Iguratimod Inhibits the Aggressiveness of Rheumatoid Fibroblast-Like Synoviocytes
Iguratimod Inhibits the Aggressiveness of Rheumatoid Fibroblast-Like Synoviocytes
复制标题
艾拉莫德抑制类风湿成纤维细胞样滑膜细胞的攻击性
DOI：
10.1155/2019/6929286
复制
发表时间：
2019-11-14
影响因子：
4.1
通讯作者：
Chen, Weiqian
中科院分区：
文献类型：
--
作者：
Lin, Jin;Yu, Ye;Chen, Weiqian
目的Iguratimod是一种治疗类风湿关节炎的新型抗风湿药物，已在中国和日本获得批准。在这里，我们的目的是发现iguratimod是否可以抑制类风湿成纤维细胞样滑膜细胞（RA-FLSs）的侵袭行为并促进细胞凋亡。方法采用5-乙基-2 ' -脱氧尿苷试验和细胞计数试剂盒-8检测RA-FLSs的增殖情况。通过创面试验和transwell实验测定迁移和侵袭。流式细胞术检测细胞凋亡。采用定量PCR和酶联免疫吸附法检测RA-FLSs中基质金属蛋白酶（MMPs）和促炎细胞因子mRNA的表达。为了深入了解分子信号机制，我们通过细胞热移测定（CETSA）和western blot确定了iguratimod对丝裂原活化蛋白激酶（MAPK）信号通路激活的影响。结果Iguratimod可显著降低RA-FLSs的体外增殖、迁移和侵袭能力，且呈剂量依赖性。iguratimod治疗后，MMP-1、MMP-3、MMP-9、白细胞介素-6 （IL-6）和单核细胞趋化蛋白-1 mRNA和蛋白水平均降低。此外，肿瘤坏死因子-α- (TNF-α-)诱导的磷酸化c-Jun n -末端激酶（JNK）和P38 MAPK的表达被iguratimod抑制。此外，iguratimod促进RA-FLSs的凋亡。最重要的是，通过CETSA实验，iguratimod被证明直接与JNK和P38蛋白相互作用。此外，激活转录因子2 (ATF-2)， JNK和P38的底物，被iguratimod抑制。结论iguratimod对RA的治疗作用可能部分是由于靶向RA- flss的侵袭性行为和凋亡。
Objective Iguratimod, a novel disease-modifying anti-rheumatic drug for the treatment of rheumatoid arthritis, has been approved in China and Japan. Here, we aimed to find whether iguratimod can inhibit the aggressive behavior and promote apoptosis of rheumatoid fibroblast-like synoviocytes (RA-FLSs). Methods The proliferation of RA-FLSs was assessed by 5-ethynyl-2′-deoxyuridine test and Cell Counting Kit-8. Migration and invasion were determined by the wound test and a transwell assay. Apoptosis was tested by flow cytometry. The mRNA expression of matrix metalloproteinases (MMPs) and proinflammatory cytokines in RA-FLSs were measured by quantitative PCR and ELISA. To gain insight into the molecular signaling mechanisms, we determined the effect of iguratimod on the activation of mitogen-activated protein kinases (MAPK) signaling pathways by the cellular thermal shift assay (CETSA) and western blot. Results Iguratimod treatment significantly reduced the proliferation, migration, and invasive capacities of RA-FLSs in a dose-dependent manner in vitro. MMP-1, MMP-3, MMP-9, Interleukin-6 (IL-6), and monocyte chemoattractant protein-1 mRNA and protein levels were all decreased after treatment with iguratimod. Furthermore, tumor necrosis factor-alpha- (TNF-α-) induced expression of phosphorylated c-Jun N-terminal kinases (JNK) and P38 MAPK were inhibited by iguratimod. Additionally, iguratimod promoted the apoptosis of RA-FLSs. Most importantly, iguratimod was shown to directly interact with JNK and P38 protein by CETSA assay. Moreover, activating transcription factor 2 (ATF-2), a substrate of both JNK and P38, was suppressed by iguratimod. Conclusions Our findings suggested that the therapeutic effects of iguratimod on RA might be, in part, due to targeting the aggressive behavior and apoptosis of RA-FLSs.