Mechanism Of Danshen In Treating Ulcerative Colitis Based On Network Pharmacology Approach

Objective:Danshen(Radix et Rhizoma Salviae Miltiorrhizae)has been widely used in the clinical treatment of colitis,but its drug material basis and the action mechanism are currently unclear.This study will apply network pharmacology research technology to explore the potential effective components of Danshen in the treatment of ulcerative colitis-the target of action-signaling and collaterals,and further verify the protective effect of Danshen on ulcerative colitis and the accuracy of related pathways through in vivo experiments.The action mechanism of Danshen in the treatment of ulcerative colitis is systematically and comprehensively illustrated,which providing a factual basis for its clinical application.Methods:(1)Chinese Medicine System Pharmacology Database and Analysis Platform(TCMSP),Traditional Chinese Medicine Integrated Database(TCMID),Chemistry Specialty Database-Traditional Chinese Medicine and Active Ingredient Database,ETCM Database,etc.and literature were used to obtain the chemical components of Salvia miltiorrhiza.Kinetic ADME screens the active ingredients of Salvia miltiorrhiza,and then the SwissTarget Prediction platform was applied to predict the target of the active ingredients of Salvia miltiorrhiza.(2)GeneCards,DRUGBANK,OMIM,DisGeNET,TTD,MalaCards and other databases were applied to obtain ulcerative colitis disease targets.Venny 2.1.0 online drawing software was applied to obtain the intersection target between the potential targets of Salvia miltiorrhiza and the target of ulcerative colitis disease,and then STRING protein database was applied to construct a protein-protein interaction(PPI)network and explore the core functional module.(3)Perform Gene Ontology(GO)and KEGG enrichment analysis on the intersection targets through Metascape,and use Cytoscape 3.7.1 software to visualize the "drug-active ingredient-target point-pathway-disease" network.It preliminary revealed the potential mechanism of Danshen in the treatment of ulcerative colitis.(4)The protective effect of Danshen on ulcerative colitis was verified through the animal model of colitis.The specific method is as follows:Select 30 male C57BL/6J mice(20±2g)and randomly divide them into three groups(10 mice/group):blank control group,model group(4%DSS),and administration group(4%DSS)+455mg/kg/d Danshen granules).The mice were given by intragastric administration for 7 days while building the model,weighing and recording the weight changes of different groups of mice every day,observing the mental state of the mice,stool characteristics(loose stool,diarrhea,fresh blood stool),performing fecal occult blood detection and calculating the disease activity index(disease activity index,DAI)score.On the 8th day of model building,blood was collected and the mice were sacrificed,the colon was separated,the general shape of the colon was observed and its length was measured.Through pathological examination,the effect of the drug on the structure of the mouse colon was evaluated.Enzyme-linked immunosorbent assay(ELISA)detects serum tumor necrosis factor-α(tumor necrosis factor,TNF-α),interleukin-1β(interleukin-1 beta,IL-1β),and interleukin-6(interleukin-6,IL).-6)The level of interleukin-10(IL-10)to detect the effect of drugs on inflammation in mice with colitis.Results:(1)A total of 42 effective active ingredients of Salvia miltiorrhiza were predicted by network pharmacology analysis,and the core active ingredients for the treatment of ulcerative colitis were luteolin,y-sitosterol,caffeic acid,etc.;(2)Potential target of Salvia miltiorrhiza 152 points,788 targets for ulcerative colitis disease,and 43 intersection targets,mainly acted on core targets PTGS2,STAT3,AKT1,SRC,EGFR,ESR1,MMP9,PPARG,MMP2,AHR,KDR,CYP19A1 MMP1,MPO,MET,ABCG2,IGF1R,MMP3,etc.;(3)Salvia miltiorrhiza regulating signal pathway enriches a total of 122 KEGG pathways,including arachidonic acid metabolism,interleukin-17 signaling pathway,TNF signaling pathway and other inflammatory signaling pathways.It may play an important role in the anti-ulcerative colitis process of Danshen.(4)The results of the colitis animal model verifying the efficacy of Danshen showed that compared with the blank group,the mice showed significant weight loss,diarrhea,mucus and blood in the stool 4 days after modeling,and the Danshen granule aqueous solution(455mg/kg)could be significantly reduced.It also relieved the symptoms of DSS-induced ulcerative colitis model mice,improved the shortening of the mouse colon,reduced the infiltration of inflammatory cells in the colon and actively hyperplasia of the glands,The detection of inflammatory indexes showed that Danshen could significantly inhibit the increase of TNF-α and IL-6 caused by DSS,and up-regulate the level of IL-10 in serum,the difference was statistically significant(P<0.05).The above results indicate that Danshen has a significant effect on ulcerative colitis.Conclusion:Danshen may regulate ulcerative colitis through multiple components,targets,and pathways.It is speculated that it mainly mediates arachidonic acid metabolism,relaxin signaling pathway,interleukin-17 signaling pathway,focal adhesion,platelet activation,Th17 cell differentiation and other signaling pathways to regulate inflammation,immunity,apoptosis,and tissue remodeling.Focal adhesion,coagulation cascade,reactive oxygen metabolism and other biological processes perform the actions of anti-inflammatory,immune regulation,repair of the intestinal mucosal barrier,anticoagulation,anti-fibrosis,anti-oxidation,etc.It was further verified by an animal models with colitis that Danshen can treat ulcerative colitis by inhibiting TNF-α,IL-6 and other pro-inflammatory mediators and increasing the level of anti-inflammatory mediator IL-10.