Wang Changyao, Zhang Han, Wang Xiangyu, Ma Xiao, Zhang Jian, Zhang Yongtao
Department of Orthopedics, The Affiliated Hospital of Qingdao University, Qingdao, 266000, Shandong, China.
Department of Traumatology, Fushan People's Hospital, Yantai, 265500, China.
Mol Cell Biochem. 2025 Mar;480(3):1669-1684. doi: 10.1007/s11010-024-05073-2. Epub 2024 Aug 7.
Rheumatoid arthritis (RA) is a chronic autoimmune disease that can cause destruction of cartilage and bone's extracellular matrix. Bromodomain 4 (BRD4), as a transcriptional and epigenetic regulator, plays a key role in cancer and inflammatory diseases. While, the role of BRD4 in bone destruction in RA has not been extensively reported. Our study aimed to investigate the effect of BRD4 on the bone destruction in RA and, further, its mechanism in the pathogenesis of the disease. In this study, receiving approval from the Ethical Committee of the Affiliated Hospital of Qingdao University, we evaluated synovial tissues from patients with RA and OA for BRD4 expression through advanced techniques such as immunohistochemistry, quantitative real-time PCR (qRT-PCR), and Western blotting. We employed a collagen-induced arthritis (CIA) mouse model to assess the therapeutic efficacy of the BRD4 inhibitor JQ1 on disease progression and bone destruction, supported by detailed clinical scoring and histological examinations. Further, in vitro osteoclastogenesis assays using RAW264.7 macrophages, facilitated by TRAP staining and resorption pit assays, provided insights into the mechanistic effects of JQ1 on osteoclast function. Statistical analysis was rigorously conducted using SPSS, applying Kruskal-Wallis, one-way ANOVA, and Student's t-tests to validate the data. In our study, we found that BRD4 expression significantly increased in the synovial tissues of RA patients and the ankle joints of CIA mice, with JQ1, a BRD4 inhibitor, effectively reducing inflammation, arthritis severity (p < 0.05), and bone erosion. Treatment with JQ1 not only improved bone mass and structural integrity in CIA mice but also downregulated osteoclast-related gene expression and the RANKL/RANK signaling pathway, indicating a suppression of osteolysis. Furthermore, in vitro assays demonstrated that JQ1 markedly inhibited osteoclast differentiation and function, underscoring the pivotal role of BRD4 in osteoclastogenesis and its potential as a target for therapeutic intervention in RA-induced bone destruction. Our study concludes that targeting BRD4 with the inhibitor JQ1 significantly mitigates inflammation and bone destruction in rheumatoid arthritis, suggesting that inhibition of BRD4 may be a potential therapeutic strategy for the treatment of bone destruction in RA.
类风湿关节炎(RA)是一种慢性自身免疫性疾病,可导致软骨和骨细胞外基质的破坏。溴结构域4(BRD4)作为一种转录和表观遗传调节因子,在癌症和炎症性疾病中起关键作用。然而,BRD4在类风湿关节炎骨破坏中的作用尚未得到广泛报道。我们的研究旨在探讨BRD4对类风湿关节炎骨破坏的影响及其在疾病发病机制中的作用机制。在本研究中,经青岛大学附属医院伦理委员会批准,我们通过免疫组织化学、定量实时PCR(qRT-PCR)和蛋白质印迹等先进技术评估类风湿关节炎患者和骨关节炎患者滑膜组织中BRD4的表达。我们采用胶原诱导性关节炎(CIA)小鼠模型,通过详细的临床评分和组织学检查,评估BRD4抑制剂JQ1对疾病进展和骨破坏的治疗效果。此外,使用RAW264.7巨噬细胞进行的体外破骨细胞生成试验,通过抗酒石酸酸性磷酸酶(TRAP)染色和吸收陷窝试验,深入了解了JQ1对破骨细胞功能的作用机制。使用SPSS进行严格的统计分析,应用Kruskal-Wallis检验、单因素方差分析和学生t检验来验证数据。在我们的研究中,我们发现类风湿关节炎患者滑膜组织和CIA小鼠踝关节中BRD4表达显著增加,BRD4抑制剂JQ1有效减轻炎症、关节炎严重程度(p<0.05)和骨侵蚀。用JQ¬1治疗不仅改善了CIA小鼠的骨量和结构完整性,还下调了破骨细胞相关基因表达和RANKL/RANK信号通路,表明抑制了骨吸收。此外,体外试验表明JQ1显著抑制破骨细胞分化和功能,强调了BRD4在破骨细胞生成中的关键作用及其作为类风湿关节炎诱导的骨破坏治疗干预靶点的潜力。我们的研究得出结论,用抑制剂JQ1靶向BRD4可显著减轻类风湿关节炎的炎症和骨破坏,表明抑制BRD4可能是治疗类风湿关节炎骨破坏的潜在治疗策略。