Integrated Metabolomics And Network Pharmacology To Reveal The Pharmacodynamic Substance Of Huashi Baidu Formula

Huashibaidu formula(HSBDF)is composed of 14 traditional Chinese medicines(TCMs),including Ephedrae Herba(MH),Semen Armeniacae Amarum(KXR),Glycyrrhizae Radix et Rhizoma(GC),Pogostemonis Herba(GHX),Magnoliae Officinalis Cortex(HP),Atractylodis Rhizama(CZ),Tsaoko Fructus(CG),Pinelliae Rhizoma(BX),Poria(FL),Radix Padoniae Rubra(CS),Rhei Radix et Rhizoma(DH),Astragali Radix(HQ),Descurainiae Semen(TLZ)and Gypsum Fibrosum(SG).This formula exerts therapeutic effects through functions such as removing dampness,dispersing the lungs,promoting blood circulation,and detoxifying the body.Currently,research on HSBDF primarily focuses on predicting possible active ingredients and investigating the pharmacological mechanisms using network pharmacology and molecular docking technology.Furthermore,there have been analyses and research on its therapeutic effects through review and meta-analysis.However,there are still deficiencies in the basic research of its chemical substances.The research on blood components has not been reported,and the understanding of its pharmacological substances and their action mechanisms remains unclear.In order to elucidate the material basis,pharmacological substances,and possible mechanisms of action of HSBDF,this study aims to systematically characterize and analyze the composition and structure of HSBDF,identify the sources and types of compounds,and investigate its effects on blood components.By studying the exposure patterns and movement trajectories of HSBDF components in the bodies of rats after oral administration,this research aims to reveal the exposure of the prototype and related metabolites of HSBDF,providing a scientific basis for determining the effective component group.Finally,by using network pharmacology methods,the study focuses on the prototype blood components to preliminarily investigate the mechanism of HSBDF in the treatment of novel coronavirus pneumonia(COVID-19).This research aims to confirm the chemical and pharmacological substance basis of HSBDF,elaborate on the effective ingredients and mechanisms of action,provide scientific guidance and theoretical basis for its treatment of COVID-19,and serve as a reference for new drug development.In this study,UPLC-Q-TOF-MSE technology combined with UNIFI software and self-buit library were utilized to comprehensively characterize and analyze the chemical components in HSBDF.Firstly,a database comprising 2253 chemical components of HSBDF was established through the TCMIP-V2.0 database and systematic literature search,including their names,molecular formulas,and structural formulas.Next,the LC-MS conditions for HSBDF were optimized.The mass spectrometry fragmentation patterns and chromatographic retention behaviors of flavonoids,phenylpropanoids,terpenes,anthraquinones,alkaloids,and phenolic acids were investigated based on 104 standard samples and literature references.Subsequently,the imported HSBDF MS data were automatically compared and identified using the UNIFI software and a self-built database.The identification results from UNIFI were further validated by accurate mass number,characteristic ions and abundance,neutral loss,retention time,chromatographic peak strength,and information from the standard samples.Correct results were confirmed,and the structures that were incorrectly identified or lacked candidate structures were analyzed.A total of 620 chemical components were identified,including 104 compounds clearly identified by the standard samples.Among them,133 flavonoids,65 triterpenoid saponins,77 terpenes,47 glycosides,34 alkaloids,41 lignans,41 tannins,30 phenolic acids,23 anthraquinones,55 phenylpropanoids,13 phenylethanosides,20 fatty acids,9 oligosaccharides,5 pyranones,3 aminoacids,3 organic acids,3 nucleosides,5coumarins,2 stilbenes,and 11 others.144 from HP,56 from HQ,52 from CZ,105 from GHX,20 from CG,162 from GC,29 from BX,54 from TLZ,81 from MH,123 from CS,34 from KXR,114 from DH and 37 from FL.Collect SD rats blood samples at different time points after oral administration of the water and alcohol extracts of HSBDF,and systematically study their blood components and possible metabolites using UPLC-Q-TOF-MSE technology.On the basis of the method and results of the HSBDF component analysis,the blood components were analyzed using plasma blanks in conjunction with UNIFI software.Using both positive and negative ion modes,a total of 126 exposed components in the blood were identified,including 35 compounds that were confirmed by the standard samples.Among these,59 were prototype compounds introduced into the blood,82 were identified as metabolites,and 15 were found to be both prototypes and metabolites.The main metabolic processes involved hypoglycemia,glucuronidation,sulfonation,and others.The exposed components detected in the blood primarily originated from GC,MH,HP,CS,DH,and other sources.Finally,the online database was utilized to identify target proteins corresponding to the 59 prototype blood components and COVID-19.A mapping relationship between them was established,resulting in the identification of 292 potential targets.A protein-protein interaction(PPI)network was constructed,and 241 HUB genes were obtained.Based on this,GO function analysis and KEGG enrichment analysis were conducted.The results of the GO function analysis revealed that the molecular functions of the identified targets were primarily concentrated on transmembrane receptor protein tyrosine kinase activity,protein tyrosine kinase activity,scaffold protein binding,and others.The biological processes involved response to drugs,protein phosphorylation,response to lipopolysaccharides,and more.The cellular components were associated with membrane rafts,receptor complexes,focal adhesions,and other cellular structures.The findings from the KEGG enrichment analysis indicated that the therapeutic mechanism of HSBDF in treating COVID-19 was linked to cell survival and death,energy metabolism,and inflammatory reactions.The association network of the "component-target-function module" demonstrated that HSBDF could exert its therapeutic effect by regulating the expression of genes such as MAPK1,MAPK3,STAT3,SRC,AKT1,PTPN11,HSP90AA1,GRB2,RELA,and JUN.In summary,this study employed UPLC-Q-TOF-MSE technology in combination with UNIFI software and a self-built library to comprehensively analyze the chemical components of HSBDF,resulting in the identification of a total of 620 compound components.Subsequently,a detailed analysis was conducted on the blood components of HSBDF in rats.A total of 59 prototype blood components and 82 metabolites were characterized,with 15 compounds identified as both prototypes and metabolites.Lastly,network pharmacology was utilized to investigate the therapeutic mechanism of HSBDF in treating COVID-19,indicating potential involvement in cell survival and death,energy metabolism,inflammatory responses,and more.