Study On The Pharmacodynamic Material Basis And Network Pharmacology Of Modified Danggui Buxue Decoction In Treating Leukopenia

Objective: This study was conducted to investigate the material basis and potential mechanism of action of Modified Danggui Buxue Decoction(MDBD)in treating leukopenia based on the in vitro chemical composition study of MDBD,combined with serum pharmacochemistry and network pharmacology.Methods: Firstly,a database of chemical components related to MDBD was established through literature research,and the mass data of MDBD were collected by ultra-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry(UPLCQ-TOF-MS),combined with UNIFI platform and molecular network technology to identify known components and unknown components in MDBD,and all chemical components were attribution to the individual herbs.Secondly,serum samples were collected from the rat hepatic portal vein after gavage with MDBD.A study of serum pharmacochemistry was performed using UPLC-Q-TOF-MS technology in combination with UNIFI data analysis platform,the blood prototype components and metabolites of MDBD in rat serum were screened,and the possible metabolic pathways were analyzed in combination with the chemical composition identification results.Finally,on the basis of blood prototype components,the network pharmacology was used to predict the action targets of MDBD,and the targets related to leukopenia were screened for protein-protein interaction network(PPI)construction and enrichment analysis,and a blood component-target-pathway network was constructed.The binding activity of core targets and blood components was also validated using molecular docking techniques.Results: Based on the established method,a total of 208 chemical components,including flavonoids,saponins,organic acids,phthalides and other compounds,were identified in MDBD in positive and negative ion modes,of which 35 were accurately identified by reference standards.30 blood prototype components and 52 metabolites were identified in rat serum.Blood components in MDBD are mainly flavonoids and saponins,and the metabolic pathways in vivo are mainly reduction,oxidation,sulfation and methylation,etc.The network pharmacology predicted 514 targets of blood components of MDBD,180 targets related to leukopenia disease,and screened 8 core targets such as TP53,STAT3,TNF and IL-6 etc.MDBD plays a role in treating leukopenia mainly through signaling pathways such as PI3K-Akt signaling pathway,cellular senescence and Fox O signaling pathway.The molecular docking results suggested good binding activity between some of the blood components of MDBD and the core targets,indicating that the network pharmacological results are reliable.Conclusions: In this study,the in vivo and in vitro chemical constituents of MDBD were comprehensively and systematically identified using multiple data collection modes and data analysis platforms,and the main active ingredients of MDBD in treating leukopenia were predicted to be Genistein,Isoastragaloside II,6-Methoxycoumarin,and Astramembrannin II,etc.The mechanism may be related to the regulation of signaling pathways such as PI3K-Akt signaling pathway,cellular senescence and Fox O signaling pathway.The present study initially elucidated the pharmacological substance basis and potential mechanism of action of MDBD in the treating leukopenia,and also provided a data reference for the subsequent studies to the MDBD.