonged (24–72h) CCL2 treatments led to Ccr2, Mmp-1, Mmp-3, Mmp-13 and Timp1 upregulation. In OA chondrocytes, CCL2 induced expression of Ccr2, Mmp-1 and Mmp-3, but not Mmp1 and Timp1, and only following longer treatments (72h). In both healthy and OA chondrocytes, the CCL2-mediated upregulation of Ccr2 and cartilage catabolic markers was mediated by ERK and p38 signaling. The triggering of the CCL2/CCR2 axis in articular chondrocytes activates specific MAPK pathways leading to gene expression of cartilage degrading enzymes. However, some differences in the response to CCL2 stimulation are detected in healthy vs OA chondrocytes with respect to the number of activated genes and to the time of exposure to CCL2, suggesting that CCL2 action in articular cartilage may be dependent on OA stage and severity.