重骨颗粒通过调控Nrf2通路改善膝骨关节炎肝肾亏虚证患者的软骨损伤OA

Objective To explore the mechanism mediating the ameliorative effect of Chonggu Granules on cartilage injury in knee osteoarthritis(KOA).Methods Bioinformatics approaches were used for analyzing the active ingredients of Chonggu Granules,their potential targets,the KOA-and ferroptosis-related genes,and their intersection genes.A compound-ingredient-disease-target network and the protein-protein interaction(PPI)network were constructed to identify the key active ingredients and core targets,which were verified by molecular docking studies.In the clinical trial,62 KOA patients were randomly assigned into the observation group and control group(n=31)for treatment with glucosamine hydrochloride tablets and additional Chonggu Granules for 12 weeks,respectively,with 20 healthy participants serving as the healthy control group.Serum levels of cartilage injury markers(MMP-13,CTX-II,and COMP)and ferroptosis-related indices(GPX4,ACSL4,4-HNE,and Fe2+)were measured using ELISA,and expressions of Keap1,Nrf2,SLC7A11,HO-1,and GPX4 mRNAs were determined with real-time PCR.Results Four key active ingredients(quercetin,eupatilin,kaempferol,and wogonin)of Chonggu Granules and 5 core targets(AKT1,TP53,IL-6,JUN,and Nrf2)were identified,which were involved in cancer,AGE-RAGE signaling,and the NFE2L2(Nrf2)signaling pathways.Molecular docking showed strong binding between the active compounds and their targets,with NFE2L2 exhibiting the highest binding affinity.Compared with the control patients,KOA patients treated with Chonggu Granules showed significantly upregulated Nrf2,SLC7A11,HO-1,and GPX4 mRNA expressions(P<0.05),downregulated Keap1 expression(P<0.05),reduced serum levels of MMP-13,CTX-II,COMP,ACSL4,4-HNE,and Fe2+(P<0.05),and increased GPX4 level(P<0.05).Conclusion Chonggu Granules alleviate KOA through its key active ingredients quercetin and eupatilin,which activate the Nrf2 signaling pathway to modulate ferroptosis by targeting NFE2L2 and other therapeutic targets,suggesting a multi-component,multi-target,and multi-pathway synergistic mechanism for ameliorating chondrocyte injury in KOA.