基于miR-125b探讨化瘀强肾通痹方干预NF-kB信号通路在类风湿关节炎的抗炎作用

Rheumatoid arthritis (RA) is an incurable disease of complicated pathogenesis. Chinese medicine (TCM) has certain advantages in improving RA efficiency and stability. Our team focused on treating RA by promoting blood circulation and removing blood stasis for more than 20 years. Based on the clinical experience and research results, correlated the academic idea that “kidney deficiency and blood stasis of RA”, combined with the academic thought of Chinese medicine master Zhu Liangchun and Jiao Shude in the treatment of Wangbi's, the Huayu Qiangshen Tongbi Decoction (HQT Decoction) was created. The efficacy, safety and stability of HQT Decoction and the combination of HQT Decoctionand MTX have been confirmed through the study of the clinical effect and the mechanism of animal pharmacodynamics. However, as the traditional Chinese medicine prescription, the way of how to exert its effect is always difficult to clarify, limiting its popularization and application and long-term development. MiR-125b is down-regulated in RA, which can activate CK2/NF-κB pathways, and ultimately promotes inflammation in RA. Therefore, we hypothesize that HQT Decoctionand can suppress the activity of CK2/NF-κB pathways by increasing the expression of miR-125b. This study is seek to identify the mechanism of HQT Decoction by animal and cell experiments. This study will to some extent reveal the scientific connotation the effect of HQT Decoctionand in the future.

类风湿关节炎（rheumatoid arthritis，RA）病机复杂，无法治愈，中医药在提高RA治疗有效率及稳定病情方面有一定优势。本项目组根据20多年“活血化瘀法”治疗RA的临床经验和研究成果，基于RA肾虚血瘀的病机认识，结合中医经典与国医大师朱良春和焦树德治疗尪痹的学术思想，提炼、创制“化瘀强肾通痹方”。前期已通过临床疗效以及动物药效机制研究肯定了化瘀强肾通痹方及其联合MTX的中西医结合方案的有效性，安全性和稳定性。然而化瘀强肾通痹方与其他中药复方一样，其发挥疗效的途径是如何始终难以阐明，限制其推广运用和长远发展。miR-125b在RA中低表达，激活CK2/NF-κB的活性，介导RA的炎症反应。本研究提出“化瘀强肾通痹方通过促进miR-125b的表达来抑制CK2/NF-κB信号通路“的科学假说，借助动物实验和细胞实验，多层面系统的探讨化瘀强肾通痹方治疗RA的物质基础，揭示其科学内涵。

类风湿关节炎（rheumatoid arthritis，RA）病机复杂，无法治愈，中医药在提高RA治疗有效率及稳定病情方面有一定优势。传统中药复方化瘀强肾通痹方对RA具有一定的疗效，但是其具体的分子机制尚不清楚。在本研究中，我们利用LPS处理的人类风湿成纤维样滑膜细胞（fibroblast-like synoviocytes，FLSs）MH7A细胞和胶原诱导的关节炎（CIA）大鼠作为体外和体内模型，研究化瘀强肾通痹方干预miR-125b介导RA炎症的机制。通过本研究，我们阐明了化瘀强肾通痹方可以通过下调TNF-α、IL-1β和IL-6等细胞因子的表达有效抑制RA炎症，研究同时证实了化瘀强肾通痹方能上调miR-125b的表达。利用TargetScan预测酪蛋白激酶2（CK2）是miR-125b的潜在靶点，荧光素酶报告基因检测发现在MH7A细胞中miR-125b可降低CK2的表达。miR-125b可抑制LPS诱导的（NF-kappa-B）NF-κB的激活，而NF-κB是CK2的下游靶点。NF-κB抑制剂PDTC和NF-κB抑制剂α（IkB-α）可以增强miR-125b对TNF-α、IL-1β和IL-6表达的抑制作用。另外，本研究证实了化瘀强肾通痹方通过上调miR-125b水平靶向抑制CK2表达来阻止NF-κB诱导的炎症反应，从而缓解RA的炎症反应。本研究结果将有助于进一步研究化瘀强肾通痹方对RA的作用机制，并为RA的治疗提供新的靶点。