南方医科大学学报

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (10): 2434-2446.doi: 10.12122/j.issn.1673-4254.2026.10.13

• • 上一篇    

加味清络饮通过调控Hedgehog信号通路促进滑膜细胞凋亡缓解大鼠类风湿性关节炎病变

郝易飞1(), 蔡炼清2, 唐丝雨2, 于晓萱2, 李艳1, 王秀1(), 朱洁1,2,3()   

  1. 1.皖南医科大学中西医结合研究中心，安徽 芜湖 241000
    2.安徽中医药大学中西医结合学院，安徽 合肥 230000
    3.安徽中医药大学新安医学教育部重点实验室，安徽 合肥 230038
  • 收稿日期:2026-03-26 出版日期:2026-10-20 发布日期:2026-10-26
  • 通讯作者: 王秀,朱洁 E-mail:3462857600@qq.com;wangxiuu@sina.com;janezhutcm@foxmail.com
  • 作者简介:郝易飞，在读硕士研究生，E-mail: 3462857600@qq.com
  • 基金资助:
    国家自然科学基金(82274465);安徽省中医药科技攻关专项项目(202303a07020001);安徽省高校杰出青年科研项目(2022AH020044)

Jiawei Qingluo Yin alleviates rheumatoid arthritis in rats by inhibiting activation of the Hedgehog signaling pathway to promote apoptosis of synovial cells

Yifei HAO1(), Lianqing CAI2, Siyu TANG2, Xiaoxuan YU2, Yan LI1, Xiu WANG1(), Jie ZHU1,2,3()   

  1. 1.Research Center of Integrated Traditional Chinese and Western Medicine, Wannan Medical University, Wuhu 241000, China
    2.School of Integrated Traditional Chinese and Western Medicine, Anhui University of Chinese Medicine, Hefei 230000, China
    3.Key Laboratory of Xin'an Medicine, Anhui University of Chinese Medicine, Hefei 230038, China
  • Received:2026-03-26 Online:2026-10-20 Published:2026-10-26
  • Contact: Xiu WANG, Jie ZHU E-mail:3462857600@qq.com;wangxiuu@sina.com;janezhutcm@foxmail.com
  • Supported by:
    National Natural Science Foundation of China(82274465)

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摘要：

目的 基于网络药理学、分子对接、分子动力学模拟与动物实验相结合的多技术融合策略探究加味清络饮（JWQLY）调控Hedgehog信号通路促进滑膜细胞凋亡治疗类风湿性关节炎（RA）的作用机制研究。 方法 通过网络药理学分析确定JWQLY治疗RA的核心成分、关键靶点及信号通路，采用分子对接和分子动力学模拟对核心成分与关键靶点的结合活性进行验证。采用完全弗氏佐剂诱导建立AIA大鼠模型，在疗效观察阶段，将造模成功的大鼠随机分为正常组、模型组、甲氨蝶呤（MTX）（0.5 mg·kg-1·d-1）组、JWQLY低、中、高剂量（1.03、3.1、9.3 g·kg-1·d-1）组。观察大鼠一般生理指标、足爪肿胀度及关节炎指数（AI）评分；采用HE染色观察滑膜组织病理学变化；采用TUNEL法检测细胞凋亡；采用ELISA法检测血清白细胞介素-1β（IL-1β）、血管内皮生长因子（VEGF）水平。在机制验证阶段，将造模成功的AIA大鼠模型随机分为正常组、模型组、JWQLY高（9.3 g·kg-1·d-1）组、抑制剂组（维莫德吉，10 mg·kg-1·d-1）及JWQLY+抑制剂组。观察大鼠一般生理指标、足爪肿胀度及关节炎指数（AI）评分；采用HE染色观察滑膜组织病理学变化；采用免疫组化法检测滑膜组织Bcl-2、Bax蛋白表达；采用Western blotting法检测滑膜组织Ihh、Smo、Gli1、cleaved-caspase-3、cleaved-caspase-9蛋白表达。 结果 通过网络药理学筛选JWQLY靶点518个、RA疾病靶点4210个，交集靶点306个。GO富集分析显示靶点主要涉及凋亡过程的负调控、炎症反应等生物过程；KEGG通路富集分析显示靶点显著富集于Hedgehog信号通路以及凋亡信号通路。分子对接及分子动力学模拟结果表明，核心成分与Hedgehog通路关键靶点Shh等具有稳定的结合活性。疗效观察阶段结果显示，与正常组比较，模型组大鼠足爪肿胀明显，AI评分、病理评分、血清IL-1β及VEGF水平显著升高（P<0.05），且呈剂量依赖性。机制验证阶段结果显示，与模型组比较，抑制剂组和JWQLY+抑制剂组滑膜组织Ihh、Smo、Gli1、Bcl-2蛋白表达显著降低（P<0.05），Bax、cleaved-caspase-3、cleaved-caspase-9蛋白表达及细胞凋亡率显著升高（P<0.05）；JWQLY与抑制剂联用后对Hedgehog通路产生协同抑制作用，进一步促进滑膜细胞凋亡。 结论 JWQLY通过抑制Hedgehog信号通路激活，下调Bcl-2/Bax比值，促进滑膜细胞凋亡，进而缓解类风湿关节炎病变。

关键词: 加味清络饮, 类风湿性关节炎, Hedgehog信号通路, 滑膜细胞, 凋亡

Abstract:

Objective To investigate the mechanism of Jiawei Qingluo Yin (JWQLY) for alleviating rheumatoid arthritis (RA) in rats. Methods Network pharmacology, molecular docking and dynamics analyses were performed to identify the core components, key targets, and pathways of JWQLY against RA and the core component-key target binding. In a rat model of antigen-induced arthritis (AIA) established using complete Freund's adjuvant, the effects of MTX (0.5 mg/kg) and low-, medium-, and high-dose JWQLY (1.03, 3.1, and 9.3 g/kg, respectively) were tested on paw swelling, arthritis index (AI), synovial histopathology, apoptosis, and serum IL-1β and VEGF levels. In the mechanistic investigation, the rat models were treated with high-dose (9.3 g/kg) JWQLY, vismodegib (a Hedgehog inhibitor, 10 mg/kg), or both, and the changes in paw swelling, AI, histopathology, and expressions of Bcl-2, Bax, Ihh, Smo, Gli1, cleaved caspase-3, and cleaved caspase-9 were examined. Results A total of 306 intersecting targets were identified between JWQLY and RA. GO and KEGG enrichment analyses revealed enrichment of the targets in negative regulation of apoptosis and inflammation and in Hedgehog and apoptosis pathways. Molecular docking studies confirmed stable binding of the core components in JWQLY to Shh and other Hedgehog targets. The rat models of AIA showed obvious paw swelling and significantly elevated AI, pathology scores, and serum IL-1β and VEGF levels, and these changes were dose-dependently alleviated by JWQLY treatment. The rat models receiving treatment with vismodegib or JWQLY+ vismodegib exhibited significantly reduced Ihh, Smo, Gli1, and Bcl-2 expression with increased expressions of Bax, cleaved- caspase-3, and cleaved-caspase-9 and apoptosis rates of the synovial cells. JWQLY and vismodegib synergistically suppressed Hedgehog signaling and enhanced synovial cell apoptosis. Conclusion JWQLY alleviates RA in rats by inhibiting Hedgehog pathway activation, down-regulating the Bcl-2/Bax ratio, and promoting synovial cell apoptosis.

Key words: Jiawei Qingluo Yin, rheumatoid arthritis, Hedgehog signaling pathway, synovial cells, apoptosis