| Objective Urinary biomarkers for the diagnosis of chronic heart failure（CHF）were screened based on 4D-LFQ proteomics technology,and CTSA and DDP4 levels were further measured to explore their clinical diagnostic value for CHF patients.MethodsAll proteins in urine of CHF group（n=10）and NC group（n=10）were analyzed qualitatively and quantitatively by 4D-LFQ proteomics technique,and screening for significantly differentially expressed proteins.Bioinformatics software was used to analyze the functional annotation and enrichment analysis of differentially expressed proteins and the interaction between different proteins.The levels of CTSA,FUCA,AGT,DDP4 and GLB1 in urine of CHF group（n=48）and NC group（n=40）were quantitatively measured by enzyme-linked immunosorbent assay（ELISA）.The diagnostic value of urinary CTSA,FUCA and DDP4 in chronic heart failure was evaluated by receiver operating characteristic（ROC）curve analysis.Pearson correlation analysis was used to explore the correlation between urinary CTSA and DDP4 expression levels and LVEF,LAD,LVESD,LVEDD,NT-pro BNP,Scr and e GFR.Results（1）A total of 2047 proteins were identified by mass spectrometry and protein identification in our study,of which 1477 proteins could be quantitatively analyzed.We found that 82 urinary proteins,including CTSA,DDP4,FUCA,AGT and GLB1,were significantly upregulated and 118 were downregulated based on the criteria of multiple of difference >1.5 and<1/1.5.（2）Urine samples were collected from 48 CHF patients and 40 normal controls.ELISA showed that compared with NC group,urinary CTSA level [2551.4（2084.76-2785.9）vs 3055.43（2498.33-3434.76）pg/m L,P<0.0001],DDP4 level [254.46（161.74-329.02）vs 2082.51（667.0-2418.6）ng/m L,P<0.0001] and FUCA level [151.92（64.98-447.64）vs 326.42（139.46-639.62）μU/m L,P=0.05] were significantly increased in CHF group,and the differences were statistically significant.There were no significant differences in the levels of urine AGT and GLB1 between the two groups（P all> 0.05）.（3）ROC curve analysis showed that urinary CTSA and DDP4 could differentiate chronic heart failure patients from normal controls（AUC was 0.732,0.875,sensitivity was 64.6%,79.2%,specificity was 75%,92.5,respectively）,while the diagnostic value of urine FUCA was lower（AUC=0.642,Sensitivity was 81.3% and specificity was 45.8%）.（4）Pearson phase analysis showed that the level of urinary CTSA was negatively correlated with LVEF（r=-0.448,P<0.0001）,was positively correlated with LAD,LVESD,LVEDD and NT-pro BNP levels（r=0.317,r=0.38,r=0.308,r=0.279,P all<0.01）,was not associated with Scr and e GFR levels（P all>0.05）;Urinary DDP4 level was positively correlated with LVEF（r=0.489,P<0.0001）,was negatively correlated with LAD,LVESD,LVEDD,NT-Pro BNP levels and e GFR（r=-0.395,r=-0.512,r=-0.492,r=-0.274,r=-0.293,P all<0.05）and was not associated with Scr levels（P>0.05）.Conclusions（1）Urinary proteomics based on 4D-LFQ technique is a reliable method to find biomarkers for disease diagnosis.（2）The concentration of urinary CTSA and DDP4 were increased in patients with chronic heart failure.The level of urinary CTSA was positively correlated with LAD,LVESD,LVEDD and NT-pro BNP,and negatively correlated with LVEF.The level of urinary DDP4 was negatively correlated with LAD,LVESD,LVEDD and NT-pro BNP,and positively correlated with LVEF.（3）Urinary CTSA and DDP4 can be used as potential biomarkers for the diagnosis of chronic heart failure. |
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