Analysis Of ADAMTS2 Expression Difference In Serum Of Patients With Dilated Cardiomyopathy Based On Transcriptome Sequencing

Objectives:1.To analyze the differential expression of mRNA in peripheral venous blood of patients with Dilated cardiomyopathy(DCM)and the control group,to screen the differential genes of dilated cardiomyopathy(DCM),and to find the specific molecular markers of DCM.2.Validation of ADAMTS2 protein expression in sera from patients with dilated cardiomyopathy and controls to provide a research basis for early diagnosis of DCM.Methods:1.A case-control study was conducted.Three patients diagnosed with DCM in the First Affiliated Hospital of Kunming Medical University from September 2020 to June 2022 were selected as the case group,and three healthy volunteers who underwent physical examination in the First Affiliated Hospital of Kunming Medical University during the same period were selected as the control group.Peripheral blood mononuclear cells(PBMCs)were isolated from the peripheral venous blood of the two groups by centrifugation.After RNA extraction and quality control,RNA was pretreated to construct a sequencing library.RNA sequencing was performed on the Illumina HiSeq sequencer,and the raw data were obtained and quality control was performed on Q30.The Hisat tool was then used to align the high-quality,unspliced Valid Data with the reference genome.Then the FPKM value of gene level mRNA was obtained by cuffdiff software,and the p-value and fold change between the two groups of samples were calculated to obtain the differentially expressed mRNA expression profile.The Kyoto Encyclopedia of Genes and Genomes(KEGG)and DAVID(Database for Annotation,GO)databases were applied.Visualization and Integrated Discovery(IDP)online tools were used to study the biological function of the differential gene expression profile of DCM,and to improve the disease association analysis.Then STRING database was used to construct protein-protein interaction(PPI)network of differential genes,and cytohubba was used to screen out the key genes related to DCM.2.Thirty patients admitted to the First Affiliated Hospital of Kunming Medical University and diagnosed with DCM from September 2020 to June 2022 were selected as the case group,and 28 healthy volunteers who underwent physical examination in the First Affiliated Hospital of Kunming Medical University during the same period were selected as the case control group.Whole peripheral venous blood of the two groups was extracted and centrifuged to obtain serum.ADAMTS2 protein was detected by ELISA.ADAMTS2 protein content in the experimental results of the two groups was analyzed for statistical analysis.Whole peripheral venous blood of the two groups was extracted and centrifuged to obtain serum.ADAMTS2 content in serum of DCM group and control group was detected by Enzyme-linked immunosorbent assay(ELISA),followed by statistical analysis.3.Basic information and clinical data were collected for each subject,including:age,sex,history of hypertension,history of smoking,history of alcohol consumption,whether other diseases were combined,clinically relevant biochemical parameters(fasting glucose,total cholesterol,LDL,HDL,triglycerides,BNP),echocardiographic data(left atrial internal diameter,left ventricular end-diastolic internal diameter,right atrial length,LVEF,LVFS,LVEDV,LVESV,SV,CO,CI,ascending aortic internal diameter),New York Classification of Cardiac Function(NYHA),and statistical analysis.LVFS,LVEDV,LVESV,SV,CO,CI,ascending aortic internal diameter),New York Classification of Cardiac Function(NYHA),and statistical analysis were performed.Results:1.Transcriptome sequencing prompted results suggesting differential gene expression between patients with dilated cardiomyopathy and normal controls,with1843 differentially expressed genes,of which 664 were up-regulated and 1179down-regulated;a total of 2600 differentially expressed transcripts,of which 1444 were up-regulated and 1156 were down-regulated.2.A total of 387 significantly differentially expressed genes were screened out from 1843 differentially expressed genes obtained by transcriptome sequencing,and then the significantly differentially expressed genes were used to construct PPI network.At the same time,387 differential genes were combined with 4706disease-related genes of DCM in Gene Cards,and 113 key genes were obtained after intersection.The key gene-protein interaction network of dilated cardiomyopathy was preliminarily constructed,and 15 hub genes were screened by Cytoscape.ADAMTS2 may be one of the pathogenic genes of dilated cardiomyopathy.3.The age,left atrial diameter,left ventricular end-diastolic diameter,right atrial length diameter,LVEDV,LVESV,HS-CRP,PCT,AST,creatinine,uric acid,BNP levels and HYHA classification in the DCM group were significantly higher than those in the normal control group,and the probability of heart failure,pulmonary hypertension and arrhythmia were significantly higher than those in the normal group.All the above differences were statistically significant(P < 0.05).There were statistically significant differences in gender,ALT,serum potassium level,serum sodium level and ascending aorta diameter between the two groups(P < 0.05).4.HDL-C,LVEF,LVFS,exercise amplitude,CO,and CI were significantly higher in the normal control group than in the DCM group,and the differences were all statistically significant(P< 0.01).5.Serum ADAMTS2 level in the normal control group was significantly lower than that in the DCM group,and the difference was statistically significant(P <0.001).6.The level of serum ADAMTS2 is a risk factor for the development of DCM.7.Right atrial internal diameter and LVESV were independent risk factors for the development of heart failure in patients with DCM.8.The combined serum levels of ADAMTS2,left atrial internal diameter,left ventricular end-diastolic internal diameter,right atrial long diameter,LVEF,LVFS,LVEDV,LVESV are multifactorial factors with high predictive value for the development of heart failure in patients with DCM.Conclusions:1.The elevated serum ADAMTS2 level may be one of the risk factors for DCM.The preliminary sensitivity and specificity were 0.933 and 0.964 respectively.2.The serum ADAMTS2 level combined with left atrial diameter,left ventricular end-diastolic diameter,right atrial length diameter,LVEF,LVFS,LVEDV and LVESV have high predictive value for the occurrence of heart failure in DCM patients.