Proteomics Study Of Heart Yang Deficiency Syndrome In Chronic Heart Failure And Screening Of Potential Marker

Objective:1.To study the differential protein of heart yang deficiency syndrome in chronic heart failure,reveal its microscopic material basis,and explore the biological basis of the disease.2.Screening potential biomarkers of heart yang deficiency syndrome in chronic heart failure and constructing diagnostic models to provide a basis for further study of pathogenesis and accurate syndrome differentiation.Methods:1.In this study,a total of 24 patients with chronic heart failure were included as the test set,including 12 patients with heart yang deficiency syndrome,12 patients with heart non-yang deficiency syndrome,and 12 healthy controls during the same period.The three groups had homogeneity in gender,age,course of disease,concomitant disease and grading of cardiac function.DIA proteomics method was used for protein analysis of serum samples from the three groups.The data after mass spectrometry analysis was compared with Uniprot database to obtain protein identification results.According to Fold Change ≥ 2.0 and P < 0.05,the differently-expressed proteins between groups were obtained.Protein interaction,GO and KEGG Pathway were used to analyze bioinformatics of differentially expressed proteins.2.In this study,76 patients with chronic heart failure were included as a validation set,which did not duplicate the test set.Among them,35 cases were heart yang deficiency syndrome and 41 cases were heart non-yang deficiency syndrome.Clinical indicators of patients in the two groups were collected,and the concentrations of serum differential proteins(MBL,ADP,IGFBP2,IGFBP4)of patients with heart yang deficiency syndrome and heart non-yang deficiency syndrome were detected by ELISA.ROC curve analysis of clinical indicators and differential proteins was performed to calculate the area under the curve and determine the diagnostic value.The optimal diagnostic model was established by logistic regression analysis,and the accuracy,sensitivity and specificity of the diagnostic model were analyzed by ROC curve.Results:1.Proteomic study of heart yang deficiency syndrome in chronic heart failure: a total of 139 differential proteins were screened from heart yang deficiency syndrome group compared with healthy control group,including80 up-regulated proteins and 59 down-regulated proteins.A total of 65 differential proteins were screened from the heart non-yang deficiency syndrome group compared with the healthy control group,including 52up-regulated proteins and 13 down-regulated proteins.By protein interaction analysis,common core differential proteins included C-reactive protein(CRP),serum amyloid A1(SAA1),insulin-like growth factor-1(IGF1),βactin(ACTB),etc.However,some disease-related proteins,such as adiponectin(ADP),insulin-like growth factor binding protein 2(IGFBP2),insulin-like growth factor binding protein 4(IGFBP4),were only found in CHF heart yang deficiency syndrome.GO and KEGG Pathway enrichment analysis was conducted for the differential proteins among each group.KEGG Pathway was enriched in immune system and other signaling pathways in both groups,while glycerophospholipid metabolic Pathway was only enriched in heart yang deficiency syndrome of CHF,suggesting that the pathology of CHF is closely related to immune response.Abnormal disturbance of glycerol phospholipid metabolic pathway appeared in heart yang deficiency syndrome.Antimicrobial humoral response(antimicrobial humoral response),defense response to bacterium(defense response to bacteria)and other biological processes are highly enriched in the differential proteins of heart non-yang deficiency syndrome.These results indicate that heart non-yang deficiency syndrome in CHF is regulated in these processes.A total of 45 different proteins were screened out in chronic heart failure syndrome with heart yang deficiency compared with heart non-yang deficiency syndrome.The protein interaction analysis showed that 17 of them were up-regulated,including HP(haptoglobulin),ADP(adiponectin),Apo E(apolipoprotein E),SPP1(Secreted Phosphoprotein 1),etc.The number of down-regulated proteins was 28,mainly including MBL(mannose-binding agglutinin)and HRG(histidine-rich glycoprotein).These results suggest that these proteins may reveal the microscopic substance changes in the macroscopic syndromes of heart yang deficiency syndrome in chronic heart failure.2.Screening of potential markers of heart yang deficiency syndrome in chronic heart failure: compared with heart non-yang deficiency syndrome in chronic heart failure,the concentration of platelet count,ion calcium and apolipoprotein A decreased P<0.05),and the concentration of urea nitrogen,creatinine and myoglobin increased(P< 0.05);After ROC curve analysis:The area under platelet count curve was 0.682,ion calcium curve was 0.649,apolipoprotein A curve was 0.783,urea nitrogen curve was 0.637,creatinine curve was 0.662,myoglobin curve was 0.644,all of which had statistical significance(P<0.05).Proteomic results verified by ELISA showed significant differences in MBL,ADP,IGFBP2 and IGFBP4 between xinyang deficiency syndrome and heart non-yang deficiency syndrome,which were consistent with proteomic results.After ROC curve analysis: taking heart yang deficiency syndrome as the target,the area under MBL curve was 0.673,the best predictive value was 1438.39ng/ m L,and the Jordon index was 0.524.The area under the ADP curve was 0.699,the best predictive value was 45951.97ng/ m L,and the Yuden index was 0.524.The area under the IGFBP2 curve was 0.902,the best predictive value was 618.26ng/ m L,and the Yuden index was 0.715.The area under the curve of IGFBP4 was 0.908,the best predictive value was 63.3ng/ m L,and the Yuden index was 0.732.The results showed that the four differential proteins had certain diagnostic value for xinyang deficiency syndrome,and had statistical significance(P < 0.05).Binary Logistic regression analysis showed that the diagnostic model composed of apolipoprotein A,IGFBP2 and IGFBP4(Logistic regression equation Logit(P)=(2.234+3.546 × Apolipoprotein A-0.009 × IGFBP2-0.018 ×IGFBP4)was the optimal combination.ROC curve analysis showed that the area under curve value,sensitivity and specificity were 0.962,97.1% and90.6%,respectively,with statistical significance(P<0.05).Conclusion:1.There are differentially expressed proteins in serum of chronic heart failure patients with heart yang deficiency syndrome and patients with heart non-yang deficiency syndrome.The differential proteins mainly included MBL,ADP,IGFBP2,IGFBP4,HP,Apo E,SPP1,HRG,etc.,which preliminarily explained the biological basis of the disease.The central differential protein of heart yang deficiency syndrome is involved in glycerol phospholipid metabolism and other signaling pathways,which explains the biological nature of heart yang deficiency syndrome in chronic heart failure from the perspective of biomolecular network.2.Platelet count,urea nitrogen,creatinine,myoglobin,apolipoprotein A,calcium,MBL,ADP,IGFBP2,IGFBP4 and other 10 indicators are of significance for the syndrome type diagnosis of heart yang deficiency syndrome in chronic heart failure to A certain extent.The combined diagnostic model of apolipoprotein A,IGFBP2 and IGFBP4 was preliminarily identified as the optimal combination,which could be used as A potential biomarker for heart yang deficiency syndrome of chronic heart failure and provide guidance for clinical diagnosis.