We previously reported that penta-acetyl geniposide ((Ac)₅GP, an active derivative of geniposide) showed anti-arthritic effect on adjuvant-induced arthritis (AIA) rats by promoting the apoptosis of AIA fibroblast-like synoviocyte (FLS). This study aimed to demonstrate the effects of (Ac)₅GP on migration, invasion, and inflammation of TNF-α-stimulated rheumatoid arthritis (RA) FLS (MH7A cell) and to explore the involved mechanisms. MTT assay was used to determine the applied non-cytotoxic doses of (Ac)₅GP (12.5, 25, 50 μM) in vitro. Results of wound-healing, transwell, and phalloidin staining assays indicated that (Ac)₅GP reduced the migration, invasion, and F-actin cytoskeletal reorganization of TNF-α-stimulated MH7A. Results of ELISA and western blot assays confirmed that (Ac)₅GP reduced TNF-α-induced production of pro-inflammatory cytokines (like IL-1β, IL-6, IL-8) and matrix metalloproteinases (MMPs, such as MMP-2 and MMP-9). Moreover, (Ac)₅GP inhibited TNF-α-induced activation of Wnt/β-catenin pathway, evidenced by reducing the protein levels of Wnt1, p-GSK-3β (Ser9), and β-catenin and preventing β-catenin nuclear translocation. Importantly, the combination of XAV939 (an inhibitor of Wnt/β-catenin) promoted the actions of (Ac)₅GP on TNF-α-induced migration, invasion, and inflammation, further revealing the involvement of Wnt/β-catenin pathway underlying the therapeutic effects of (Ac)₅GP on TNF-α-stimulated MH7A. In vivo, (Ac)₅GP relieved the progression and severity of rat collagen-induced arthritis, related to reducing the levels of IL-1β, IL-6, IL-8, MMP-2, and MMP-9 as well as inhibiting Wnt/β-catenin pathway in synovial tissues. Collectively, (Ac)₅GP could suppress TNF-α-induced migration, invasion, and inflammation in RA FLS involving Wnt/β-catenin pathway and (Ac)₅GP might be as a candidate agent for RA treatment.

我们之前报道了五乙酰栀子苷（（Ac）₅ GP，栀子苷的活性衍生物）通过促进AIA成纤维细胞样滑膜细胞（FLS）的凋亡对佐剂性关节炎（AIA）大鼠显示出抗关节炎作用。本研究旨在证明 (Ac) ₅ GP 对 TNF-α 刺激的类风湿性关节炎 (RA) FLS (MH7A 细胞) 的迁移、侵袭和炎症的影响，并探讨其相关机制。_{MTT测定用于确定体外应用的(Ac) 5} GP (12.5, 25, 50 μM)的非细胞毒性剂量。伤口愈合、transwell 和鬼笔环肽染色试验的结果表明 (Ac) ₅GP 减少了 TNF-α 刺激的 MH7A 的迁移、侵袭和 F-肌动蛋白细胞骨架重组。ELISA 和蛋白质印迹分析的结果证实，(Ac) ₅ GP 可减少 TNF-α 诱导的促炎细胞因子（如 IL-1β、IL-6、IL-8）和基质金属蛋白酶（MMP，如 MMP- 2和MMP-9）。此外，(Ac) ₅ GP 抑制 TNF-α 诱导的 Wnt/β-catenin 通路激活，这可以通过降低 Wnt1、p-GSK-3β (Ser9) 和 β-catenin 的蛋白质水平并阻止 β-catenin 核易位。重要的是，XAV939（Wnt/β-catenin 抑制剂）的组合促进了 (Ac) _5的作用GP 对 TNF-α 诱导的迁移、侵袭和炎症的影响，进一步揭示了 Wnt/β-catenin 通路参与了 (Ac) ₅ GP 对 TNF-α 刺激的 MH7A 的治疗作用。在体内，(Ac) ₅ GP 减轻了大鼠胶原诱导的关节炎的进展和严重程度，这与降低 IL-1β、IL-6、IL-8、MMP-2 和 MMP-9 的水平以及抑制滑膜组织中的 Wnt/β-catenin 通路。总的来说，(Ac) ₅ GP 可以抑制 TNF-α 诱导的涉及 Wnt/β-catenin 通路的 RA FLS 中的迁移、侵袭和炎症，并且 (Ac) ₅ GP 可能是 RA 治疗的候选药物。