The Transcriptomics Study Of Different Therapeutic Effect Of Xinyin Tablets In Treating Coronary Heart Disease Chronic Heart Failure(Qi Yin Deficiency,blood Stasis And Water Retention Syndrome)

Objective:Xinyin tablets have significant clinical effects on patients with chronic heart failure(Qi Yin deficiency,blood stasis and water retention syndrome).It can improve cardiac systolic function,improve NYHA cardiac function classification,exercise endurance,and reduce the incidence of ventricular arrhythmia.In this study,peripheral leukocyte RNA-seq technology was used to analyze the changes of patients’ white blood cells in different groups before and after taking Xinyin Tablets.Exploring the possible mechanism of Xinyin Tablets by analyzing the differential gene expression and related biological processes and signal pathways between the excellent and effective groups.Methods:This study was reviewed by the Medical Ethics Committee,and all subjects signed an informed consent form.A total of 11 patients with coronary heart disease and chronic heart failure in the Cardiovascular Outpatient and Inpatient Department of the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine were included in accordance with the criteria for inclusion.On the basis of standard western medicine treatment,supplemented with Chinese patent medicine Xinyin tablets(specification: 0.25g/tablet),4 tablets/time,3 times/day,with warm boiled water,and the course of treatment is4 weeks.The basic information of the patient is recorded in detail,the efficacy indicators and safety observation indicators(hematuria routine,electrocardiogram,liver and kidney function,etc.)are evaluated at the two visit points before and after treatment,and then the patients are divided into categories The excellent group and the effective group are used as transcriptome research groups.At the same time,4ml of peripheral blood was drawn for white blood cell extraction at the two visit points before and after treatment,and 1ml of trizol reagent was added and stored in the refrigerator at-80°C.After all samples were collected,the reference transcriptome sequencing analysis was performed.The transcriptome differences between the excellent and effective groups before and after taking Xinyin Tablets were compared.We are going to explore the candidate genes and signal pathways of Xinyin Tablets for the treatment of coronary heart disease and chronic heart failure through the differentially expressed genes(DEGs)analysis,Gene Ontology(GO)functional annotation and Kyoto Gene and Gene Database(Kyoto Encyclopedia of Genes and Genomes,KEGG)enrichment analysis process.Results:1.Clinical observationThe patients were divided into the excellent group and the effective group according to the criterion of efficacy by analyzing the examination results before and after the patients were enrolled.There were 3 males and 2 females in the excellent group,4 patients whose cardiac function classification improved by 1level according to the NYHA cardiac function classification and 1 patient reached Class I(P=0.025<0.05).The result was statistically significant.There were 5 patients in the effective group,including 2 males and 3 females.The NYHA cardiac function classification of all patients did not change.2.Transcriptomics researchPeripheral blood leukocyte RNA-seq was used to analyze the difference in gene expression between the excellent group and the effective group before and after taking Xinyin Tablets.The P value <0.05 and fold change>1.5 were set as the screening criteria for differentially expressed genes.Then we conducted a preliminary research on the main biological processes,functions and signal transduction pathways that the all DEGs participated in by analyzing the GO and KEGG pathway analysis.A total of 129 DEGs were screened from pro＿eff-vs-pre＿eff,of which 33 were up-regulated differentially expressed genes(including EXOC3L2,ANGPTL4,STC2,etc.)and 96 were down-regulated differentially expressed genes(including IFI6,COL6A3,PMAIP1,IDO1,CXCL8,etc.).There were 105 DEGs were screened from pro＿exc-vs-pre＿exc,including 41 up-regulated differentially expressed genes(including FN1,LOC118,SPARCL1,etc.)and down-regulated 64 differentially expressed genes(including COL12A1,CXCL8,SPOCK1,COL6A3,CAMP,KIR3DL1,etc.).GO analysis showed that the DEGs of pro＿eff-vs-pre＿eff were enriched in cell adhesion,signal transduction,protein extracellular matrix,nucleus,type I interferon signaling pathway,and negative regulation of cell proliferation.While the DEGs of pro＿exc-vs-pre＿exc were enriched in cell adhesion,signal transduction,protein extracellular matrix,cell nucleus,collagen fibril organization、collagen catabolic process、negative regulation of cell proliferation、apoptotic process.KEGG analysis found that the two groups of DEGs were mainly enriched in the antigen presentation,natural killer cell-mediated cytotoxicity,Toll receptor signaling pathway,tumor necrosis factor signaling pathway,interleukin 17 signaling pathway,cytokine-cytokine receptor interaction and PI3K-Akt The signal pathway.Those were closely related to the body’s immune inflammatory response.Conclusion:In this study,we divided patients into excellent and effective groups according to the different therapeutic effect of Xinyin tablets.Based on the analysis of transcriptomics,Xinyin Tablets may exert efficacy through the following two aspects:(1)The down-regulation of COL12A1,ADAMTS2,COL5A2,etc.may inhibit immune inflammatory response and regulate the structure,metabolism,and expression of collagen fibrin.The the process of myocardial fibrosis may be inhibited by the down-regulating genes and clinical symptoms be alleviated.(2)PI3K-Akt signaling pathway,Cytokine-cytokine receptor interaction and other signaling pathways,regulated by the down-regulation of CXCL8,CXCL1,FN1,COL1A2 and other genes,may inhibit myocardial fibrosis by inhibiting immune inflammation and down-regulating autophagy.