Role of p21-activated kinase 1 in regulating the migration and invasion of fibroblast-like synoviocytes from rheumatoid arthritis patients
Role of p21-activated kinase 1 in regulating the migration and invasion of fibroblast-like synoviocytes from rheumatoid arthritis patients
复制标题
p21激活激酶1在调节类风湿性关节炎成纤维样滑膜细胞迁移和侵袭中的作用
DOI：
10.1093/rheumatology/kes031
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发表时间：
2012-07-01
期刊：
影响因子：
5.5
通讯作者：
Xu, Hanshi
中科院分区：
文献类型：
--
作者：
Fu, Di;Yang, Yanlong;Xu, Hanshi
目标。目的探讨p21活化激酶1 （PAK1）在RA成纤维细胞样滑膜细胞（FLS）迁移、侵袭及MMP表达调控中的作用。采用Boyden室法测定RA FLS在体外的迁移和侵袭。在严重联合免疫缺陷（SCID）小鼠体内RA共植入模型中检测到RA FLS对软骨的侵袭。western blotting检测PAK1和MT1-MMP的表达。ELISA法检测mmp的产生及活性。在RA患者离体滑膜细胞中磷酸化PAK1 （p-PAK1）蛋白表达增加。IL-1或tnf - α刺激可上调p-PAK1的表达。转染PAK1显性阴性突变体（dnPAK1）抑制PAK1可减少RA FLS的体外迁移和侵袭。在SCID小鼠模型中，体内转染dnPAK1可减弱RA FLS对软骨的侵袭。PAK1调节IL-1 β诱导的MMP-13和MT1-MMP的产生和活性。抑制MMP-13或MT1-MMP活性也可减少RA FLS侵袭。此外，转染dnPAK1可抑制c-Jun n -末端激酶（JNK）的激活，但不影响细胞外信号调节激酶和p38的活性。化学抑制剂抑制JNK活性可显著降低MMP-13和mt1 - mmp的迁移、侵袭和产生。PAK1在RA FLS中调控MMPs的迁移、侵袭、产生和活性，并通过JNK通路介导。这提示了一种针对PAK1的新策略来防止RA的联合破坏。
Objective. To investigate the role of p21-activated kinase 1 (PAK1) in regulating migration, invasion and MMP expression in RA fibroblast-like synoviocytes (FLS).Methods. RA FLS migration and invasion in vitro were measured by the Boyden chamber method. Invasion of RA FLS into cartilage was detected in the severe combined immunodeficiency (SCID) mouse co-implantation model of RA in vivo. PAK1 and MT1-MMP expression were examined by western blotting. ELISA was used to measure the production and activity of MMPs.Results. Phosphorylated PAK1 (p-PAK1) protein expression was increased in ex vivo synovial membrane cells from RA patients. Stimulation with IL-1 beta or TNF-alpha up-regulated p-PAK1 expression. Inhibition of PAK1 by transfection with dominant negative PAK1 mutant (dnPAK1) reduced in vitro migration and invasion of RA FLS. In the SCID mouse model, RA FLS invasion into cartilage was attenuated by transfection with dnPAK1 in vivo. PAK1 regulated IL-1 beta-induced production and activity of MMP-13 and MT1-MMP. Inhibition of MMP-13 or MT1-MMP activity also reduced RA FLS invasion. Furthermore, dnPAK1 transfection inhibited c-Jun N-terminal kinase (JNK) activation, but did not affect the activities of extracellular signal-regulated kinases and p38. Inhibition of the JNK activity by chemical inhibitor significantly reduced the migration, invasion and production of MMP-13 and MT1-MMP.Conclusion. PAK1 plays an important role in regulating the migration, invasion and production and activity of MMPs in RA FLS, which is mediated by the JNK pathway. This suggests a novel strategy targeting PAK1 to prevent joint destruction of RA.