Differences Of MiRNAs Expression In Acute Stanford A/B Aortic Dissection And Their Significance

Background and objectiveAcute Aortic Dissection(AAD): It is a critically important vascular disease,mainly caused by various causes of arterial cystic changes with elastic fibers rupture,necrosis,resulting in defects in the vessel wall structure,blood within the rupture Membrane entry into the middle layer led to delamination of the wall,forming a double-cavity aorta,depending on whether the rupture of the endocardium involved the ascending aorta into the Stanford A and B type.If the A type AAD is not treated in time within 48 hours,the mortality rate is as high as half,and the condition is extremely dangerous.If the early diagnosis and classification can be accurately performed,a reasonable choice of treatment plan can effectively improve the prognosis.At present,AAD relies mainly on imaging studies to confirm diagnosis.However,due to its time-consuming,expensive operation and the need to repeatedly move patients,emergency conditions and basic-level hospitals are sometimes unable to perform,and it is easy to delay the optimal timing of treatment.Initial screening indicators.MicroRNA(miRNA)is a non-coding small regulatory nucleotide that is stably present in the peripheral blood.It is an important transcription factor and post-transcriptional inhibitor of gene expression.It participates in the pathogenesis of various diseases and can be detected in peripheral tissues.The level varies with the evolution of the disease.Studies have shown that miRNAs are also involved in the pathogenesis of aortic dissection.The miRNAs-4787-5P and miRNA-4306 previously screened in this study have high sensitivity and specificity for the early diagnosis of acute aortic dissection and may become early diagnosis.The potential biomarkers,and predicted and confirmed their target genes are PKD1 and TGF?1,respectively,many studies have shown that they are related to the pathogenesis of aortic dissection,it is speculated that the two miRNAs may also be involved in the incidence of aortic dissection mechanism.In this study,based on previous studies,we further screened different miRNAs with different expression levels between two different types of AAD,and explored whether miRNAs have a certain diagnostic value for the classification of AAD,and analyzed the differences and significance.Research methodsThe peripheral blood plasmas of the first affiliated hospital of Zhengzhou University and were diagnosed as AAD patients and normal controls were collected from October 2016 to November 2017,respectively(all the participants had signed the informed consent after being approved by the Hospital Ethics Committee).Detailed records of all vital signs(blood pressure,heart rate,respiration)and blood plasma(WBC),platelet(PLT),D-dimer,and C-reactive protein within 24 hours of admission to all enrollees(C-reactive protein,CRP),atherosclerosis,lipid profile,brain natriuretic peptide(BNP),ascending aorta width and other basic clinical data.The enrollees were divided into three groups: Group A(acute Stanford A)and Group B(acute Stanford B)and Group C(normal control).Six cases from three groups of plasma samples were selected to extract total RNA in the blood samples for quality testing.The expression profiles of miRNAs were analyzed using Agilent miRNA microarrays,and three groups of miRNAs with significant differences were screened out.Fluorescence quantitative PCR was applied to miRNAs(miRNA-146 b,miRNA-29 b,and miRNA-21)with significant differences in primary screening results were validated in three groups of samples,and the three differentially expressed miRNAs were analyzed using receiver operating characteristics(ROC).The specificity and sensitivity of the early diagnosis of AAD,and finally through the analysis of bioinformatics technology to predict the downstream target genes of miRNA-146 b,miRNA-29 b and miRNA-21,to analyze the acute Stanford A and B type aortic dissection Differential expression and significance of miRNAs.Results1.A total of 115 patients with acute aortic dissection were included(51 with Stanford A and 64 with B),of which 10 were female A and 41 were male;13 were B female,51 were male;48 were normal controls.There are 39 males and 9 females.There was no significant difference in gender,age,history of hyperlipidemia,drinking history,smoking history,and RBC count between groups A,B,and C(all P>0.05).Atherosclerosis and hypertension were found in groups A