CC and CXC chemokine receptors mediate migration, proliferation, and matrix metalloproteinase production by fibroblast-like synoviocytes from rheumatoid arthritis patients
CC and CXC chemokine receptors mediate migration, proliferation, and matrix metalloproteinase production by fibroblast-like synoviocytes from rheumatoid arthritis patients
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DOI：
10.1002/art.20615
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发表时间：
2004-12-01
影响因子：
--
通讯作者：
Díaz-González, F
中科院分区：
文献类型：
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作者：
García-Vicuña, R;Gómez-Gaviro, MV;Díaz-González, F
客观的。为了探索趋化因子系统在类风湿关节炎（RA）滑膜细胞介导的组织破坏中的潜在参与，我们研究了趋化因子受体的表达谱及其在来自RA患者的培养的成纤维样滑膜细胞（FLS）的迁移、增殖和基质金属蛋白酶（MMP）产生中的功能。方法。通过逆转录酶-聚合酶链反应在信使 RNA (mRNA) 水平上研究培养的 FLS 上 CC 和 CXC 趋化因子受体的存在，并通过流式细胞术在细胞表面表达水平上研究。通过 Fura Red 预载 FLS 上的流式细胞术评估趋化因子刺激诱导的细胞质钙流入的变化。两室 Transwell 室用于 FLS 趋化性测定。通过基于荧光的增殖测定来测量细胞生长。通过原纤维降解测定和酶谱法测定明胶酶和胶原酶活性。结果。 FLS在细胞表面和mRNA水平上组成型表达受体CCR2、CCR5、CXCR3和CXCR4，但未能表达CCR3和CCR6。在用单核细胞趋化蛋白 1 (MCP-1)、基质细胞衍生因子 1a (SDF-1α) 和干扰素诱导蛋白 10 (IP-10) 攻击的 FLS 上观察到显着的胞质内钙流入。 MCP-1、SDF-1α、IP-10 和干扰素-γ 诱导的单核因子的刺激增强了 FLS 的迁移和增殖。除了 RANTES 之外，这些趋化因子还以剂量和时间依赖性方式增加培养的 FLS 无细胞上清液中的明胶酶和胶原酶活性。有趣的是，趋化因子介导的 MMP 活性上调被抗白细胞介素 1β 阻断抗体的存在显着消除，但抗肿瘤坏死因子 α 阻断抗体却没有显着消除。结论。这些数据表明，通过 FLS 调节迁移、增殖和 MNIP 产生，趋化因子系统可能在 RA 的破坏阶段发挥比目前怀疑的更直接的作用，从而强调趋化因子及其受体作为该疾病潜在治疗靶点的相关性。
Objective. To explore the potential involvement of the chemokine system in synoviocyte-mediated tissue destruction in rheumatoid arthritis (RA), we studied the expression profile of chemokine receptors and their function in the migration, proliferation, and matrix metalloproteinase (MMP) production of cultured fibroblast-like synoviocytes (FLS) from RA patients.Methods. The presence of CC and CXC chemokine receptors on cultured FLS was studied at the messenger RNA (mRNA) level by reverse transcriptase-polymerase chain reaction and at the cell surface expression level by flow cytometry. Variations in cytosolic calcium influx induced by chemokine stimulation were assessed by flow cytometry on Fura Red-preloaded FLS. Two-compartment transwell chambers were used for FLS chemotaxis assays. Cell growth was measured by a fluorescence-based proliferation assay. Gelatinase and collagenase activities were determined by a fibril degradation assay and zymography.Results. FLS constitutively expressed the receptors CCR2, CCR5, CXCR3, and CXCR4, both at the cell surface and mRNA levels, but failed to express CCR3 and CCR6. Significant intracytosolic calcium influx was observed on FLS challenged with monocyte chemotactic protein 1 (MCP-1), stromal cell-derived factor la (SDF-1alpha), and interferon-inducible protein 10 (IP-10). Stimulation with MCP-1, SDF-1alpha, IP-10, and monokine induced by interferon-gamma enhanced the migration and proliferation of FLS. These chemokines, in addition to RANTES, increased in a dose- and time-dependent manner the gelatinase and collagenase activities in cell-free supernatants of cultured FLS. Interestingly, the chemokine-mediated up-regulation of MMP activities was significantly abrogated by the presence of anti-interleukin-1beta, but not anti-tumor necrosis factor alpha, blocking antibodies.Conclusion. These data suggest that through modulation of the migration, proliferation, and MNIP production by FLS, the chemokine system may play a more direct role in the destructive phase of RA than is currently suspected, and thus emphasize the relevance of chemokines and their receptors as potential therapeutic targets in this disease.