关键词: 四黄止痢颗粒; 畜禽腹泻; UPLC-Q-TOF-MS; 网络药理学; 分子动力学模拟

Abstract: 【Objective】 This study was conducted to systematically investigate the pharmacological mechanisms of Sihuang Zhili granules against diarrheal diseases in livestock and poultry,and provide a theoretical basis for optimizing clinical diarrhea management strategies. 【Method】 The main active ingredients of Sihuang Zhili granules were analyzed using UPLC-Q-TOF-MS technology.Active ingredients were screened and their potential targets were predicted via SwissTargetPrediction database.Disease targets related to livestock and poultry diarrhea were identified through GeneCards and OMIM databases.An active component-target network was constructed using Cytoscape 3.10.3 software.The intersection between Sihuang ZhiLi granules’ targets and diarrhea-related targets was identified and visualized using a Venn diagram.A protein-protein interaction (PPI) network was subsequently built using Cytoscape 3.10.3. GO functional annotation and KEGG pathway analysis of intersection genes was performed using DAVID database.Finally,molecular docking was executed with AutoDock Vina to validate the binding activity between key components and core targets,followed by 100 ns molecular dynamics simulations conducted with Gromacs 2022 software. 【Result】 Through UPLC-Q-TOF-MS, 88 active ingredients were identified in Sihuang Zhili granules. Subsequently, 16 key components were screened out based on literature validation and analysis using SwissTargetPrediction database. The database analysis revealed that there were 289 potential targets for the drug,4 567 targets related to livestock and poultry diarrhea,and 201 intersecting targets.In terms of biological process,GO functional annotation was mainly related to protein phosphorylation,negative regulation of apoptosis,and response to exogenous stimuli.In terms of cell component,it was mainly related to neuronal cell body,plasma membrane,cytoplasm,etc.In terms of molecular function,it was mainly related to protein binding,ATP binding,and protein-containing complex binding.KEGG enrichment analysis mainly involved phosphatidylinositol signaling system,protein processing in endoplasmic reticulum,tyrosine metabolism,PI3K-Akt signaling pathway,FoxO signaling pathway,Ras signaling pathway,VEGF signaling pathway,etc.The molecular docking results showed that the binding energy of the top 10 core targets and the 6 main active ingredients of Sihuang Zhili granules was less than －5 kJ/mol,indicating that the docking effect was good.Among them,acacetin had the lowest binding energy to heat shock protein (7D1V),which was －9.4 kJ/mol.100 ns molecular dynamics simulation further confirmed that acacetin had good binding stability and activity with 7D1V. 【Conclusion】 The prediction results showed that Sihuang Zhili granules acted on tumor necrosis factor (TNF),heat shock protein 90α family class A member 1 (HSP90AA1),rapamycin target protein (mTOR),estrogen receptor 1 (ESR1) and other core targets through key active ingredients such as norwogonin,wogonin,kaempferol,5,7,2',6'-tetrahydroxyflavone,acacetin,and oroxylin A,and other core targets,through tyrosine metabolism,PI3K-Akt signaling pathway,FoxO signaling pathway,phosphatidylinositol signaling system and other pathways played a role in the treatment of livestock and poultry diarrhea.

Key words: Sihuang Zhili granules; livestock and poultry diarrhea; UPLC-Q-TOF-MS; network pharmacology; molecular dynamics simulation