and B.Cases were significantly higher than those in group C,and cases with combined type B and atherosclerosis were higher than those in group A and C.The history of type A dissection combined with hypertension was significantly higher than that of group B and C,and the differences were statistically significant(P <0.05);A,B group ascending aorta width was significantly wider than the C group,and A group ascending aorta width was wider than the B group,the difference was statistically significant(P <0.05);The D-dimer values,BNP values,WBC counts in group A and B were significantly higher than those in group C,and the three groups in group A were also significantly higher than those in group B(P<0.05).The CRP levels in the two groups were significantly higher than those in the C group(P<0.05).There was no significant difference in CRP between the two groups.The CK-MB in the A group was significantly higher than that in the B and C groups.The difference was statistically significant(P<0.05),but there was no significant difference in CK-MB between groups B and C;HB in group A and B was lower than that in group C,and A group was significantly lower than that in group B,and the differences were statistically significant.Significance(P<0.05);A,B group PLT decreased compared with C group,the difference was statistically significant(P<0.05),A,B between the two groups PLT no significant difference.2.A total of 22 differentially expressed miRNAs between A and B sandwiches were initially screened using gene chip technology,and miRNA-146 b,miRNA-29 b,and miRNA-21 with significantly different expressions were selected.Verified results by quantitative real-time PCR showed that miRNA-The relative expression levels of 146 b in group A,B,and C were 0.78±0.61,1.19±0.98,and 0.47±0.44,respectively.Compared with group C,the expression levels of group A and group B increased,and the differences were statistically significant.Significance(P<0.05 or P<0.01).Compared with group B,the expression of group B increased more significantly(P<0.05);miRNA-29 b was in A,B,and B.The relative expression levels in group C were 0.85±0.67,0.37±0.29,and 0.56±0.45,respectively.Compared with group C,the expression level in group A increased significantly,while the expression level in group B decreased significantly,and the differences were statistically significant.Significance(P<0.05 or P<0.01);relative expression levels of miRNA-21 in groups A,B,and C were 0.36±0.28,1.02±0.77,and 0.68±0.58,respectively,compared with group C,group A.The expression level was significantly reduced,while the expression level in group B was significantly increased,The differences were statistically significant(P<0.05 or P<0.01);both were in accordance with the initial screening results of the gene chip.The area under the ROC curve showed that the area under the curve of miRNA-146 b in the A and B groups was 0.653 and 0.607,respectively,and the area under the curve of miRNA-29 b in the A and B groups was 0.632 and 0.364,respectively.The area under the curve in the A and B groups was 0.301 and 0.661,respectively.According to the results,the sensitivity and specificity of miRNA-29 b and miRNA-21 in the diagnosis of the sandwich type were low,and the diagnostic criteria were not met;The area under the curve of miRNA-146 b is greater than 0.5,which has a certain diagnostic typing value.3.The bioinformatics analysis software was used to predict the downstream target genes of miRNA-146b: TRAF6,?B1,ACTA2,MMP16;the downstream target genes of miRNA-29 b were:TGF-?,Elastin(ELN),MMP2,MMP9,COL1A1,COL2A1,Downstream target genes for COL3A1,COL5A1;miRNA-21 are: PTEN.Conclusions1.There were significant differences in the expression of miRNA-146 b,miRNA-29 b,and miRNA-21 in plasma between acute Stanford type A and type B aortic dissections.2.Plasma miRNA-146 b has a certain diagnostic value for the classification of acute aortic dissection,which may be one of the potential biomarkers for its diagnosis.The differences in miRNA-29 b and miRNA-21 in acute Stanford A and B sandwiches The expression has not reached the diagnostic significance.Whether the two can be used as a marker for the diagnosis of AAD needs further study.3.The downstream target genes of miRNA-146 b,miRNA-29 b and miRNA-21 were preliminarily predicted and analyzed.It was speculated that the differential expression of these three in the acute Stanford A and B dissections was associated with the pathogenesis and prognosis of acute aortic dissection.