Activity of fibroblast-like synoviocytes in rheumatoid arthritis was impaired by dickkopf-1 targeting siRNA
Activity of fibroblast-like synoviocytes in rheumatoid arthritis was impaired by dickkopf-1 targeting siRNA
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dickkopf-1 靶向 siRNA 损害类风湿关节炎中成纤维细胞样滑膜细胞的活性
DOI：
10.1097/cm9.0000000000000697
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发表时间：
2020-03-20
影响因子：
6.1
通讯作者：
Li Zhan-Guo
中科院分区：
文献类型：
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作者：
Liu Yan-Ying;Wang Shi-Yao;Li Zhan-Guo
摘要背景纤维母细胞样滑膜细胞（FLSs）是滑膜关节的常驻间充质细胞，在类风湿关节炎（RA）的发病过程中起重要作用。Dickkopf-1 （DKK-1）被认为是炎症性关节炎中骨重塑的主要调控因子。在这里，我们研究了针对DKK-1的小干扰rna （sirna）对RA衍生的FLSs活性的潜在损害。方法将靶向DKK-1的sirna转染RA患者的FLSs。采用酶联免疫吸附法（ELISA）检测细胞培养上清中白细胞介素(IL)-1β、IL-6、IL-8、基质金属蛋白酶（MMP） 2、MMP3、MMP9、转化生长因子(TGF)-β1、TGF-β2和单核细胞趋化蛋白(MCP)-1水平。采用侵袭法和3H掺入法分别研究靶向DKK-1的sirna对FLSs侵袭和细胞增殖的影响。Western blotting检测FLSs中核因子(NF)-κB、白细胞介素-1受体相关激酶（IRAK）1、细胞外调节蛋白激酶（ERK）1、Jun n-末端激酶（JNK）和β-catenin的表达。结果DKK-1靶向sirna抑制了FLSs中DKK-1的表达（P < 0.01）。sirna诱导FLSs中IL-6、IL-8、MMP2、MMP3和MMP9水平较对照组显著降低（P < 0.05）。DKK-1靶向sirna抑制FLSs的增殖和侵袭（P < 0.05）。FLSs中促炎信号的重要分子，包括IRAK1和ERK1，通过抑制DKK-1而降低。相反，Wnt信号通路的关键下游分子β-catenin增加。结论通过抑制DKK-1，我们可以抑制RA源FLSs的增殖、侵袭和促炎细胞因子的分泌，这可能是由ERK或IRAK-1信号通路介导的。这些数据表明，DKK-1沉默的应用可能是一种潜在的治疗RA的方法。
Abstract Background Fibroblast-like synoviocytes (FLSs), resident mesenchymal cells of synovial joints, play an important role in the pathogenesis of rheumatoid arthritis (RA). Dickkopf-1 (DKK-1) has been proposed to be a master regulator of bone remodeling in inflammatory arthritis. Here, potential impairation on the activity of FLSs derived from RA to small interfering RNAs (siRNAs) targeting DKK-1 was investigated. Methods siRNAs targeting DKK-1 were transfected into FLSs of patients with RA. Interleukin (IL)-1β, IL-6, IL-8, matrix metalloproteinase (MMP) 2, MMP3, MMP9, transforming growth factor (TGF)-β1, TGF-β2 and monocyte chemoattractant protein (MCP)-1 levels in the cell culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Invasion assay and 3H incorporation assay were utilized to investigate the effects of siRNAs targeting DKK-1 on FLSs invasion and cell proliferation, respectively. Western blotting was performed to analyze the expression of nuclear factor (NF)-κB, interleukin-1 receptor-associated kinase (IRAK)1, extracellular regulated protein kinases (ERK)1, Jun N-terminal kinase (JNK) and β-catenin in FLSs. Results DKK-1 targeting siRNAs inhibited the expression of DKK-1 in FLSs (P < 0.01). siRNAs induced a significant reduction of the levels of IL-6, IL-8, MMP2, MMP3 and MMP9 in FLSs compared to the control group (P < 0.05). DKK-1 targeting siRNAs inhibited the proliferation and invasion of FLSs (P < 0.05). Important molecules of pro-inflammatory signaling in FLSs, including IRAK1 and ERK1, were decreased by the inhibition of DKK-1 in FLSs. In contrast, β-catenin, a pivotal downstream molecule of the Wnt signaling pathway was increased. Conclusions By inhibiting DKK-1, we were able to inhibit the proliferation, invasion and pro-inflammatory cytokine secretion of FLSs derived from RA, which was mediated by the ERK or the IRAK-1 signaling pathway. These data indicate the application of DKK-1 silencing could be a potential therapeutic approach to RA.