基于HPLC-MS的黄芪补肾活血汤调控ALOX15/GXP4通路预防芳香化酶抑制剂所致骨质疏松的机制研究OA

Objective To explore the active components of Huangqi Bushen Huoxue Decoction(黄芪补肾活血汤,HQD)and its mechanism of preventing osteoporosis caused by aromatase inhibitors(AIs).Methods High performance liquid chromatography-mass spectrometry(HPLC-MS)was used to identify the composition classification and active components of HQD freeze-dried powder.In vivo experiment,C57BL/6J female mice aged 6-8 weeks were randomly divided into sham operation group,model group,high,medium and low doses of HQD and positive control group,with 10 mice in each group.Three months later,the bone mineral density and bone microstructure of femur were detected by Micro-CT,the pathological morphology of femur was detected by hematoxylin-eosin staining,the levels of E2,BALP and OT/BGP in serum were detected by ELISA,and the expressions of ALOX15,GXP4 and ACSL4 in bone tissue were detected by immunohistochemistry.Results 45 kinds of drugs were identified by HPLC-MS,and there were 283 secondary active components.The main components were psoralen,luteolin,catechol,cholic acid aldehyde and quercetin.Compared with the model group,the experimental results in vivo showed that the femoral bone density increased,the bone microstructure improved,the tra-becular bone became thicker,denser and more numerous,and the contents of E2,BALP and OT/BGP in serum increased significantly.Immunohistochemical results showed that the expressions of ALOX15 and ACSL4 protein in bone tissue increased,while the expression of GPX4 protein decreased.Conclusion This study comprehensively clarified the active components in HQD,and revealed that this pre-scription may improve the estrogen level,promote osteoblast-mediated bone formation and improve bone remodeling by regulating the ALOX15/GPX4 iron death signal pathway of osteoblasts,thus preventing the occurrence and development of osteoporosis caused by AIs.