| Objective: To investigate the expression and clinical significance of plasma Exosomal mi RNA in heart failure(HF)patients.Methods: 92 HF patients and 52 healthy controls were enrolled for our study,the clinical date of those objects were collected for analysis.There were 23 NYHA II,40 NYHA III,29 NYHA IV patients according to NYHA classification and45 ischemic heart disease,32 dilated cardiomyopathy,15 valvular heart disease when classified by etiology.Exosomes were isolated from plasma by special reagent,transmission electron microscope(TEM),laser particle size analysis and exosome surface markers(CD63,CD81)detection were preformed to identification.Ten kinds of mi RNAs including mi R-1,mi R-21,mi R-22,mi R-23 a,mi R-24,mi R-29 a,mi R-133,mi R-210,mi R-214,mi R-423-5p which was closely related to HF and(or)ventricular remodeling were selected for our study.We quantified the relative expression level(2-??Ct)of these Exosomal mi RNAs in 12 HF patients and 12 healthy subjects with RT-PCR.Exosomal mi RNAs which were differentially expressed were selected in order to further analyze.Furthermore,we analyzed the relationship between mi RNA relative expression level and part of clinical date.ROC curve was used to evaluate the application of Exosomal mi RNA in the diagnosis of HF.Results:1.Plasma exosomes were analysed under TEM and grainsize analyzer which showed that the nanoparticles have a generally spherical shape with diameters mainly range from 40 nm to 100 nm.Surface marker detection verified that CD63 and CD81 was positive on exosome.2.The screening results of 10 selected mi RNAs show : relative expression level of plasma Exosomal mi R-21?mi R-210?mi R-423-5p were higher in HF patients(P<0.05),the relative expression level was 2.07± 0.77,3.49± 2.93,6.16 ±5.70;while Exosomal mi R-133 was lower(P<0.05),the relative expression level was0.58 ± 0.19.Further validation study showed that only high expression level of Exosomal mi R-210 and mi R-423-5p had its statistical significance(P<0.05),the relative expression level was 3.39±2.01,6.39±4.15.while the relative expression level of Exosomal mi R-133 and mi R-21 was 0.89±0.50?1.22±0.46,it showed no significant differently between HF patients and healthy controls(P>0.05).3.Plasma level of Exosomal mi R-210 and mi R-423-5p were increase along with the increase of NYHA classification.The relative expression level of Exosomal mi R-210 in NYHA II,NYHA III and NYHA IV groups was 2.51±1.38?2.99±2.13?4.74±2.08,the relative expression level between NYHA III and IV,NYHA II and IV showed significant difference(P < 0.05).Exosomal and mi R-423-5p had significant difference among three NYHA groups(P < 0.05),the relative expression level was 3.87±3.46? 6.04±3.90?6.04±3.90.It showed no statistical significance of plasma level of Exosomal mi R-210 and mi R-423-5p among ischemic heart disease,dilated cardiomyopathy and valvular heart disease(P >0.05).The relative expression level of Exosomal mi R-210 and mi R-423-5p were positively correlated with log2(NT-pro BNP)(r=0.53,P<0.05;r=0.63,P<0.05)and were negative correlated with LVEF(r=-0.46,P<0.05;r=-0.51,P<0.05),but there's no correlation with LVEDd(r=0.19,P=0.07;r=0.17,P=0.11).4.Area Under roc Curve for Exosomal mi R-210 and Exosomal mi R-423-5p were0.79 and 0.91.The AUC would reach 0.93 when combine these two target when the cut-off points were 0.71,the seneitivity was 88.46% and specificity was 82.61%Conclusion: The plasma Exosomal mi RNA are significantly dysregulated in HF.Exosomal mi R-210 and Exosomal mi R-423-5p may have potential clinical application value in field of diagnosis and evaluation of HF. |
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