网络药理学与分子对接联合实验验证的加味二妙散抗急性痛风性关节炎机制研究

doi:10.12092/j.issn.1009-2501.2026.03.003

中国临床药理学与治疗学 ›› 2026, Vol. 31 ›› Issue (3): 313-323.doi: 10.12092/j.issn.1009-2501.2026.03.003

网络药理学与分子对接联合实验验证的加味二妙散抗急性痛风性关节炎机制研究

刘晓绵¹(), 刘颖¹, 涂文婧¹, 张硕¹, 朱高彦¹, 崔语嫣¹, 崔肖雨¹, 杨一璇¹, 李晓冰²^,*(), 郭洪涛³, 李栋⁴, 何小鹃⁵

1. 河南中医药大学医学院，郑州 450046，河南
2. 河南中医药大学中医学院，郑州 450046，河南
3. 河南中医药大学第一附属医院风湿病科，郑州 450000，河南
4. 河南中医药大学第一临床医学院，郑州 450046，河南
5. 中国中医科学院中医临床基础医学研究所，北京 100700

收稿日期:2025-05-09 修回日期:2025-08-28 出版日期:2026-03-26 发布日期:2026-04-03
通讯作者: 李晓冰 E-mail:lxmianjyjy@163.com;baishaoyao@163.com
作者简介:刘晓绵，女，研究方向：中西医结合防治风湿免疫病。E-mail：lxmianjyjy@163.com
基金资助:
国家自然科学基金面上项目（82274342）；河南省中医药传承与创新人才工程（仲景工程）中医药学科拔尖人才项目（CZ0325-13）；河南省中医药科学研究专项项目（2023ZXZX1145）；河南省科技攻关项目（252102311259）；河南省大学生创新训练项目（202410471043）；河南中医药大学科研苗圃（MP2024-49）；河南中医药大学科研苗圃（MP2023-34）

Unraveling the molecular mechanisms of modified Er-Miao-San (MEMS) in acute gouty arthritis: a multidisciplinary integration of network pharmacology, molecular docking, and experimental validation

Xiaomian LIU¹(), Ying LIU¹, Wenjing TU¹, Shuo ZHANG¹, Gaoyan ZHU¹, Yuyan CUI¹, Xiaoyu CUI¹, Yixuan YANG¹, Xiaobing LI²^,*(), Hongtao GUO³, Dong LI⁴, Xiaojuan HE⁵

1. School of Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, Henan, China
2. School of Traditional Chinese Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, Henan, China
3. Department of Rheumatology, The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000, Henan, China
4. School of Clinical Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, Henan, China
5. Institute of Clinical Basic Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China

Received:2025-05-09 Revised:2025-08-28 Online:2026-03-26 Published:2026-04-03
Contact: Xiaobing LI E-mail:lxmianjyjy@163.com;baishaoyao@163.com

摘要/Abstract

摘要：

目的: 探讨加味二妙散（modified Er Miao-San，MEMS）治疗急性痛风性关节炎（acute gouty arthritis，AGA）的疗效及潜在作用机制。方法: 通过网络药理学、分子对接和分子动力学模拟验证MEMS活性成分与AGA靶点间的相互作用。通过向大鼠右踝关节注射25 mg/kg尿酸钠（MSU）悬液建立AGA模型。通过测量踝关节肿胀度、步态评分和炎症指数评估治疗效果；ELISA法测定AGA大鼠血清TNF-α、IL-6、IL-1β、IL-17、MDA、SOD和GSH浓度；HE染色观察踝关节滑膜病理变化；Western blot分析各组踝关节滑膜NLRP3和STAT3蛋白水平。结果: 网络药理学分析发现，槲皮素、β-谷甾醇、豆甾醇和汉黄芩素是MEMS治疗AGA的核心活性成分。TNF、STAT3、IL-6和IL-1β是MEMS治疗AGA的关键靶点，核心活性成分与关键靶点具有较强的结合活性和稳定构象。关键靶点主要富集于NOD样受体、IL-17和TNF等信号通路。动物模型研究表明，MEMS能有效减轻AGA大鼠踝关节肿胀度，改善步态评分和炎症指数，减轻踝关节滑膜细胞增殖、中性粒细胞浸润和毛细血管充血，降低血清TNF-α、IL-1β、IL-6、IL-17和MDA水平，提高SOD和GSH含量，下调踝关节滑膜NLRP3和STAT3蛋白表达。结论: MEMS能减轻AGA大鼠炎症和氧化应激损伤，发挥抗炎抗氧化功能，其作用机制可能与通过抑制STAT3/NLRP3信号通路调控炎症因子水平有关。

关键词: 急性痛风性关节炎, 加味二妙散, 网络药理学, 分子对接, 分子动力学

Abstract:

AIM: To investigate the therapeutic efficacy and potential mechanisms of modified Er Miao-San (MEMS) in the treatment of Acute Gouty Arthritis (AGA). METHODS: Network pharmacology, molecular docking, and molecular dynamics simulations were used to validate the interactions between the active components of MEMS and the targets associated with AGA. An AGA model was established in rats by injecting 25 mg/kg of monosodium urate (MSU) suspension into the right ankle joint. Treatment efficacy was evaluated by measuring ankle joint swelling, gait scores, and inflammation indices. Enzyme-linked immunosorbent assay (ELISA) was used to determine the serum levels of TNF-α, IL-6, IL-1β, IL-17, MDA, SOD, and GSH in AGA rats. Hematoxylin and eosin (HE) staining was performed to observe the pathological changes in the ankle joint synovial tissue, and Western blotting (WB) was used to analyze the protein levels of NLRP3 and STAT3 in the synovial tissue. RESULTS: Network pharmacology analysis identified quercetin, β-sitosterol, stigmasterol, and wogonin as the core active components in MEMS for the treatment of AGA. The key therapeutic targets of MEMS in treating AGA include TNF, STAT3, IL-6, and IL-1β. The core active components exhibited strong binding affinity and stable conformations with these targets. The key targets were predominantly enriched in signaling pathways, including NOD-like receptors, IL-17, and TNF. In vivo experiments demonstrated that MEMS effectively reduced ankle joint swelling, improved gait scores, alleviated synovial cell proliferation, neutrophil infiltration, and capillary congestion in AGA rats. It also lowered the serum levels of TNF-α, IL-1β, IL-6, IL-17, and MDA, while increasing the levels of SOD and GSH. Furthermore, MEMS downregulated the expression of NLRP3 and STAT3 proteins in the synovial tissue. CONCLUSION: MEMS alleviates inflammation and oxidative stress in AGA rats, exerting anti-inflammatory and antioxidative effects. Its mechanism of action may be associated with the modulation of inflammation factor levels via the inhibition of the STAT3/NLRP3 signaling pathway.

