The Diagnostic Value Of Serum ADAMTS1/4 In Patients With Type A AAD And Mechanism Research Of AngⅡ-Induced ADAMTS4 In Phenotype Transformation Of HASMC

Part oneThe diagnostic value of ADAMTS1/4 in patients with Type A AADObjective:Type A AAD,a serious cardiovascular emergency requiring urgent surgery,is the most common and serious AAD.Type A AAD possess a very high mortality rate with 1-2%per hour during the first 48 hours,which means about 50%die within 48 hours when left untreated,thus making early diagnosis and treatment critical for survival The aim of this study was to investigate the diagnostic value of AD AMTS1 and ADAMTS4 in patients with type A acute aortic dissection(AAD),and provide new method for the early diagnosis of type A AAD.Methods:Immunohistochemistry and qRT-PCR were used to evaluate the protein and mRNA expression levels of AD AMTS 1 and ADAMTS4 in 14 type A acute aortic dissection(AAD)tissues and 10 control aortic tissues.And the non-parametric rank correlation test was used to analyze the correlation between AD AMTS1 and ADAMTS4 protein levels in tissue and serum of patients with AAD.Serum AD AMTS1 and ADAMTS4 expression levels in 74 patients with type A AAD,36 patients with hypertension(HPT),and 34 healthy donors were examined by ELISA.The diagnostic value of serum ADAMTS1 and ADAMTS4 were determined by receiver operator characteristic curve(ROC).Furthermore,the dynamic change of serum ADAMTS1,ADAMTS4,D-dimer,and CRP were detected before and after surgery at different time-points in 14 patients with type A AAD.Results:AD AMTS1 and ADAMTS4 protein and mRNA expression levels were found to be significantly higher in 14 type A AAD tissues(p<0.0001)compared with 10 control tissues.Moreover,there was a positive correlation between AD AMTS 1 and ADAMTS4 protein in 14 A-type AAD aorta and serum.Serum AD AMTS1 and ADAMTS4 levels were significant higher in patients with type A AAD than those in the HPT and HD group(p<0.0001 for both).The AUC value,sensitivity,and specificity of ADAMTS1 were 0.9710(95%CI:0.9429 to 0.9991),87.84%,and 97.06%,respectively,and those of ADAMTS4 were 0.9893(95%CI:0.9765 to 1.002),94.59%,and 97.06%,respectively.In addition,serum ADAMTS4 level was gradually decreased with the time extension after surgery,similar with D-dimer change.Conclusion:Thus,these data suggest that measurement of serum AD AMTS 1 and ADAMTS4 levels could be potential diagnostic biomarkers for type A AAD,and ADAMTS4 might be a risk factor associated with type A AAD.Part twoMechanism research of AngⅡ-induced ADAMTS4 in PhenotypeTransformation、migration and proliferation of HASMCObjective:To explore how inflammatory factors can regulate the phenotype of aortic vascular smooth muscle cells and provide experimental evidence for subsequent related studies.Methods:1.The expression of ADAMTS4 mRNA and protein in HASMC was stimulated by different concentrations of cytokines(H2O2,TNF-α,TGF-β and Ang Ⅱ).2.Different concentrations of Ang Ⅱ were used to stimulate HASMC HASMC was used to detect the expression of OPN and SMA at mRNA and protein levels.3.The expression of OPN and SMA at mRNA and protein levels was detected by siRNA inhibition of ADAMTS4 expression.4.The expression of OPAM and SMA was detected by transwell assay.And the changes of p-Iκα,p65 and H3 in Ang Ⅱstimulated cells were detected by ADAMTS4 inhibition.Results:Compare with other inflammatory factors,Ang-Ⅱ can significantly increase the expression of ADAMTS4(P<0.05).Expression of ADAMTS4,OPN and SMA increased at the level of mRNA and protein with Ang Ⅱ stimulation(0-40nM)(P<0.05).Down-regulation of ADAMTS4 can significantly reduce the expression of OPN and SMA at the mRNA and protein levels in Ang Ⅱ-induced HASMC.Inhibiting ADAMTS expression can inhibit the migration and proliferation of HASMC and Ang Ⅱ-induced HASMC migration and proliferation.ADAMTS4 protein expression was inhibited,the addition of Ang Ⅱ results in the up-regulation of cytoplasm p65,and the nucleus p65 decreased.Conclusion:Ang Ⅱ can induce the phenotypic transformation of HASMC and the expression of ADAMTS4.The inhibition of ADAMTS4 expression can inhibit the expression of Ang Ⅱ-induced HASMC.The inhibition of ADAMTS4 can inhibit the migration and proliferation of Ang Ⅱ-induced HASMC.AngⅡ can induce IkBa phosphorylation Degradation,promote p65 from the cytoplasm to the nucleus transfer,induced ADAMTS4 expression.Part threeAnalysis of the clinical features of single center of acute aorticdissectionObjective:Aortic dissection(AAD)is a life-threatening disease that seriously compromises human health.This study was designed to investigate the epidemiological characteristics of AAD patients and provide a theoretical basis for early prevention of AAD.Methods:435 patients with AAD who had diagnosed and hospitalized for the first time in Wuhan University First Hospital(Hubei Provincial People’s Hospital)From January 2014 to December 2016 and their epidemiological and clinical data were gathered,and the epidemiological and clinical data of all patients were conducted retrospectively anaiysis.Results:① Among the 435 AAD patients,there were 345(79.31%)male patients and 90(20.69%)female patients,the ratio of male to female was nearly 4:1;②Among the 435 AAD patients,the high incidence of AAD was between 31 and 80 years old,accounting for 94.94%,and the peak incidence of AAD was between 51 and 60 years of age;③ The incidence of AAD was constant Seasonal,there were more patients in hospital in spring(January-March)and winter(10-12 months);④Among the 435 AAD patients,the number of Stanford Type A patients was 174,accounting for 40%,while Stanford B Type of patients were 261,accounting for 60%;⑤ Among the 435 patients with AAD,patients with a history of hypertension accounted for 64.59%(281 cases),Mafan syndrome patients accounted for 3.45%(15 cases),and only one patient had a history of hypertension and Mafan syndrome,patients with a history of coronary heart disease was 8.28%(36 patients).Conclusion:The probability of occurrence of AAD in males is higher than that in females.The peak incidence of AAD is between 51 and 60 years of age.Male patients with type A A and female patients with type A AAD;AAD occurs mainly in the spring and winter;...