Study On Mechanism Of Pulsatilla Decoction In Treating Chronic Ulcerative Colitis And Metabolism Of α-Hederin Saponins In Vitro And In Vitro

Objective: To find the potential biomarkers and metabolic pathways related to Pulsatilla Decoction in the treatment of chronic ulcerative colitis,to comprehensively elucidate the anti-colitis effect of Pulsatilla Decoction in combination with network pharmacological methods,and to discuss the effective ingredient of Pulsatilla Decoction α-chang The metabolites of ivy saponins in and outside the rat were identified and compared.Discuss related mechanisms of action from multiple dimensions.Methods: First,use the network pharmacology platform TCMSP,PubChem,Swiss Target Prediction Database,STICH Database,UniProt and TTD to screen the active compound targets of Pulsatilla Decoction,conduct network visualization analysis,and establish and analyze the relevant network with ulcerative colitis disease.Then,a mouse model of chronic ulcerative colitis was established,and Pulsatilla decoction was administered by gavage,and hematoxylin-eosin(HE)staining was used to verify the model.UHPLC-Q-TOF-MS/MS was used for data collection of biological samples,and Progenesis QI and SIMCA-P 14.1 software were used for data processing and analysis.In order to meet the conditions of P<0.05,FC≥1.2 or FC≤0.8,CV<30%(QC)and VIP>1 to find different compounds,identify the compounds according to their mass spectrometry information and online databases(HMDB,METLIN),and use the test The ROC curve verifies the accuracy of potential biomarkers,uses online software Metabo Analyst 4.0 for biomarker enrichment and metabolic pathway analysis,and uses Cytoscape software to construct a metabolite-gene-protein biological information network and protein Interaction network diagram.Finally,UHPLC-Q-TOF-MS/MS technology was used to analyze the metabolism of α-hedera saponins in SD rat plasma,urine,feces,bile and intestinal flora incubation samples according to the metabolite analysis strategy.The structure of the product was inferred,and the metabolic pathway of α-hedera saponins in the rat body was summarized.Results: Using the network pharmacology database,13 active ingredients of Pulsatilla were screened out,and 55 ingredient-disease targets were in total.The GO analysis and screening had significant differences P<0.05,and the first 15 biological process information,cell composition and molecular function information were taken.Network analysis shows that Pulsatilla,through a variety of active ingredients,acts on multiple key targets such as VEGFA,STAT3,CASP3,JUN,MAPK14,IL2,etc.,involving multiple signaling pathways such as vascular endothelial growth factor,PI3K-Akt,and tumor necrosis factor.Using metabonomics technology,40 different metabolites were identified between the chronic ulcerative colitis model group and the healthy group of rats,and 53 different metabolites were identified between the Pulsatilla decoction administration group and the model group of rats It predicts 10 potential biomarkers.The metabolic pathways involved mainly include galactose metabolism,arachidonic acid metabolism,glycerolipid metabolism,citrate cycle(TCA cycle),sphingolipid metabolism,glycolysis/sugar Xenobiotics,glyoxylic acid and dicarboxylic acid ester metabolism,pyrimidine metabolism,tyrosine metabolism,etc.A total of 28 metabolites were detected in vivo and in vitro after oral administration ofα-Hederinsaponins in rats,including 22 phase I and 6 phase II metabolites,involving multiple metabolic pathways.Conclusion: Combined with network pharmacology research,it revealed the mechanism of Pulsatilla Decoction in the treatment of ulcerative colitis,and provided theoretical support and scientific basis for further research on the pharmacological mechanism of Pulsatilla Decoction and its related preparations.Using metabolomics methods based on UHPLC-Q-TOF-MS/MS technology,searching for potential biomarkers and metabolic pathways related to the therapeutic effect of Pulsatilla Decoction,and constructing a comprehensive biological network from metabolites to genes and proteins These research results help to fully understand the biological mechanism of Pulsatilla Decoction in the treatment of chronic ulcerative colitis.According to the metabolite analysis strategy,this study identified the metabolites ofα-hedera saponins in and out of rats,and inferred the relevant metabolic pathways of α-hedera saponins in vivo,which is helpful to understand the clinical application of α-hedera Metabolic process in application.