Abstract

Rheumatoid arthritis (RA) is considered to be a chronic autoimmune disease, pathogenesis of RA is complex and effective treatments for RA is still lacking. Previous studies found that microRNAs (miRNAs) play important roles in the pathogenesis of RA, and miR-223-3p is considered to be one of the possible biomarkers of RA. Recent studies have revealed that icariin alleviates RA in murine models, but the underlying mechanism needs to be further investigated. MiR-223-3p expression levels in fibroblast-like synoviocyte (RA-FLS) and patients with RA were quantified by qRT-PCR, cell proliferation was analyzed by CCK-8 and BrdU assay. Cell apoptosis was assessed by flow cytometry and western blotting. TNF-α, IL-1β and IL-6 concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Dual luminescence-based reporter gene assay was conducted to confirm the possible interaction between miR-223-3p and NLRP3. Icariin inhibits proliferation and inflammation cytokines secretion, promotes apoptosis of RA-FLS cells and upregulated the expression of miR-223-3p. MiR-223-3p targets to 3’-UTR of NRLP3 and regulates its expression. MiR-223-3p inhibitor reversed the effect of icariin on RA-FLS cells function. Additionally, anti-RA activity of icariin was restored by NLRP3 inhibitor MCC950 in miR-223-3p knockdown RA-FLS cells. Icariin inhibits proliferation and inflammation, promotes apoptosis of RA-FLS cells by regulating miR-223-3p/NLRP3 signalling, which may serve as a potential therapeutic target to alleviate RA.

摘要

类风湿性关节炎（RA）被认为是一种慢性自身免疫性疾病，其发病机制复杂，目前尚缺乏有效的治疗方法。以往的研究发现microRNAs（miRNAs）在RA的发病机制中起重要作用，miR-223-3p被认为是RA可能的生物标志物之一。最近的研究表明淫羊藿苷可减轻小鼠模型中的 RA，但其潜在机制需要进一步研究。通过qRT-PCR定量成纤维细胞样滑膜细胞（RA-FLS）和RA患者中的MiR-223-3p表达水平，通过CCK-8和BrdU测定分析细胞增殖。通过流式细胞术和蛋白质印迹评估细胞凋亡。TNF-α、IL-1β 和 IL-6 浓度通过酶联免疫吸附测定 (ELISA) 进行测量。进行基于双发光的报告基因测定以确认 miR-223-3p 和 NLRP3 之间可能的相互作用。淫羊藿苷抑制增殖和炎症细胞因子分泌，促进 RA-FLS 细胞凋亡并上调 miR-223-3p 的表达。MiR-223-3p 靶向 NRLP3 的 3'-UTR 并调节其表达。MiR-223-3p 抑制剂逆转淫羊藿苷对 RA-FLS 细胞功能的影响。此外，在 miR-223-3p 敲低 RA-FLS 细胞中，NLRP3 抑制剂 MCC950 恢复了淫羊藿苷的抗 RA 活性。淫羊藿苷通过调节 miR-223-3p/NLRP3 信号传导抑制增殖和炎症，促进 RA-FLS 细胞的凋亡，这可能是缓解 RA 的潜在治疗靶点。淫羊藿苷抑制增殖和炎症细胞因子分泌，促进 RA-FLS 细胞凋亡并上调 miR-223-3p 的表达。MiR-223-3p 靶向 NRLP3 的 3'-UTR 并调节其表达。MiR-223-3p 抑制剂逆转淫羊藿苷对 RA-FLS 细胞功能的影响。此外，在 miR-223-3p 敲低 RA-FLS 细胞中，NLRP3 抑制剂 MCC950 恢复了淫羊藿苷的抗 RA 活性。淫羊藿苷通过调节 miR-223-3p/NLRP3 信号传导抑制增殖和炎症，促进 RA-FLS 细胞的凋亡，这可能是缓解 RA 的潜在治疗靶点。淫羊藿苷抑制增殖和炎症细胞因子分泌，促进 RA-FLS 细胞凋亡并上调 miR-223-3p 的表达。MiR-223-3p 靶向 NRLP3 的 3'-UTR 并调节其表达。MiR-223-3p 抑制剂逆转淫羊藿苷对 RA-FLS 细胞功能的影响。此外，在 miR-223-3p 敲低 RA-FLS 细胞中，NLRP3 抑制剂 MCC950 恢复了淫羊藿苷的抗 RA 活性。淫羊藿苷通过调节 miR-223-3p/NLRP3 信号传导抑制增殖和炎症，促进 RA-FLS 细胞的凋亡，这可能是缓解 RA 的潜在治疗靶点。在 miR-223-3p 敲低 RA-FLS 细胞中，NLRP3 抑制剂 MCC950 恢复淫羊藿苷的抗 RA 活性。淫羊藿苷通过调节 miR-223-3p/NLRP3 信号传导抑制增殖和炎症，促进 RA-FLS 细胞的凋亡，这可能是缓解 RA 的潜在治疗靶点。在 miR-223-3p 敲低 RA-FLS 细胞中，NLRP3 抑制剂 MCC950 恢复淫羊藿苷的抗 RA 活性。淫羊藿苷通过调节 miR-223-3p/NLRP3 信号传导抑制增殖和炎症，促进 RA-FLS 细胞的凋亡，这可能是缓解 RA 的潜在治疗靶点。