中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (25): 6632-6642.doi: 10.12307/2026.287

• 组织工程相关大数据分析 Big data analysis in tissue engineering • 上一篇    下一篇

补肾强筋胶囊调控炎症信号通路改善膝骨关节炎的作用机制

何汶洋1，2，陈伟坚2，冉清智3，黄译蝶1，2，林晓东2，许学猛2，郭美容2，刘文刚1，2   

  1. 1广州中医药大学，广东省广州市   510405；2广东省第二中医院(广东省中医药工程技术研究院)，广东省广州市   510095；3中国中医科学院广安门医院，北京市   100053 
  • 收稿日期:2025-10-15 修回日期:2025-12-26 出版日期:2026-09-08 发布日期:2026-04-23
  • 通讯作者: 刘文刚，主任医师，博士生导师，广州中医药大学，广东省广州市 510405；广东省第二中医院(广东省中医药工程技术研究院)，广东省广州市 510095
  • 作者简介:何汶洋，女，2000年生，四川省成都市人，汉族，广州中医药大学在读硕士，主要从事中医骨伤科学研究。
  • 基金资助:
    广东省基础与应用基础研究基金-自然科学基金项目面上项目(2023A1515012615)，项目负责人：刘文刚；广东省基础与应用基础研究基金-省企联合基金-面上项目(2022A1515220157)，项目负责人：刘文刚；广东省第二中医院科研创新基金-卓越团队项目(SEZYY2023B16)，项目负责人：刘文刚；广东省医学科学技术研究基金项目(B2023352)，项目负责人：林晓东；广东省中医药局课题(20221033)，项目负责人：郭美容

Action mechanisms of Bushen Qiangjin capsule in regulating inflammatory signaling pathways and improving knee osteoarthritis

He Wenyang1, 2, Chen Weijian2, Ran Qingzhi3, Huang Yidie1, 2, Lin Xiaodong2, Xu Xuemeng2, Guo Meirong2, Liu Wengang1, 2   

  1. 1Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 2Guangdong Second Traditional Chinese Medicine Hospital (Guangdong Province Engineering Technology Research Institute of TCM), Guangzhou 510095, Guangdong Province, China; 3Guang'anmen Hospital of China Academy of Chinese Medical Sciences, Beijing 100053, China

  • Received:2025-10-15 Revised:2025-12-26 Online:2026-09-08 Published:2026-04-23
  • Contact: Liu Wengang, Chief physician, Doctoral supervisor, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; Guangdong Second Traditional Chinese Medicine Hospital (Guangdong Province Engineering Technology Research Institute of TCM), Guangzhou 510095, Guangdong Province, China
  • About author:He Wenyang, MS candidate, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; Guangdong Second Traditional Chinese Medicine Hospital (Guangdong Province Engineering Technology Research Institute of TCM), Guangzhou 510095, Guangdong Province, China
  • Supported by:

    Guangdong Provincial Basic and Applied Basic Research Foundation - Natural Science Foundation Project, No. 2023A1515012615 (to LWG); Guangdong Provincial Basic and Applied Basic Research Foundation - Provincial Enterprise Joint Fund - General Project, No. 2022A1515220157 (to LWG); Guangdong Provincial Second Affiliated Hospital of Traditional Chinese Medicine Scientific Research Innovation Fund - Excellent Team Project, No. SEZYY2023B16 (to LWG); Guangdong Provincial Medical Science and Technology Research Foundation Project, No. B2023352 (to LXD); Guangdong Provincial Administration of Traditional Chinese Medicine Project, No. 20221033 (to GMR) 

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摘要：



文题释义：
膝骨关节炎：以关节软骨退变、关节边缘骨赘形成及滑膜病变为主要病理特征的慢性退行性疾病。
基于汇总数据的孟德尔随机化分析：是一种整合基因表达数量性状与表型全基因组关联分析数据的方法，用于评估基因表达与疾病或性状之间的潜在因果关系。

