中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (34): 8868-8877.doi: 10.12307/2026.893

• 骨组织构建 bone tissue construction • 上一篇    下一篇

木犀草素调控巨噬细胞极化治疗膝骨关节炎的机制

赵灿斌，曾  平，石炜琦，刘金富，丁  强，郭  良，王伟伟，陶红成，郭亚沨，秦  英   

  1. 1广西中医药大学第一临床医学院，广西壮族自治区南宁市  530200；2广西中医药大学第一附属医院，广西壮族自治区南宁市  530200；3暨南大学中医学院，广东省广州市  510623；4济南市长清区中医医院骨科，山东省济南市  250300
  • 收稿日期:2025-09-24 修回日期:2026-02-14 出版日期:2026-12-08 发布日期:2026-04-11
  • 通讯作者: 秦英，硕士，主治医师，济南市长清区中医医院骨科，山东省济南市 250300
  • 作者简介:赵灿斌，男，1997年生，山东省泰安市人，汉族，广西中医药大学2024级博士在读，主要从事骨关节退变与缺血性疾病的中医防治研究。
  • 基金资助:
    国家自然科学基金项目(82160913)，项目负责人：曾平；山东省自然科学基金项目(ZR2022MH147)，项目参与人：
    赵灿斌；广西自然科学基金项目(2025GXNSFBA069262)，项目参与人：王伟伟、丁强

Mechanism by which luteolin regulates macrophage polarization in the treatment of knee osteoarthritis

Zhao Canbin1, Zeng Ping2, Shi Weiqi1, Liu Jinfu3, Ding Qiang1, Guo Liang1, Wang Weiwei1, Tao Hongcheng1, Guo Yafeng1, Qin Ying4   

  1. 1First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; 2First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; 3College of Traditional Chinese Medicine, Jinan University, Guangzhou 510623, Guangdong Province, China; 4Department of Orthopedics, Jinan Changqing District Traditional Chinese Medicine Hospital, Jinan 250300, Shandong Province, China
  • Received:2025-09-24 Revised:2026-02-14 Online:2026-12-08 Published:2026-04-11
  • Contact: Qin Ying, MS, Attending physician, Department of Orthopedics, Jinan Changqing District Traditional Chinese Medicine Hospital, Jinan 250300, Shandong Province, China
  • About author:Zhao Canbin, PhD candidate, First Clinical Medical College, Guangxi University of Traditional Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 82160913 (to ZP); Natural Science Foundation of Shandong Province, No. ZR2022MH147 (to ZCB [project participant]); Natural Science Foundation of Guangxi Zhuang Autonomous Region, No. 2025GXNSFBA069262 (to WWW and DQ [project participants]) 

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摘要：



文题释义：
膝骨关节炎：是一种由于多种因素(如年龄增长、肥胖、创伤等)导致的膝关节退行性疾病。膝骨关节炎的发病机制尚不完全明确，可能与软骨细胞凋亡、炎症反应、基质代谢失衡、力学负荷异常等有关，具体的发生发展机制还有待进一步探索。
巨噬细胞极化：是巨噬细胞在微环境刺激下呈现的功能表型转换过程，主要表现为向促炎的M1型和抗炎的M2型分化。巨噬细胞极化的调控机制涉及多种细胞因子、信号通路及代谢重编程，在免疫应答、组织修复、肿瘤发展等多种生理和病理过程中发挥关键作用，包括感染清除、创伤愈合、慢性炎症性疾病等。

