Proline-rich tyrosine kinase 2 and Src kinase signaling transduce monosodium urate crystal-induced nitric oxide production and matrix metalloproteinase 3 expression in chondrocytes
Proline-rich tyrosine kinase 2 and Src kinase signaling transduce monosodium urate crystal-induced nitric oxide production and matrix metalloproteinase 3 expression in chondrocytes
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DOI：
10.1002/art.11486
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发表时间：
2004-01-01
影响因子：
--
通讯作者：
Terkeltaub, R
中科院分区：
文献类型：
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作者：
Liu, R;Lioté, F;Terkeltaub, R
目标。关节内一水尿酸钠(MSU)晶体的沉积可促进软骨和骨的侵蚀。因此，本研究的目的是确定MSU晶体对软骨细胞的刺激作用。观察MSU对培养软骨细胞一氧化氮(NO)释放、诱导型一氧化氮合酶(INOS)和基质金属蛋白酶3(MMP3)表达的影响。应用选择性药物抑制剂和转染激酶突变体的方法，研究了MSU诱导的特异性蛋白激酶(p38、Src和粘着斑激酶[FAK]家族成员富含脯氨酸的酪氨酸激酶2[PYK-2]和FAK)的功能信号。通过使用IL-1受体拮抗剂，MSU以p38依赖的方式诱导软骨细胞表达基质金属蛋白酶-3和诱导型一氧化氮合酶，并释放一氧化氮，而不需要白细胞介素1(IL-1)。MSU诱导PYK-2和FAK及其接头蛋白Paxlin和相互作用蛋白c-Src的快速酪氨酸磷酸化。PYK-2和c-Src信号均介导MSU诱导的p38MAPK激活。PYK-2和c-Src信号在MSU诱导的NO产生和基质金属蛋白酶-3的表达中起主要作用。但是，尽管观察到FAK的磷酸化，但在平行实验中，选择性药理FAK抑制剂和FAK显性-阴性突变体都未能阻止MSU诱导的NO释放或基质金属蛋白酶-3的表达。在软骨细胞中，MSU晶体激活了通常由黏附受体激活的信号激酶级联，包括上游的Src和FAK家族的激活以及下游的p38激活。在这一级联反应中，PYK-2、Src和p38激酶转导MSU诱导的NO产生和MMP3的表达。我们的结果确定了PYK-2和c-Src是潜在的治疗干预慢性痛风软骨退化的新位点。
Objective. Articular deposition of monosodium urate monohydrate (MSU) crystals may promote cartilage and bone erosion. Therefore, the aim of this study was to determine how MSU crystals stimulate chondrocytes.Methods. Nitric oxide (NO) release, and expression of inducible nitric oxide synthase (iNOS) and matrix metalloproteinase 3 (MMP-3) were assessed in cultured chondrocytes treated with MSU. MSU-induced functional signaling by specific protein kinases (p38, Src, and the focal adhesion kinase [FAK] family members proline-rich tyrosine kinase 2 [Pyk-2] and FAK) was also examined using selective pharmacologic inhibitors and transfection of kinase mutants.Results. MSU induced MMP-3 and iNOS expression and NO release in chondrocytes in a p38-dependent manner that did not require interleukin-1 (IL-1), as demonstrated by using IL-1 receptor antagonist. MSU induced rapid tyrosine phosphorylation of Pyk-2 and FAK, their adaptor protein paxillin, and interacting kinase c-Src. Pyk-2 and c-Src signaling both mediated p38 MAPK activation in response to MSU. Pyk-2 and c-Src signaling played a major role in transducing MSU-induced NO production and MMP-3 expression. But, despite the observed FAK phosphorylation, a selective pharmacologic FAK inhibitor and a FAK dominant-negative mutant both failed to block MSU-induced NO release or MMP-3 expression in parallel experiments.Conclusion. In chondrocytes, MSU crystals activate a signaling kinase cascade typically employed by adhesion receptors that involves upstream Src and FAK family activation and downstream p38 activation. In this cascade, Pyk-2, Src, and p38 kinases transduce MSU-induced NO production and MMP-3 expression. Our results identify Pyk-2 and c-Src as novel sites for potential therapeutic intervention in cartilage degradation in chronic gout.