背景：动物实验证明，补肾强筋胶囊可显著改善软骨下骨代谢及异常骨重塑、延缓膝骨关节炎的进展，为阐释该药治疗膝骨关节炎的机制提供了实验依据。
目的：通过网络药理学、基于汇总数据的孟德尔随机化、两样本孟德尔随机化、分子对接及细胞实验验证等方法，系统探讨补肾强筋胶囊调控炎症信号通路改善膝骨关节炎的作用机制。
方法：①通过网络药理学工具获得补肾强筋胶囊治疗膝骨关节炎的有效成分、核心靶点和信号通路，将潜在作用靶点导入STRING平台构建蛋白互作网络，进一步筛选核心靶点。开展基于汇总数据的孟德尔随机化分析，评估核心靶点与膝骨关节炎发病风险的因果关系；采用两样本孟德尔随机化分析丝裂原活化蛋白激酶8与膝骨关节炎的因果关系。对补肾强筋胶囊干预膝骨关节炎的潜在靶标进行GO功能和KEGG通路富集分析。利用分子对接进一步明确补肾强筋胶囊关键活性成分与核心靶标蛋白之间的相互作用。②取5只SD大鼠，灌胃给予胃补肾强筋胶囊0.243 g/(kg·d)，连续给药7 d。末次给药结束后采集静脉血，分离血清获得含药血清。将大鼠膝关节软骨细胞分3组培养：空白组不进行任何处理，模型组加入白细胞介素1β处理24 h建立骨关节炎细胞模型，含药血清组加入白细胞介素1β处理24 h后加入体积分数10%含药血清处理24 h。处理结束后，阿利新蓝和甲苯胺蓝细胞染色观察软骨细胞形态，RT-qPCR、Western blot检测丝裂原活化蛋白激酶8、Ⅱ型胶原的mRNA与蛋白表达。
结果与结论：①共获得补肾强筋胶囊相关的活性成分85个，其中山柰酚、阿魏酸为关键成分，核心作用靶点包括丝氨酸/苏氨酸蛋白激酶1、磷脂酰肌醇3–激酶催化亚基α等。基于汇总数据的孟德尔随机化、两样本孟德尔随机化分析提示，丝裂原活化蛋白激酶8与膝骨关节炎存在直接因果关系，并且呈正相关。GO富集结果主要涉及磷酸化、对异生物刺激反应、细胞凋亡过程负调控等生物过程，KEGG通路主要包括磷脂酰肌醇3–激酶–丝氨酸/苏氨酸蛋白激酶、肿瘤坏死因子α、Toll样受体4、缺氧诱导因子1及机械感受相关轴等信号通路。分子对接结果显示，山柰酚、阿魏酸与丝氨酸/苏氨酸蛋白激酶1、磷脂酰肌醇3–激酶催化亚基α、酪氨酸激酶C、信号转导及转录激活因子3、肿瘤蛋白p53等核心靶点的结合活性良好。②阿利新蓝和甲苯胺蓝细胞染色显示，补肾强筋胶囊可促进骨关节炎细胞模型软骨外基质的合成。RT-qPCR、Western blot检测显示，相较于模型组，含药血清组细胞中Ⅱ型胶原的mRNA与蛋白表达升高(P < 0.05)，丝裂原活化蛋白激酶8的mRNA与蛋白表达降低(P < 0.05)。③结果表明，补肾强筋胶囊可能通过“多成分–多靶点–多通路”协同调控炎症反应与细胞外基质稳态，从而改善膝骨关节炎相关损伤。
https://orcid.org/0009-0003-7322-3643(何汶洋)；https://orcid.org/0000-0002-8887-8429(刘文刚)


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 膝骨关节炎, 补肾强筋胶囊, 网络药理学, 孟德尔随机化, 分子对接, 细胞实验

