Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou Province 550025, China.
The 2nd Hospital Affiliated with Guizhou University of Chinese Traditional Medicine, Guiyang, Guizhou Province 550003, China.
Mediators Inflamm. 2023 Feb 8;2023:4814412. doi: 10.1155/2023/4814412. eCollection 2023.
Rheumatoid arthritis (RA) is a chronic progressive autoimmune disease characterized with high recurrence, high disability, poor prognosis, and long treatment cycles. Versus western medicine, traditional Chinese medicine has the traits of definite efficacy, low toxicity, and side effects in the treatment of RA. Moreover, traditional Chinese medicine also has the advantages of multiple targets, multiple links, and multiple approaches. This study was committed to exploring the effect of Jinwujiangu prescription on peripheral blood osteoclasts in those patients with RA and relevant molecular mechanisms. We first identified 159 common targets by online pharmacology, and there were correlations among these targets; besides, the main signaling pathways involved were inclusive TNF signaling pathway, rheumatoid arthritis, IL-17 signaling pathway, NF-kappa B signaling pathway, Toll-like receptor signaling pathway, etc. Through experimental verification, we found that PBMC cells extracted from human peripheral blood could be successfully induced into osteoclasts, and Jinwujiangu prescription inhibited the generation of osteoclasts from PBMCs of RA patients. CCK-8 and flow cytometry showed that osteoclast viability was significantly decreased and osteoclast apoptosis was significantly increased in the HIF-1 interference group; low-, medium-, and high-dose Jinwujiangu prescription groups; sinapine group; and hydroxychloroquine control group. Moreover, Jinwujiangu prescription and sinapine could inhibit the production of cytokines in peripheral blood osteoclasts and inhibit autophagy in RA patients. The expression level of mTOR was significantly increased in both Jinwu middle- and high-dose groups. In conclusion, this study demonstrated that sinapine, the active target in Jinwujiangu prescription, can act as a HIF-1 inhibitor; activate the mTOR pathway; downregulate the level of autophagy rate, ATG5, beclin-1, and LC3 expression; and inhibit the occurrence of autophagy. The trial registration number of the study is KYW2021010.
类风湿关节炎（RA）是一种慢性进行性自身免疫性疾病，具有高复发、高残疾、预后差和治疗周期长的特点。与西药相比，中药在治疗 RA 方面具有疗效确切、毒性低、副作用小的特点。此外，中药还具有多靶点、多环节、多途径的优势。本研究旨在探讨金乌健骨方对 RA 患者外周血破骨细胞的作用及相关分子机制。我们首先通过网络药理学鉴定出 159 个共同靶点，这些靶点之间存在相关性；此外，涉及的主要信号通路包括 TNF 信号通路、类风湿关节炎、IL-17 信号通路、NF-kappa B 信号通路、Toll 样受体信号通路等。通过实验验证，我们发现可以从人外周血中提取 PBMC 细胞，并成功诱导其分化为破骨细胞，金乌健骨方可抑制 RA 患者 PBMC 来源的破骨细胞的生成。CCK-8 和流式细胞术结果显示，HIF-1 干扰组、低、中、高剂量金乌健骨方组、白芥子组和羟氯喹对照组破骨细胞活力显著降低，破骨细胞凋亡显著增加。此外，金乌健骨方和白芥子均可抑制外周血破骨细胞细胞因子的产生，并抑制 RA 患者的自噬。金乌中、高剂量组 mTOR 表达水平明显升高。综上所述，本研究表明金乌健骨方中的活性成分白芥子可作为 HIF-1 抑制剂，激活 mTOR 通路，下调自噬率、ATG5、beclin-1 和 LC3 表达水平，抑制自噬的发生。该研究的注册号为 KYW2021010。
