目的 基于血清非靶向代谢组学、血清药物化学和网络药理学，探究补肾通络方治疗膝骨关节炎（KOA）的药效物质基础和代谢调控机制。方法 采用单碘乙酸钠（MIA）诱导大鼠KOA模型，将实验大鼠随机分为对照组、模型组和补肾通络方低、高剂量（400、800 mg·kg^-1）组。采用ELISA检测大鼠血清炎症因子水平；通过苏木素-伊红（HE）染色和番红O-固绿染色观察膝关节软骨组织形态，并依据国际骨关节炎研究学会（OARSI）半定量评分系统评估软骨退变程度。利用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱技术，鉴定大鼠血清药物移行成分、筛选潜在差异代谢物并开展代谢通路富集分析；通过偏最小二乘回归法建立方剂谱效关联模型，明确核心药效物质。结果 补肾通络方可显著降低MIA诱导的KOA大鼠软骨损伤，显著抑制机体炎症反应。代谢组学共筛选出120个差异代谢物，主要涉及氨基酸代谢、脂质代谢相关的6条通路；从大鼠血清中鉴定出18个补肾通络方入血移行成分。谱效关联分析发现，甜菜碱、宝藿苷I和地黄苦苷元为该方发挥抗炎作用的核心药效物质，并构建补肾通络方治疗KOA的有效成分-靶点-代谢物-通路的关联网络。结论 补肾通络方能够显著改善KOA大鼠的软骨损伤和炎症反应。其作用机制可能与宝藿苷I、地黄苦苷元和甜菜碱通过调控PTGS2和ALOX5基因的表达，调节血清中亚油酸、鞘氨醇-1-磷酸、棕榈烯酸、磷脂酰酸、棕榈酸等不饱和脂肪酸含量，进而调控亚油酸和花生四烯酸等代谢通路有关。

Objective Based on serum non-targeted metabolomics, serum pharmacokinetics, and network pharmacology, to explore the pharmacodynamic substance basis and metabolic regulatory mechanism of Bushen Tongluo Formula in the treatment of knee osteoarthritis (KOA). Methods A rat model of KOA was induced by sodium moniodoethanoate (MIA), and the experimental rats were randomly divided into the control group, the model group, and the low and high-dose (400 and 800 mg·kg^-1) groups of the Bushen Tongluo Formula. The levels of inflammatory factors in the rat serum were detected by ELISA; the morphology of knee joint cartilage tissue was observed by hematoxylin-eosin (HE) staining and safranin O-fast green staining, and the degree of cartilage degeneration was evaluated according to the semi-quantitative scoring system of the International Osteoarthritis Research Society (OARSI). The serum drug transport components of rats were identified by ultra-high performance liquid chromatography-tandem quadrupole/electrostatic field orbitrap high-resolution mass spectrometry, potential differential metabolites were screened, and metabolic pathway enrichment analysis was conducted; the formula-spectrum-effect association model was established by partial least squares regression to clarify the core pharmacodynamic substances. Results Bushen Tongluo Formula could significantly reduce cartilage damage in MIA-induced KOA rats and significantly inhibit the inflammatory response. A total of 120 differential metabolites were screened out, mainly involving six pathways related to amino acid metabolism and lipid metabolism; 18 serum transport components of the Bushen Tongluo Formula were identified. The spectrum-effect association analysis revealed that betaine, baohuoside I, and rehmapicrogenin were the core pharmacodynamic substances of this formula for anti-inflammatory effects, and the Bushen Tongluo Formula for the treatment of KOA was constructed as an association network of components, targets, metabolites, and pathways. Conclusion Bushen Tongluo Formula can significantly improve the cartilage damage and inflammatory response in KOA rats. Its mechanism of action may be related to the regulation of the expression of PTGS2 and ALOX5 genes by betaine, baohuo glycoside I, and rehmapicrogenin, which in turn regulates the content of unsaturated fatty acids such as linoleic acid, sphingosine-1- phosphate, palmitoleic acid, phosphatidic acid, and palmitic acid in the serum, thereby regulating metabolic pathways such as linoleic acid and arachidonic acid.

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国家自然科学基金资助项目（82104667）；山西省科技厅基础研究项目（202203021211228）；山西省中医药重点研究室建设项目（zyyyjs2024031）；山西省中医药创新团队（zyytd2024008）；山西省卫健委强基工程科技兴医创新计划项目（2026QJGC024）