Key words: acute gouty arthritis, modified Er Miao-San, network pharmacology, molecular docking, molecular dynamics

中图分类号:

R965.2

刘晓绵, 刘颖, 涂文婧, 张硕, 朱高彦, 崔语嫣, 崔肖雨, 杨一璇, 李晓冰, 郭洪涛, 李栋, 何小鹃. 网络药理学与分子对接联合实验验证的加味二妙散抗急性痛风性关节炎机制研究[J]. 中国临床药理学与治疗学, 2026, 31(3): 313-323.

Xiaomian LIU, Ying LIU, Wenjing TU, Shuo ZHANG, Gaoyan ZHU, Yuyan CUI, Xiaoyu CUI, Yixuan YANG, Xiaobing LI, Hongtao GUO, Dong LI, Xiaojuan HE. Unraveling the molecular mechanisms of modified Er-Miao-San (MEMS) in acute gouty arthritis: a multidisciplinary integration of network pharmacology, molecular docking, and experimental validation[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(3): 313-323.

图/表 15

图 1

Fig.1 Venny diagram of the constituent target of MEMS and AGA target

图 2

Fig.2 Protein-protein interaction network

图 3

Fig.3 The TCM-active components-drug targets network Rectangles represent the intersection targets of drugs and diseases, triangles represent the core active components of drugs, rhombuses represent the categories of traditional Chinese medicine, and concave quadrilaterals represent the common active components of drugs.

表 1

Table 1 Core active compounds of MEMS and their degree values

Number	Molecule ID	Molecule name	OB(%)	DL	TCM	Degree
1	MOL000098	Quercetin	46.43	0.28	HuangBai, TuFuLing	51
2	MOL000358	beta-sitosterol	36.91	0.75	HuangBai, TuFuLing, ShanCiGu	37
3	MOL000173	wogonin	30.68	0.23	CangZhu	29
4	MOL000449	Stigmasterol	43.83	0.76	HuangBai, TuFuLing, ShanCiGu	29

图 4

Fig.4 GO enrichment analysis bar diagram

图 5

Fig.5 KEGG enrichment analysis diagram

图 6

Fig.6 Molecular docking binding energy heat map (kcal/mol)

图 7

Fig.7 molecular docking diagram A,B: docking diagram of beta-sitosterol and target TNF; C,D: docking diagram of stigmasterol and target TNF.

图 8

Fig.8 Conformational stability analysis of TNF in complex with β-sitosterol or stigmasterol A-C: RMSD, RMSF, and Rg profiles of the TNF-β-sitosterol complex; D-F: corresponding profiles of the TNF-stigmasterol complex.

图 9

Fig.9 Hydrogen bonding and binding free energy analysis A,B: number of hydrogen bonds in the TNF-β-sitosterol and TNF-stigmasterol complexes, respectively. C,D: binding free energy decomposition. E,F: per-residue energy contributions for each complex.

图 10

Fig.10 The free energy morphology of TNF-β-sitosterol and TNF-stigmasterol complexes A: TNF-β-sitosterol; B: TNF-stigmasterol.

图 11

Fig.11 Effects of MEMS on ankle joint swelling, toe volume swelling, gait score, and inflammatory index in AGA rats A: ankle swelling before and 12 h after MSU injection; B: comparison of the degree of right posterior ankle joint swelling, the degree of toe volume ankle swelling, gait assessment score and Inflammatory index in rats of different group ($ \overline{x} $±s, n=6). bP<0.05, cP<0.01, compared with model group; eP<0.05, compared with EMS group.

图 12

Fig.12 Results of various indexes in rats ($ \overline{x} $±s, n=6) A: TNF-α; B: IL-1β; C: IL-6; D: IL-17; E: SOD; F: GSH; G: MDA. bP<0.05, cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with Model group; hP<0.05, compared with EMS group.

图 13

Fig.13 HE staining of synovial tissues in rat ankle joints across experimental groups ($ \overline{x} $±s, n=6) The representative image of HE staining in synovial tissue (Scale bar=100 μm); the histological quantitative score of synovial tissue sections was average±SD. cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with model group.

图 14

Fig.14 The expression levels of NLRP3 and STAT3 protein in ankle joint tissue of rats ($ \overline{x} $±s, n=6) A: STAT3 protein expression; B: NLRP3 protein expression; C: electrophoresis of STAT3 and NLRP3 proteins. cP<0.01, compared with control group; fP<0.01, compared with model group; iP<0.01, compared with EMS group.

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网络药理学与分子对接联合实验验证的加味二妙散抗急性痛风性关节炎机制研究

Unraveling the molecular mechanisms of modified Er-Miao-San (MEMS) in acute gouty arthritis: a multidisciplinary integration of network pharmacology, molecular docking, and experimental validation

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