Han Yuli, Liu Changqing, Chen Shujing, Sun Huihui, Jia Zhaoyu, Shi Jiaxin, Wang Lirong, Du Kunze, Chang Yanxu
State Key Laboratory of Chinese Medicine Modernization, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China; State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
State Key Laboratory of Chinese Medicine Modernization, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
J Adv Res. 2025 Aug;74:609-620. doi: 10.1016/j.jare.2024.09.030. Epub 2024 Oct 5.
Rheumatoid arthritis (RA) is an autoimmune disease pathologically characterized by synovial inflammation. The abnormal activation of synoviocytes seems to accompany the progression of RA. The role and exact molecular mechanism in RA of columbianadin (CBN) which is a natural coumarin is still unclear.
The present research aimed to investigate the effect of vimentin on the abnormal growth characteristics of RA synoviocytes and the targeted regulatory role of CBN.
Cell migration and invasion were detected using the wound healing and transwell method. Mechanistically, the direct molecular targets of CBN were screened and identified by activity-based protein profiling. The expression of relevant proteins and mRNA in cells and mouse synovium was detected by western blotting and qRT-PCR. Changes in the degree of paw swelling and body weight of mice were recorded. H&E staining, toluidine blue staining, and micro-CT were used to visualize the degree of pathological damage in the ankle joints of mice. Small interfering RNA and plasmid overexpression of vimentin were used to observe their effects on MH7A cell proliferation, migration, apoptosis, and downstream molecular signaling.
The TNF-α-induced proliferation and migration of MH7A cells could be significantly repressed by CBN (25,50 μM), and the expression of apoptosis and autophagy-associated proteins could be modulated. Furthermore, CBN could directly bind to vimentin and inhibit its expression and function in synoviocytes, thereby ameliorating foot and paw swelling and joint damage in CIA mice. Silencing and overexpression of vimentin might be involved in developing RA synovial hyperplasia and invasive cartilage by activating VAV2 phosphorylation-mediated expression of Rac-1, which affects abnormal growth characteristics, such as synoviocyte invasion and migration.
CBN-targeted vimentin restrains the overactivation of RA synoviocytes thereby delaying the pathological process in CIA mice, which provides valuable targets and insights for understanding the pathological mechanisms of RA synovial hyperplasia.
类风湿关节炎（RA）是一种以滑膜炎症为病理特征的自身免疫性疾病。滑膜细胞的异常激活似乎伴随着RA的进展。天然香豆素类化合物哥伦比亚内酯（CBN）在RA中的作用及确切分子机制仍不清楚。
本研究旨在探讨波形蛋白对RA滑膜细胞异常生长特性的影响以及CBN的靶向调控作用。
采用划痕实验和Transwell法检测细胞迁移和侵袭能力。机制上，通过基于活性的蛋白质谱分析筛选和鉴定CBN的直接分子靶点。采用蛋白质免疫印迹法和qRT-PCR检测细胞及小鼠滑膜中相关蛋白和mRNA的表达。记录小鼠爪肿胀程度和体重变化。采用苏木精-伊红染色、甲苯胺蓝染色和显微CT观察小鼠踝关节病理损伤程度。利用波形蛋白的小干扰RNA和质粒过表达观察其对MH7A细胞增殖、迁移、凋亡及下游分子信号的影响。
CBN（25、50 μM）可显著抑制TNF-α诱导的MH7A细胞增殖和迁移，并可调节凋亡和自噬相关蛋白的表达。此外，CBN可直接与波形蛋白结合，抑制其在滑膜细胞中的表达和功能，从而改善胶原诱导性关节炎（CIA）小鼠的足爪肿胀和关节损伤。波形蛋白的沉默和过表达可能通过激活VAV2磷酸化介导的Rac-1表达参与RA滑膜增生和软骨侵袭的发生发展，进而影响滑膜细胞侵袭和迁移等异常生长特性。
CBN靶向波形蛋白可抑制RA滑膜细胞的过度激活，从而延缓CIA小鼠的病理进程，为理解RA滑膜增生的病理机制提供了有价值的靶点和见解。
