Liu Juan, Zhang Dandan, Zhou Yaochuan, Wu Jinlu, Feng Wuwen, Peng Cheng
State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610032, China.
Chin Med. 2025 May 15;20(1):64. doi: 10.1186/s13020-025-01123-z.
Rheumatoid arthritis (RA) with cold pattern is an important type of RA according to the theory of traditional Chinese medicine. Fuzi (also known as the lateral roots of Aconitum carmichaelii Debx.) represents a typical traditional Chinese medicine that has been clinically used for treatment of the RA especially cold-related RA for thousands of years, yet its mechanism remains unknown.
The purpose of the research was to study the therapeutic effects of Fuzi on cold-related RA, and to investigate the mechanism of its action.
Here, we investigated the pharmacological effects of Fuzi on cold-related RA using micro-CT, histopathological analysis, and inflammatory cytokine test. Then, a gut microbiota composition analysis in combination with fecal microbiota transplantation were used to confirm the role of gut microbiota in the therapeutic effects of Fuzi. Further, targeted bile acid metabolomics was used to screen the possible differential microbial bile acids involved in the mechanism of Fuzi. In vitro bioactivity analysis of differential bile acids was used to assess their anti-inflammation activity. Finally, western blot was used to investigate the signaling pathways of Fuzi in reducing the inflammation of cold-related RA.
The results showed that Fuzi alleviates cold-related RA by improving arthritis index, paw swelling, bone damage, and inflammatory cytokines. In addition, the ameliorative effect of Fuzi is dependent on gut microbiota such as the taxa Lachnospiraceae and Ruminococcaceae. Targeted analysis of fecal and serum bile acids showed that TCA and THDCA were the main differential metabolites. In vitro, TCA and THDCA showed anti-inflammation effects on RAW264.7 cells. Western blot showed that Fuzi regulates TGR5-cAMP-PKA signaling and NLRP3 inflammasome to reduce cold-related arthritis.
Overall, our results demonstrated that Fuzi could regulate gut microbiota and microbial bile acid metabolism, the microbial metabolite THDCA acts on TGR5-cAMP-PKA signaling pathway and NLRP3 inflammasome to reduce cold-related arthritis. Our study suggests that supplementation of Fuzi or THDCA can be of great value for the prevention and clinical treatment of cold-related RA.
根据中医理论，寒证类风湿关节炎（RA）是RA的一种重要类型。附子（又称乌头侧根）是一种典型的中药，数千年来一直用于临床治疗RA，尤其是与寒相关的RA，但其作用机制尚不清楚。
本研究旨在探讨附子对寒证RA的治疗作用及其作用机制。
在此，我们使用微型计算机断层扫描、组织病理学分析和炎性细胞因子检测，研究附子对寒证RA的药理作用。然后，结合粪便微生物群移植进行肠道微生物群组成分析，以确认肠道微生物群在附子治疗作用中的作用。此外，使用靶向胆汁酸代谢组学筛选附子作用机制中可能存在差异的微生物胆汁酸。对差异胆汁酸进行体外生物活性分析，以评估其抗炎活性。最后，采用蛋白质免疫印迹法研究附子减轻寒证RA炎症的信号通路。
结果表明，附子通过改善关节炎指数、足爪肿胀、骨损伤和炎性细胞因子来减轻寒证RA。此外，附子的改善作用依赖于肠道微生物群，如毛螺菌科和瘤胃球菌科的分类群。对粪便和血清胆汁酸的靶向分析表明，三羟基胆酸（TCA）和四羟基脱氧胆酸（THDCA）是主要的差异代谢产物。在体外，TCA和THDCA对RAW264.7细胞具有抗炎作用。蛋白质免疫印迹法显示，附子通过调节TGR5-cAMP-PKA信号通路和NLRP3炎性小体来减轻寒证关节炎。
总体而言，我们的结果表明，附子可以调节肠道微生物群和微生物胆汁酸代谢，微生物代谢产物THDCA作用于TGR5-cAMP-PKA信号通路和NLRP3炎性小体，以减轻寒证关节炎。我们的研究表明，补充附子或THDCA对寒证RA的预防和临床治疗具有重要价值。