Abstract: BACKGROUND: Animal experiments have demonstrated that Bushen Qiangjin capsule can significantly improve subchondral bone metabolism and abnormal bone remodeling, and delay the progression of knee osteoarthritis, providing experimental evidence for elucidating the mechanism of action of this drug in treating knee osteoarthritis.
OBJECTIVE: To systematically explore the mechanism of action of Bushen Qiangjin capsule in regulating inflammatory signaling pathways to improve knee osteoarthritis using network pharmacology, Mendelian randomization based on pooled data, two-sample Mendelian randomization, molecular docking, and cell experiments.
METHODS: (1) The effective components, core targets, and signaling pathways of Bushen Qiangjin capsule in treating knee osteoarthritis were obtained using network pharmacology tools. Potential targets were imported into the STRING platform to construct a protein interaction network for further screening of core targets. Mendelian randomization analysis based on pooled data was conducted to assess the causal relationship between core targets and the risk of knee osteoarthritis. Two-sample Mendelian randomization was used to analyze the causal relationship between mitogen-activated protein kinase 8 and knee osteoarthritis. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on potential targets of Bushen Qiangjin capsule in intervening in knee osteoarthritis. Molecular docking was used to further clarify the interaction between the key active ingredients of Bushen Qiangjin capsule and its core target protein. (2) Five Sprague-Dawley rats were administered Bushen Qiangjin capsule at a dose of 0.243 g/kg per day via gavage for 7 consecutive days. Venous blood was collected after the last administration, and serum containing the drug was separated. Chondrocytes from the rat knee were cultured in three groups: a blank group received no treatment; a model group was treated with interleukin-1β for 24 hours to establish an osteoarthritis cell model; and a serum-containing group was treated with interleukin-1β for 24 hours followed by treatment with 10% drug-containing serum (volume fraction) for 24 hours. After treatment, chondrocyte morphology was observed by alicin blue and toluidine blue staining. The mRNA and protein expression of mitogen-activated protein kinase 8 and type II collagen were detected by RT-qPCR and western blot assay.
RESULTS AND CONCLUSION: (1) A total of 85 active ingredients related to Bushen Qiangjin capsule were obtained, among which kaempferol and ferulic acid were key components. Core targets included serine/threonine protein kinase 1 and phosphatidylinositol 3-kinase catalytic subunit α. Mendelian randomization and two-sample Mendelian randomization analyses based on the aggregated data indicated a direct causal relationship between mitogen-activated protein kinase 8 and knee osteoarthritis, and this relationship was positive. Gene Ontology enrichment results mainly involved biological processes such as phosphorylation, response to heterobiotic stimuli, and negative regulation of apoptosis. The Kyoto Encyclopedia of Genes and Genomes pathways mainly included signaling pathways such as phosphatidylinositol 3-kinase-serine/threonine protein kinase, tumor necrosis factor α, Toll-like receptor 4, hypoxia-inducible factor 1, and mechanosensory-related axes. Molecular docking results showed that kaempferol and ferulic acid exhibited good binding activity to core targets such as serine/threonine protein kinase 1, phosphatidylinositol 3-kinase catalytic subunit α, tyrosine kinase C, signal transduction and transcription activator 3, and tumor protein p53. (2) Alcian blue and toluidine blue staining showed that Bushen Qiangjin capsule promoted the synthesis of extracellular matrix in an osteoarthritis cell model. RT-qPCR and western blot analysis showed that, compared with the model group, the mRNA and protein expression levels of type II collagen were increased in the serum-containing group (P < 0.05), while the mRNA and protein expression levels of mitogen-activated protein kinase 8 were decreased (P < 0.05). (3) The results indicate that Bushen Qiangjin capsule may improve knee osteoarthritis-related damage by synergistically regulating inflammatory responses and extracellular matrix homeostasis through a multi-component, multi-target, and multi-pathway approach.

Key words: knee osteoarthritis, Bushen Qiangjin capsule, network pharmacology, Mendelian randomization, molecular docking, cell experiment

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