基于血清药物化学和网络药理学探讨通腑止血方治疗脑出血的药效物质和作用机制OA

Objective To qualitatively analyze the blood components of Tongfu Zhixue formula by ultra-high performance liquid chro-matography tandem quadrupole electrostatic field orbit trap mass spectrometry(UPLC-Q-Exactive Orbitrap MS),and explore the po-tential mechanism of its treatment of intracerebral hemorrhage(ICH)by network pharmacology.Methods By comparing the chro-matograms of medicated serum and blank serum,combined with database secondary spectra,literature,and reference substance lysis information,the absorbed components of Tongfu Zhixue formula were analyzed and identified.The target genes of absorbed compo-nents and diseases were collected and screened using databases such as Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Swiss Target Prediction,Comparative Toxicogenomics Database,and Genecards,and network visua-lization processing was performed using Cytoscape software.Key targets were screened using a threshold of centrality≥1 252.14 and degree value≥37.94,and then a medicinal material-blood-entering component-key target diagram was constructed.The top 10 targets by degree value were taken as the core targets,and analyzed by Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrich-ment analysis and Gene Ontology(GO)enrichment analysis.The molecular docking of the top 10 core targets screened by degree value using Pymol software and their associated blood-entering components were verified.Results A total of 14 prototype components(ab-sorbed without metabolism)were identified,including ginsenoside Rf,ginsenoside Rb1,ginsenoside Rg1,ginsenoside F1,ginseno-side Rh1,dencichine,quercetin,citric acid,salicylic acid,aloe emodin,rhein,emodin,chrysophanol,and physcion.A total of 111 key targets were identified,and the top 10 core targets by degree value were PTGS2,IL-1β,TNF,CASP3,IL-6,HSP90AA1,Bcl-2,CYP3A4,RELA,and AKT1.The core targets were mainly enriched in TNF signaling pathway,IL-17 signaling pathway,and NF-κB signaling pathway.Molecular docking indicated that,apart from salicylic acid and citric acid,the binding energies of other core components(such as ginsenoside Rb1,rhein)with core targets were all≤-6.0 kcal/mol,and the binding energy of rhein with HSP90AA1 was-10.4 kcal/mol.Conclusion Tongfu Zhixue formula exerts its synergistic anti-inflammatory,anti-apoptotic,and vascular repair effects by inhibiting TNF/NF-κB inflammatory cascade and apoptotic pathways,activating PI3K-Akt pro-survival sig-naling pathway through seven absorbed components(ginsenoside Rb1,ginsenoside Rg1,aloe-emodin,rhein,emodin,chrysophanol,and physcion),which regulates core targets(e.g.,TNF,IL-1β,RELA,AKT1).