Department of Immunology and Rheumatology, Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital), Tongji Medical College, Huazhong University of Science and Technology, WuHan, China.
Biosci Rep. 2020 Aug 28;40(8). doi: 10.1042/BSR20201273.
Rheumatoid arthritis (RA) is a frequent autoimmune disease. Emerging evidence indicated that ZNFX1 antisense RNA1 (ZFAS1) participates in the physiological and pathological processes in RA. However, knowledge of ZFAS1 in RA is limited, the potential work pathway of ZFAS1 needs to be further investigated.
Levels of ZFAS1, microRNA (miR)-2682-5p, and ADAM metallopeptidase with thrombospondin type 1 motif 9 (ADAMTS9) were estimated using quantitative real-time polymerase chain reaction (qRT-PCR) assay. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was conducted to explore the ability of cell proliferation in fibroblast-like synoviocytes (FLS-RA). Cell apoptosis was measured via flow cytometry. Also, levels of ADAMTS9, apoptosis-related proteins, cleaved-caspase-3 (active large subunit), and autophagy-related proteins were identified adopting Western blot. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the productions of inflammatory cytokines. Beside, the interrelation between miR-2682-5p and ZFAS1 or ADAMTS9 was verified utilizing dual-luciferase reporter assay.
High levels of ZFAS1 and ADAMTS9, and a low level of miR-2682-5p were observed in RA synovial tissues and FLS-RA. Knockdown of ZFAS1 led to the curbs of cell proliferation, inflammation, autophagy, and boost apoptosis in FLS-RA, while these effects were abolished via regaining miR-2682-5p inhibition. Additionally, the influence of miR-2682-5p on cell phenotypes and inflammatory response were eliminated by ADAMTS9 up-regulation in FLS-RA. Mechanically, ZFAS1 exerted its role through miR-2682-5p/ADAMTS9 axis in RA.
ZFAS1/miR-2682-5p/ADAMTS9 axis could modulate the cell behaviors, inflammatory response in FLS-RA, might provide a potential therapeutic target for RA treatment.
类风湿关节炎（RA）是一种常见的自身免疫性疾病。新出现的证据表明，锌指蛋白 X1 反义 RNA1（ZFAS1）参与 RA 的生理和病理过程。然而，ZFAS1 在 RA 中的知识有限，需要进一步研究其潜在的工作途径。
采用实时定量聚合酶链反应（qRT-PCR）法检测 ZFAS1、微小 RNA（miR）-2682-5p 和含有血小板反应蛋白 1 型基序的解金属蛋白酶 9（ADAMTS9）的水平。采用 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）法检测成纤维样滑膜细胞（FLS-RA）的增殖能力。通过流式细胞术检测细胞凋亡。此外，采用 Western blot 法检测 ADAMTS9、凋亡相关蛋白、裂解型半胱天冬酶-3（活性大片段）和自噬相关蛋白的水平。采用酶联免疫吸附试验（ELISA）法测定炎症细胞因子的产生。此外，利用双荧光素酶报告基因实验验证 miR-2682-5p 与 ZFAS1 或 ADAMTS9 之间的相互关系。
RA 滑膜组织和 FLS-RA 中 ZFAS1 和 ADAMTS9 水平升高，miR-2682-5p 水平降低。ZFAS1 敲低可抑制 FLS-RA 细胞增殖、炎症、自噬，并促进细胞凋亡，而恢复 miR-2682-5p 抑制则可消除这些作用。此外，在 FLS-RA 中上调 ADAMTS9 可消除 miR-2682-5p 对细胞表型和炎症反应的影响。机制上，ZFAS1 通过 RA 中的 miR-2682-5p/ADAMTS9 轴发挥作用。
ZFAS1/miR-2682-5p/ADAMTS9 轴可调节 FLS-RA 中的细胞行为和炎症反应，可能为 RA 治疗提供潜在的治疗靶点。