背景：巨噬细胞极化状态与膝骨关节炎的发生发展密切相关。木犀草素可以调控核因子κB信号通路影响巨噬细胞极化过程，但具体机制尚未明确。
目的：研究木犀草素治疗膝骨关节炎的机制。
方法：①通过网络药理学检索木犀草素、巨噬细胞极化及膝骨关节炎的交集靶点，构建蛋白质相互作用网络，并进行GO、KEGG富集分析及分子对接处理。②敲除核因子κB p65后，诱导RAW264.7细胞分别向M1与M2型极化，流式细胞学检测细胞极化倾向。采用不同浓度的木犀草素干预RAW264.7细胞，通过CCK-8法筛选适宜浓度的木犀草素进行后续实验。诱导RAW264.7细胞分别向M1与M2型极化，同时加入木犀草素干预，流式细胞学检测细胞极化倾向。诱导RAW264.7细胞向M1型极化，同时加入木犀草素干预，Elisa法检测细胞上清中肿瘤坏死因子α、白细胞介素6、白细胞介素10水平，免疫荧光染色检测核因子κB p65/p50核转位情况，Western blot检测核因子κB p65、核因子κB抑制蛋白α表达及磷酸化情况。
结果与结论：①共筛选出交集基因137个，其中135个靶基因均参与蛋白质相互作用网络构建，GO、KEGG分析显示核因子κB复合体在蛋白质相互作用网络中富集。分子对接显示，木犀草素与核因子κB抑制蛋白α、核因子κB p50/p65蛋白对接良好。②核因子κB p65敲除后，RAW264.7细胞向M1型极化倾向受到抑制，出现自发性M2型极化倾向。木犀草素干预可抑制RAW264.7细胞向M1型极化、促进RAW264.7细胞向M2型极化。诱导RAW264.7细胞向M1型极化过程中，木犀草素干预可抑制促炎因子释放、促进抗炎因子释放，抑制核因子κB p65/p50核转位，促进核因子κB抑制蛋白α表达并抑制其磷酸化，抑制核因子κB p65表达及其磷酸化。结果表明，木犀草素可通过核因子κB信号通路调控巨噬细胞极化，对膝骨关节炎有着潜在的治疗作用。
https://orcid.org/0009-0008-0004-7999(赵灿斌)


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: 木犀草素, 巨噬细胞, 核因子κB信号通路, 网络药理学, 膝骨关节炎, 巨噬细胞极化

Abstract: BACKGROUND: The polarization state of macrophages is closely related to the occurrence and development of knee osteoarthritis. Luteolin can regulate the nuclear transcription factor κB (NF-κB) signaling pathway to affect the polarization process of macrophages, but the specific mechanism remains unclear. 
OBJECTIVE: To investigate the mechanism of luteolin in the treatment of knee osteoarthritis. 
METHODS: (1) Intersectional targets of luteolin, macrophage polarization, and knee osteoarthritis were retrieved via network pharmacology. A protein-protein interaction network was constructed, followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses, and molecular docking. (2) After knocking out nuclear factor κB-light-chain-enhancer of activated B cells p65 subunit (NF-κB p65), RAW264.7 cells were induced to polarize toward M1 and M2 phenotypes, and polarization trends were detected by flow cytometry. Different concentrations of luteolin were applied to RAW264.7 cells, and the suitable concentration for subsequent experiments was selected using the cell counting kit-8 assay. RAW264.7 cells were induced to polarize toward M1 and M2 phenotypes with simultaneous luteolin intervention, and polarization trends were detected by flow cytometry. RAW264.7 cells were induced toward M1 polarization with simultaneous luteolin intervention. The levels of tumor necrosis factor-α, interleukin-6, and interleukin-10 in the cell supernatant were measured by ELISA. Nuclear translocation of NF-κB p65/p50 was detected by immunofluorescence staining. The expression and phosphorylation of NF-κB p65 and NF-κB inhibitor α (IκBα) were detected by western blot.
RESULTS AND CONCLUSION: (1) A total of 137 intersection genes were screened, among which 135 target genes participated in the construction of the protein-protein interaction network. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that the NF-κB complex was enriched in the protein-protein interaction network. Molecular docking indicated that luteolin bound well to IκBα, NF-κB p50, and NF-κB p65 proteins. (2) After NF-κB p65 knockout, the tendency of RAW264.7 cells to polarize toward the M1 phenotype was inhibited, and spontaneous polarization toward the M2 phenotype occurred. Luteolin intervention inhibited RAW264.7 cell polarization toward the M1 phenotype and promoted polarization toward the M2 phenotype. During the induction of RAW264.7 cells toward M1 polarization, luteolin intervention inhibited the release of pro-inflammatory factors and promoted the release of anti-inflammatory factors, suppressed nuclear translocation of NF-κB p65/p50, promoted IκBα expression while inhibiting its phosphorylation, and inhibited NF-κB p65 expression and its phosphorylation. These findings indicate that luteolin can regulate macrophage polarization through the NF-κB signaling pathway, showing potential therapeutic effects on knee osteoarthritis.

Key words: luteolin, macrophage, nuclear factor-κB signaling pathway, network pharmacology, knee osteoarthritis, macrophage polarization 

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