Study On The Mechanism Of Peach Seed-safflower Combined With Folic Acid Against Coronary Atherosclerosis Based On Network Pharmacology And Its Clinical Target Gene Prediction

ObjectiveAtherosclerosis is a common pathological change in vascular diseases.It occurs in many places and involves many systems,such as cardio-cerebrovascular,neck blood vessels,lower limb blood vessels and renal blood vessels.Furthermore,coronary atherosclerosis is the most important health burden in the whole population,with a high prevalence base and the highest fatality rate.With the development of minimally invasive surgery techniques for heart stents,the cure rate of acute myocardial infarction caused by coronary atherosclerosis is significantly higher than before,but there are still many complications to be solved.Therefore,from the perspective of secondary prevention to delay and prevent coronary atherosclerosis,there is still a further exploration of significance.In recent years,the combined use of traditional Chinese medicine and western medicine in the treatment of coronary atherosclerosis has proved to be more effective than the single drug.Traditional Chinese medicine of peach seed and safflower is essential for promoting blood circulation and removing blood stasis as well as promoting qi and relieving pain.It is often used in the prescription for prevention and treatment of coronary atherosclerosis.The western medicine folic acid is also an important auxiliary drug in the guidelines for prevention and treatment of coronary atherosclerosis.This project intends to analyze the risk prediction targets of coronary atherosclerosis through gene expression profiling from the perspective of bioinformatics,and explore the effect targets and pathways of the combined prevention and treatment of coronary atherosclerosis from the perspective of network pharmacology,so as to further understand the specific molecular mechanism.Methods1.Bioinformatics target gene predictionIn this study,we obtained GSE90074 gene chip data from the gene expression integration(GEO)database.First,differentially expressed genes(DEGs)were screened by ratio analysis between the lesion group and the normal group,and functional cluster analysis was performed according to the gene ontology(GO)and the Kyoto encyclopedia of genes and genomes(KEGG).Secondly,through STRING database,protein interaction map was established to understand the relationship between different genes.Thirdly,important modules and key genes are screened by Cytohubba and MCODE.Finally,the clinical data of gene chip was mined to establish a multivariate logistic regression model between clinical characteristics and key gene expression,so as to predict the risk relationship between target genes and coronary atherosclerosis.2.Network pharmacology analysisIn the network pharmacology analysis of the mechanism of action of peach kernel and safflower combined with folic acid against coronary atherosclerosis,the effective components of peach kernel and safflower were firstly obtained through TCMSP database,and the structure was obtained by searching folic acid in Pubchem database.Secondly,corresponding target genes were obtained from the UniProt database and PharmMapper database platforms,and the corresponding target genes of peach seed,safflower and folic acid were combined and deduplicated.Thirdly,disease-related targets were searched in OMIM and GeneCards databases with “coronary atherosclerosis” as the key word,and the genes corresponding to the disease were intersected with the drug target genes to obtain the target genes possibly corresponding to the anti-coronary atherosclerosis effects of peach seed and safflower combined with folic acid,so as to construct a drug regulatory network.Finally,the functional enrichment analysis was carried out on the target genes of peach kernel and safflower combined with folic acid in the anti-coronary atherosclerosis,and the important modules and key genes were selected.Results1.Bioinformatics target gene prediction resultsThrough the comprehensive analysis of the gene chip for obstructive coronary atherosclerosis,432 differentially expressed genes were obtained,of which 81 were up-regulated and 96 were down-regulated.Up-down-regulated gene function enrichment analysis showed that the main molecular functions were closely related to endogenous or exogenous lipid antigen binding,ligase ubiquitin-protein ligase,ligase zinc ion and androgen receptor binding,and involved in the positive regulation of endothelial cell apoptosis,the regulation of immunoglobulin secretion and the metabolic processes such as immune response.The protein interaction network of differential genes was mapped,and important modules and key genes were screened out to obtain 11 key genes :BTRC,NFKBIA,PSMB1,PSMC1,COPS5,NEDD8,PSMA6,PSMD4,PSMD12,UBC,RPS4 X.Based on the clinical characteristic data,a model was established to predict the risk of OCAD in relation to gender,history of high fat,NEDD8,BTRC,COPS5,PSMA6,PSMD12,RPS4 X and NFKBI expression levels.2.Results of network pharmacology analysisThe molecular structure of 45 active components of peach kernel and safflower and folic acid were obtained.Combined with coronary atherosclerosis disease,the targets of peach seed and safflower combined with folic acid against coronary atherosclerosis were analyzed,and a drug regulatory network was constructed.It shows the interaction network between the target genes and the target genes of folic acid,peach seed and safflower,as well as the relationship between folic acid,peach seed and safflower and the pharmacophore of the target genes,so as to understand the relationship between the drugs.The network interaction relationship between the targets of drug action against coronary atherosclerosis was constructed,and its functional enrichment,important modules and key targets were analyzed.Important modules GO enrichment analysis,mainly involved in DNA templating transcription and transcription positive regulation;Negative regulation of apoptosis process;Promoting positive regulation of RNA polymerase II transcription;Development of prostatic epithelial cell differentiation;Modulating the response to DNA damage stimuli;Its molecular function is mainly to act on transcription factor activity,and sequence-specific DNA binding.Receptor signaling protein tyrosine kinase activity;Steroid combination.KEGG enrichment analysis of important modules,Mainly included thyroid hormone signaling pathway,p53 signaling pathway,PI3 K signaling pathways-Akt signaling pathways,HIF-1,cell apoptosis,MAPK signaling pathways,Toll-like receptors signaling pathways,cell cycle and Wnt signaling pathways,nonalcoholic fatty liver disease(NAFLD),hepatitis b,tuberculosis,whooping cough,legionnaires’ disease,toxoplasmosis,herpes simplex infection,infection of HTLV-I,swine flu,viral myocarditis,salmonella infection,amoebiasis,epstein-barr virus infection,ErbB signaling pathways,tumor necrosis factor signaling pathways,prolactin signaling pathways,FoxO Signaling pathway,nodal receptor signaling pathway,sticky spot chagas disease,multiple cancer pathways.Finally,eight key targets were obtained,including ALB,MAPK8,EGFR,CASP3,VEGFA,MYC,ESR1 and CCND1.ConclusionsIn this project,network pharmacology and bioinformatics technology were applied to comprehensively analyze coronary atherosclerosis,so as to understand the effect targets and pathways of peach seed and safflower combined with folic acid in the anti-coronary atherosclerosis from the molecular target level.The mechanism of action of peach seed and safflower combined with folic acid against coronary atherosclerosis was preliminarily explained,which may interfere with the function of proteins or enzymes by affecting the process of DNA binding,modification,damage,transcription,translation into proteins and protein modification.Moreover,it was found that many of the pathways of peach seed-safflower combined with folic acid in the anti-atherosclerosis of coronary arteries were related to various cancer pathways.