Network Biological Mechanisms For Phlegm-Stasis Cementation Syndrome With Ischemic Heart Disease

ObjectTo explore network biological mechanisms for phlegm-stasis cementation syndrome with ischemic heart disease.Materials and MethodsIn the first step,ischemic heart disease(IHD)-related genes was obtained by integrating the CADgene database,the Pubmed literature,the Online Mendelian Inheritance in Man database(OMIM),and the GWAS database(Genome-Wide Association Study,GWAS)In the second step,IHD-related genes was mapped into the STRING V9.1 protein-protein interaction network(PPI),and the molecular network modules related to IHD(disease module)were determined based on the enrichment results.In the third step,the symptom-gene relationship of phlegm-stasis cementation syndrome with ischemic heart disease(PSCS)obtained from the Human Symptoms-Disease Network(HSDN)was projected to the disease modules,the modules relevant to PSCS(Syndrome Module)were determined according to the enrichment situation.Molecular Network Module.A preliminary verification of the syndrome modules was performed from three aspects:1)Known classical formulas-drug-active-target relationships from Herb Ingredients’ Targets(HIT).Danshen Decoction Taohong Siwu Decoction,Gualou Baibanxia Decoction and Huanglian Wendan Decoction were selected as classical formulas.2)Known drug-target relationship for IHD from Therapeutic Target Database(TTD)and DrugBank database.3)Phenotype-gene relationship from Phenotype Ontology(HPO)In the fourth step,the genes relevant to PSCS repored in the literature review were projected to the syndrome module,and the position of these genes in the syndrome module in the literature were calculated.The genes reported in the literature review and their First Neighbors,the nearest neighbor node constituted the relevant molecular network of PSCS.In the fifth step,a cross-sectional,control,small sample,and paired study designs using clinical sample expression profiling(RNA sequencing)was conducted group comparison analysis and correlation analysis were performed simultaneously to obtain PSCS-related genesIn the sixth step,the resutls from the literature reports,data mining and clinical research was support each other;and the related pathways associated with the genes involved in data mining and clinical research were closely related to PSCSResults(1)A total of 1,056 IHD-related genes was obtained.(2)A total of 8 disease modules were obtained(Module 195,204,95,203,194,212,59,146).(3)A total of 2 syndrome modules(Module195,95)were obtained.Preliminary verification results show that a total of 19 formulas(blood stasis)-herbs-drugs-active ingredients-targets,15 formulas(phlegm)-drug-effective ingredients-targets,7 drug-targets for IHD,and 3 pairs of symptoms-genes relationship appeared in Module 95.a total of 3 formulas(blood stasis)-herbs-drugs-active ingredients-targets,1 formulas(phlegm)-drug-effective ingredients-targets,4 drug-targets for IHD,and no symptoms-genes relationship appeared in Module 195.(4)A total of two molecular networks closely related to PSCS were obtained.Among them,Module 95 contains 8 related genes reported in the literature review,and these 8 genes and their first neighours constituted PSCS relevant molecular network A.The gene and its first neighours constituted PSCS relevant molecular network B(5)In the Cross-sectional,small-sample,matched-paired,controlled clinical study.a total of 9 healthy patients(control group)and 21 IHD patients(disease groups)were recruited.After group comparison analysis and correlation analysis,a total of 118 PSCS-associated genes were obtained.(6)The results of data mining and clinical studies showed that the relevant pathway for PSCS was Leukocyte Extravasation Signaling,and TIMP-3,MMP-8 and MMP-9 may play a key role.ConclusionThrough data mining and clinical validation,it was found thatPSCS was closely related to the Leukocyte Extravasation Signaling pathway,among which MMP-8,MMP-9 and TIMP-3 may play a key role.Further analysis of its function indicated that PSCS was closely related to the inflammatory response.