CeRNA Network Of Danhong Injection In The Treatment Of Chronic Stable Angina Pectoris With Xue-yu Zheng

1 Research BackgroundRapid economic development,changes in lifestyle and dietary habits have continued to increase the level of risk factors for cardiovascular disease.As a common cardiovascular disease,stable angina pectoris usually occurs during activity or mental stress.It is caused by insufficient myocardial oxygen supply or myocardial It is caused by ischemia and is closely related to myocardial infarction,with a high incidence and a large social and economic burden.Stable angina pectoris is diagnosed as "chest arthralgia",and blood stasis syndrome is the main syndrome type.Physicians of past dynasties have accumulated a lot of practical experience in the treatment of chest arthralgia with drugs for promoting blood circulation and removing blood stasis.The treatment of angina pectoris with blood stasis syndrome has high safety and efficacy,so it is of great significance to explore the mechanism of Danhong injection in the treatment of chronic stability.With the development of high-throughput sequencing technology and bioinformatics technology,the research on TCM syndromes and compound pharmacology has entered the multi-omics level.Using the characteristics of omics big data,deep level,multiple indicators,and strong network correlation,it is possible to To explore the mechanism of Danhong injection in the treatment of chronic stable angina pectoris with blood stasis syndrome.2 Objective(1)Screening modules related to the efficacy of Danhong injection in the treatment of chronic stable angina pectoris with blood stasis syndrome;(2)To establish the ceRNA regulatory network related to Danhong injection in the treatment of chronic stable angina pectoris with blood stasis syndrome;(3)To explore the changes of Danhong injection at different time points in the treatment of chronic stable angina pectoris with blood stasis syndrome in the related modules of disease-syndrome efficacy and the molecular mechanism of ceRNA regulatory network.3 Contents and methodsThe 14-day and 30-day co-expression network of lncRNA,miRNA,and mRNA in patients with chronic stable angina pectoris and blood stasis syndrome treated with Danhong injection was constructed by weighted gene co-expression network analysis method,and the differential modules were identified,and the difference modules were calculated with the amount of Seattle angina pectoris in patients.The Pearson correlation coefficient of the score of angina pectoris attack frequency and the score of blood stasis syndrome,with the correlation coefficient greater than 0.25 and P<0.05 as the screening conditions,to identify the disease-syndrome efficacy-related modules at two time points.The Metascape database and Starbase database were used to analyze the functional enrichment of disease-syndrome efficacy-related modules at two time points,and the pathways and functions of related genes in the two time-point modules were compared.The LncBase Predicted database was used to predict the correlation between lncRNA and miRNA;Targetscan,miRDB,miRTarBase,and miRcode databases predicted the correlation between miRNA and mRNA to construct a ceRNA network,conduct topology analysis on the network to mine core nodes,and analyze the regulation in the network.To explore the relationship and analyze the molecular mechanism of Danhong injection in the treatment of chronic stable angina pectoris with blood stasis syndrome.The miEAA database and LncSEA database were used to enrich miRNA and lncRNA at two time points,and the Metascape database(the parameter was set to Min Overlap=1,P<0.05,Min Enrichment=1)was used to enrich mRNA,and the Starbase database supplemented enrichment set analysis.4 Results1.On the 14th day,lncRNA had 56 differential modules and 14456 differential lncRNAs;miRNA had 29 differential modules and 364 differential miRNAs;mRNA had 43 differential modules and 2208 differential miRNAs.On the 14th day,the disease-syndrome efficacy-related modules were lncRNA paleturquoise module,miRNA midnightblue module,and mRNA darkred module as disease-syndrome efficacy-related modules.There are 14 miRNAs,1 newly discovered miRNA,35 mRNAs,and 83 lncRNAs in the disease-syndrome-efficacy-related modules.The 14-day ceRNA network was constructed by analyzing the interaction relationship between the modules related to disease,syndrome and efficacy through the database.There were 13 miRNAs,11 mRNAs,and 7 lncRNAs in the network,with a total of 38 edges.2.On the 30th day,lncRNA had 67 differential modules and 14467 differential lncRNAs;miRNA had 26 differential modules and 297 differential miRNAs;mRNA had 79 differential modules and 1895 differential miRNAs.On the 30th day,disease-syndrome efficacy related modules lncRNA orange module,miRNA salmon module,and mRNA darkturquoise module.There are 16 miRNAs,3 newly discovered miRNAs,46 