补肾壮骨汤调控Nrf2/HO-1信号通路抑制铁死亡改善2型糖尿病性骨质疏松模型的机制研究OA

Objective To identify the effective components and mechanism of Bushen Zhuanggu Tang for the treatment of type 2 diabetic osteoporosis(T2DOP)using a comprehensive strategy including liquid chromatography/mass spectrometry(LC/MS)and network pharmacology,combined with experimental verification.Methods The effective chemical components in Bushen Zhuanggu Tang were analyzed and identified by ultra-high-performance LC-Q-Orbitrap-MS.Genes related to T2DOP and ferroptosis were retrieved from the database,and the intersection was obtained to identify potential targets of the active ingredients for the treatment of T2DOP by regulating the ferroptosis pathway.A T2DOP rat model was induced by intraperitoneal injection of streptozotocin combined with high-sugar and high-fat feeding,and the rats were then administered Bushen Zhuanggu Tang for 8 weeks.Body weight,blood glucose,and glycosylated hemoglobin were measured.Bone morphological changes were detected by Micro CT.Total iron ion,superoxide dismutase(SOD),and malondialdehyde(MDA)levels were detected using appropriate kits.Nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling-and ferroptosis-related proteins in bone tissue were detected by Western blot.Results A total of 848 targets of 46 identified compounds were predicted in the blood compounds of Bushen Zhuanggu Tang.Comparing the GeneCards,Comparative Toxicogenomics,and FerrDb V2 databases finally indicated 81 potential targets of Bushen Zhuanggu Tang against T2DOP by regulating ferroptosis.Protein-protein interaction network analysis indicated that silent information regulator 1,mammalian target of rapamycin,signal transducer and activator of transcription 3,Nrf2,SRC,epidermal growth factor receptor,interleukin(IL)-6,mitogen-activated protein kinase-3,IL-1B,peroxisome proliferator-activated receptor γ,albumin,and other nodes may be important targets for Bushen Zhuanggu Tang in the treatment of T2DOP.Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis identified 134 signaling pathways related to the intersection target genes.The main pathways were metabolic pathways,lipid and atherosclerosis,and ferroptosis signaling pathways in diabetic complications.The in vivo experiments showed that the body weight of rats in the Model group increased slowly compared with the Control group,while blood glucose and glycosylated hemoglobin levels increased,trabecular bone was destroyed,bone mineral density,bone volume fraction,and trabecular thickness and density decreased,the degree of trabecular separation increased,total iron ion and MDA levels in bone tissue increased,SOD activity decreased,and Nrf2,HO-1,and ferroptosis-related protein levels increased(P<0.05).Compared with the Model group,weight gain increased,blood glucose and glycosylated hemoglobin levels decreased,destruction of the bone trabecula decreased,bone mineral density,bone volume fraction,and bone trabecula number and density increased,total iron ion and MDA levels in bone tissue decreased,while SOD activity increased,and Nrf2,HO-1,and ferroptosis-related protein levels decreased(P<0.05).Conclusions Bushen Zhuanggu Tang has multi-component,multi-target,and multi-pathway characteristics for the treatment of T2DOP.The result of in vivo experiments show that Bushen Zhuanggu Tang can prevent and treat T2DOP by regulating the Nrf2/HO-1 signaling pathway and inhibiting oxidative stress and ferroptosis.