Screening Of Active Components Of Bushen Huoxue Decoction Against Breast Cancer Bone Metastasis Based On Network Pharmacolog

Objective: To investigate the key components of the blood-entering component of tonifying and invigorating blood soup against bone metastasis by screening for anti-tumour active ingredients.Methods: Bioinformatics analysis was carried out through network pharmacology to construct a "Chinese herbal medicine-blood component-target-pathway" diagram to identify the components of the blood component that interact with breast cancer bone metastasis.The effect of the relevant components on the apoptosis and cycle of breast cancer cells was detected.Results: A total of 731 targets were obtained from Swiss Target Prediction database,and 382 targets were obtained by merging and deleting duplicates.The Gene Cards database was used to obtain 7800 gene targets related to bone metastasis of breast cancer.Using the Lianchuan Bio Cloud platform to map the targets of Kidney and Blood Tonic and breast cancer bone metastasis to each other,276 potential targets with therapeutic effects were obtained.The potential targets were imported into Cytoscape 3.9.0 software to obtain the protein-protein interaction network,and the enrichment analysis of GO and KEGG pathways of potential targets was performed,which revealed that they were involved in cell proliferation,apoptosis,protein synthesis,metabolism and regulation of key cellular responses in cell cycle and osteoblast regulation,including OPG/RANK/RANKL pathway,PI3K/AKT signaling pathway,MAP/AKT signaling pathway,OPG/AKT signaling pathway,and OPG/AKT signaling pathway./AKT signaling pathway,MAPK signaling pathway,estrogen signaling pathway,tumor necrosis factor signaling pathway,m TOR signaling pathway,NF-κB signaling pathway,etc.,which are involved in the regulation of different aspects of the pathological process of breast cancer bone metastasis.The above screened pathways and pathway-related targets were used to construct a "Chinese herbal medicine-blood-entry component-target-pathway" map using Cytoscape 3.9.0software,and the remaining 13 chemical components were further screened by excluding chlorogenic acid,which had no obvious interaction with them.Through two CCK8 experiments,the components with better anti-tumor effects were imperatorin,osthole,and isopsoralen;three components were found to have no significant effects on the cell cycle of four types of breast cancer by flow cytometry.Imperatorin,osthole,and isopsoralen were unable to induce apoptosis in MDA-MB-231 cells,and the induction of apoptosis in ZR-75-1 was not significant,while in SK-BR-3(imperatorin 73.2%,osthole 71.5%,and isopsoralen 85.3%),MCF-7 cells(imperatorin49.4%,osthole 47.2%,and isopsoralen 57.2%),the pro-apoptotic effect of isopsoralen was better,and the pro-apoptotic effect of imperatorin was similar to that of osthole.Conclusion: The 14 monomeric compounds of guanosine,monosidine,loganin,psoralenoside,isopsoralenoside,imperatorin,psoralen,isopsoralen,angelol K,angelol D,angenomalin,neobavaisoflavon,isoneobavaisoflavone,and osthole interacted with bone metastasis of breast cancer through biological analysis of tonic kidney and blood soup by network pharmacology;the13 monomeric compounds that were available were The CCK8 assay was performed on breast cancer MDA-MB-231,ZR-75-1,MCF-7 and SK-BR-3,and imperatorin,osthole and isopsoralen were the components with better anti-tumor effects;the cycle and apoptosis assay was performed by flow cytometry,and isopsoralen induced the best apoptosis,and imperatorin and osthole had similar degrees of pro-apoptotic effects;combined with the experimental Combined with the experimental results,it was considered that imperatorin and isopsoralen were the key bloodentering components of the tonic kidney and blood invigorating soup,which originated from two Chinese herbal medicines,duhuo and psoralea corylifolia,in line with the TCM theory of tonifying the kidney and strengthening the bones,and had obvious anti-tumour and antiosteoporosis effects.