mRNAs,and 85 lncRNAs in the 30-day disease-syndrome efficacy-related modules.In the 30-day network,7 miRNAs,13 mRNAs,and 2 lncRNAs had interaction relationships,with a total of 23 edges.3.Using the miEAA database and the LncSEA database to enrich miRNA and lncRNA at two time points,the pathway was not enriched.Using Metascape database and Starbase database analysis,it was found that 104 biological processes were involved in GO enrichment on the 14th day,such as mammary duct morphogenesis,positive regulation of epidermal growth factor-activated receptor activity,histone regulation,triglyceride transport,coagulation regulation and other processes 32 cellular components,related to nuclear RNA export factor complexes,platelet granules,lipoprotein granules,etc.;31 molecular functions,binding to epidermal growth factor receptor,hyaluronan glucosaminidase activity,hexosaminidase activity,lipoprotein lipase activator activity,etc.KEGG pathway enrichment analysis showed that a total of 55 pathways were enriched,including Erbb signaling pathway,Ppar signaling pathway,drug metabolism cytochrome P450 pathway,glutathione metabolism pathway,Wnt signaling pathway,P53 signaling pathway,TGFβ signaling pathway,MAPK signaling pathway,Jak Stat signaling pathway,Mtor signaling pathway,etc.On the 30th day,GO enrichment analysis of disease-syndrome-efficacy-related modules revealed that 117 biological processes were involved,such as SRP-dependent co-translational protein targeting membrane signal sequence recognition,protein targeting,regulation of mitotic cell cycle transition,nerve impulse conduction,neural Transmitter regulation,etc.;17 cellular components,such as Schwann cell microvilli,smooth endoplasmic reticulum,filamentous actin,etc.;43 molecular functions,such as potassium channel activity,cation channel activity,steroid hormone receptor activity,cholesterol dehydrogenase activity,etc.KEGG was enriched to 52 pathways,involving Wnt signaling pathway,Erbb signaling pathway,MAPK signaling pathway,Gnrh signaling pathway,KEGG P53 signaling pathway,Tgf Beta signaling pathway,steroid biosynthesis,glycolysis gluconeogenesis,viral myocarditis,pyruvate metabolism,insulin signaling pathway,apoptosis and other signaling pathways.4.The network on the 14th day and the network on the 30th day have the same network diameter,indicating that the two networks have the same ability to deal with internal and external interference factors.The network characteristic path length,average degree centrality,network density,and network heterogeneity between the two time points were less different.The connected subgraph of the network on the 30th day was lower,and the network connectivity might be stronger.Through the betweenness centrality value of the network,it is found that the key node in the 14-day network is hsa-miR-516b-5p,and its betweenness centrality value is 0.40;the key node in the 30-day network is hsa-miR-6796-3p,whose mediation The number centrality value is 0.51.According to the key nodes,ENST00000549163.1-hsa-miR-516b-5p—CSMD2/GABRA3/MARK1/SCN1A/SLC44A5 in the network on the 14th day,ENST00000434195.1—hsa-miR-6796-3p—ACSM5 in the network on the 30th day GALR1/NFASC/ST8SIA5 may have a regulatory relationship,which has a key impact on the network.Through analysis and verification,it was found that hsa-miR-6796-3p-NFASC has a common pathway,which may have a regulatory relationship.5 Conclusions1.The disease-syndrome efficacy-related modules of Danhong injection in the treatment of chronic stable angina pectoris and blood stasis syndrome at the 14th day and the 30th day were determined by the weighted gene co-expression network analysis method,and the ceRNA network was constructed based on the interaction relationship..2.The disease-syndrome efficacy-related modules on the 14th day and the disease-syndrome efficacy-related modules on the 30th day both act on Erbb pathway,Wnt pathway,P53 pathway,TGFβ pathway,MAPK pathway,insulin signaling pathway,phosphatidylinositol signaling system,and B cells.Receptor signaling pathway,the drug metabolism cytochrome P450 signaling pathway was enriched in the module on day 14,and there were more glucose and lipid metabolism-related pathways enriched in the module on day 30.The key nodes in the ceRNA network on the 14th and 30th days are hsa-miR-516b-5p(betweenness centrality value 0.40)and hsa-miR-6796-3p(betweenness centrality value 0.51).have a potential regulatory relationship.3.The study found that the pathological factors of angina pectoris,the manifestations of blood stasis syndrome,the correlation module analysis of Danhong injection treatment mechanism and the disease syndrome efficacy,and the nodes in the ceRNA network all involve Erbb pathway,Wnt pathway,P53 pathway,TGFβ pathway,MAPK pathway,and insulin signaling The study suggests a way to explore the mechanism of traditional Chinese medicine compound treatment of diseases.