Shi Mei-Feng, Liu Xiao-Bao, Ma Xiao-Na, Feng Wei, Zhang Yi-Fang, Lin Chang-Song, Liu Qing-Ping, Xu Qiang
State Key Laboratory of Traditional Chinese Medicine Syndrome, The First Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405, China; Department of Rheumatology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
State Key Laboratory of Traditional Chinese Medicine Syndrome, The First Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405, China; Department of Rheumatology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, China; Guangdong Clinical Research Academy of Chinese Medicine, Guangzhou 510405, China.
Int Immunopharmacol. 2025 Mar 26;150:114214. doi: 10.1016/j.intimp.2025.114214. Epub 2025 Feb 13.
The incidence of acute gouty arthritis (AGA) is annually increasing, significantly detrimenting the quality of life for patients. ZeXie decoction (ZXT), composed of Atractylodes macrocephala Koidz and Alisma rhizome (Sam.), a timeless formula detailed in "Synopsis of the Golden Chamber" of Chinese medical sage Zhong-Jing Zhang, has shown promising clinical application in treating AGA. Alisol B, a principal component of ZXT, remains however, elusive in its mechanism of action against AGA. This study aimed to delve into the anti-inflammatory effects of Alisol B, a key component within ZXT, and to clarify its mechanism of action in the treatment of AGA.
We adopted a network pharmacology approach to pinpoint the core targets and pathways involved in ZXT and Alisol B's treatment of AGA patients. Molecular docking was conducted using Autodock software to investigate potential interactions between Alisol B and its target proteins. An in vitro inflammation model was subsequently established. The impact of Alisol B on the expression of inflammatory factors in BMDMs treated with MSU was evaluated using RT-qPCR, supplemented by comparison with the PI3K agonist 740 Y-P (740YPDGFR) treated BMDMs. Subsequently, the expression of EGFR, PIK3CA, PIK3CB, and JAK2 - key players in the PI3K/AKT/mTOR signaling pathway - was assessed via RT-qPCR and Western blotting. Finally, the effect of MSU treatment and Alisol B's treatment on macrophage polarization was determined by flow cytometry.
Findings from network pharmacology and molecular docking suggest that Alisol B may modulate the PI3K-AKT-mTOR signaling pathway to treat AGA. In vitro experiments revealed that Alisol B inhibited the expression of inflammatory vesicles and pro-inflammatory factors by suppressing MSU-induced activation of the PI3K/AKT/mTOR signaling pathway. Additionally, Alisol B improved the cellular inflammatory environment, fostering the production of M2 cells, which could potentially repair cells within the inflammatory environment.
Our research unveils that Alisol B curtails the production of inflammatory vesicles and pro-inflammatory cytokines while enhancing the production of anti-inflammatory factors by targeting the PI3K-AKT-mTOR signaling pathway in BMDMs. This may elucidate the pivotal mechanism of Alisol B in the treatment of AGA.
急性痛风性关节炎（AGA）的发病率逐年上升，严重损害患者生活质量。泽泻汤（ZXT）由白术和泽泻组成，是中国医学圣人张仲景《金匮要略》中记载的经典方剂，在治疗AGA方面显示出良好的临床应用前景。然而，泽泻汤的主要成分泽泻醇B对AGA的作用机制尚不清楚。本研究旨在探讨泽泻汤关键成分泽泻醇B的抗炎作用，并阐明其治疗AGA的作用机制。
我们采用网络药理学方法，确定泽泻汤和泽泻醇B治疗AGA患者所涉及的核心靶点和途径。使用Autodock软件进行分子对接，研究泽泻醇B与其靶蛋白之间的潜在相互作用。随后建立体外炎症模型。使用RT-qPCR评估泽泻醇B对用MSU处理的骨髓来源巨噬细胞（BMDMs）中炎症因子表达的影响，并与用PI3K激动剂740 Y-P（740YPDGFR）处理的BMDMs进行比较。随后，通过RT-qPCR和蛋白质印迹法评估PI3K/AKT/mTOR信号通路中的关键分子表皮生长因子受体（EGFR）、磷脂酰肌醇-3激酶催化亚基α（PIK3CA）、磷脂酰肌醇-3激酶催化亚基β（PIK3CB）和Janus激酶2（JAK2）的表达。最后，通过流式细胞术确定MSU处理和泽泻醇B处理对巨噬细胞极化的影响。
网络药理学和分子对接结果表明，泽泻醇B可能通过调节PI3K-AKT-mTOR信号通路来治疗AGA。体外实验表明，泽泻醇B通过抑制MSU诱导的PI3K/AKT/mTOR信号通路激活，抑制炎症小体和促炎因子的表达。此外，泽泻醇B改善了细胞炎症环境，促进了M2细胞的产生，这可能有助于修复炎症环境中的细胞。
我们的研究表明，泽泻醇B通过靶向BMDMs中的PI3K-AKT-mTOR信号通路，减少炎症小体和促炎细胞因子的产生，同时增加抗炎因子的产生。这可能阐明了泽泻醇B治疗AGA的关键机制。
