To explore the therapeutic mechanism of Xianfang Huoming decoction in sepsis based on UPLC-Q-TOF-MS/MS combined with bioinformatics and network pharmacology

Wang Jie; Li Miaomiao; Qi Yanan; Zhou Tian; Wang Ting; Xu Yaqian; Liu Cheng

doi:10.3969/j.issn.1002-1949.2025.09.003

Abstract：Objective To explore the therapeutic mechanism of Xianfang Huoming decoction in sepsis based on ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS)combined with bioinformatics and network pharmacology.Methods The active ingredients of Xianfang Huoming decoction were analyzed by using UPLC-Q-TOF-MS/MS.The active ingredients obtained by the analysis and those retrieved through literature search were imported into traditional Chinese medicine systems pharmacology(TCMSP)databases to screen the effective active ingredients and potential targets of Xianfang Huoming decoction.The differentially expressed genes of sepsis were analyzed by GEO database.The GeneCards and OMIM databases were used to screen the targets of sepsis.The Venny tool was used to screen the drug-disease mapping targets,and the String database was used to construct the protein-protein interaction(PPI)network of overlapping targets,which was then imported into the Cytoscape software to establish the drug-ingredient-target-disease network diagram,and to screen out the core targets and key components.Metascape database was used to analyze the KEGG pathway and GO functional enrichment,and the signaling pathways with higher relevance were screened based on the P value.The molecular docking and molecular dynamics simulations of the key active ingredients and core targets were carried out by AutoDock,Pymol,Gromacs and other software to further elucidate the stability of the binding of the key ingredients to the targets at the molecular level.Results 39 compounds were analyzed by UPLC-Q-TOF-MS/MS,and the obtained compounds were imported into the TCMSP database and further screened under the conditions of"OB≥30％,DL≥0.18"to obtain 24 active ingredients.We obtained 177 potential targets of Xianfang Huoming decoction for the treatment of sepsis.PPI network analysis screened the top 5 core targets based on the degree value,which were interleukin(IL)-6,tumor necrosis factor(TNF),signal transducer and activator of transcription 3(STAT3),tumor suppressor factor P53(TP53),and protein kinase B(AKT)type 1.GO and KEGG enrichment analyses showed that Xianfang Huoming decoction involved multiple signaling pathways in sepsis therapy,mainly including mitogen-activated protein kinase(MAPK),PI3K-Akt.Through molecular docking and molecular dynamics simulation,it was found that quercetin,luteolin,β-sitosterol and naringenin,the top 4 core components of Xianfang Huoming decoction,had good binding ability with TP53,AKT1,STAT3,IL-6 and TNF.Conclusions Xianfang Huoming decoction has a"multi-component,multi-target,multi-pathway"effect.Quercetin,luteolin,β-sitosterol and naringenin may be the main active components of Xianfang Huoming decoction,and TP53,AKT1,STAT3,IL-6 and TNF may be the potential therapeutic targets of Xianfang Huoming decoction in the treatment of sepsis.Xianfang Huoming decoction may exert therapeutic effects through signaling pathways such as MAPK,PI3K-Akt.