| Coronary arterial disease (CAD), and the most serious complications-myocardial infarction, become one of the leading causes of patient death, and how to reduce the incidence, disability and mortality of CAD has been the focus of research. As a critical illness, all of early detection, early diagnosis and early prevention are an ideal way to prevent the progressive development of CAD and to improve the cure and survival rate of the patients. Although the traditional methods, including ECG, coronary angiography and enzymatic indicators, are important to the CAD diagnosis, but they remain regrettable for the early detection of the disease. To do this, find and identify biomarkers for early diagnosis of CAD on the level of biochemistry and molecular biology become a hot topic of current research in the field.CAD is an inflammatory disease with multifactorial interactions such as immunization, environmental influence and genetic factors, and its incidence has obvious familial aggregation of a large number of genetic susceptibility genes, which play an important role in the development of diseases. The involving conditions are very complex, and the exact pathogenesis of this disease is still unclear, thus the effective diagnosis and treatment of CAD based on susceptible genes are limited to be discovered. miRNA, regulates widely gene transcription, was confirmed to act as a variety of roles in cardiovascular disease, or may reflect the susceptibility to cardiac and vascular injury, and the dependence for cardiovascular function in mammals. Currently, the main research focused on the relationship between miRNA and arrhythmia, myocardial hypertrophy, myocardial angiogenesis and apoptosis, and the mechanism of miRNA regulating the CAD system is also rarely reported. The TGF-β1is an important inflammatory cytokines in the human body, and shows significant role in the development of many inflammatory diseases, but there are different opinions on whether the content of serum TGF-β1is associated with susceptibility to CAD and related cardiovascular events. Therefore, this study firstly explored the relationship between TGF-β1/SMAD3pathways and CAD occurrence, further predict the upstream regulatory miRNA of TGF-β1/SMAD3, and reveals association of the target miRNA expression levels with susceptibility of CAD. The results will provide new biomarkers of clinical diagnosis and effective candidate prevention strategies for CAD.Object:1. Demonstrate the relationship between serum TGF-β1/SMAD3levels and CAD occurrence.2. Predict and identify TGF-β1/SMAD3target regulatory miRNA.3. Analyze association of circulation miRNA expression and CAD susceptibility.Methods:Firstly, serum TGF-β1and Smad3concentrations were evaluated in279patients with CAD and268controls without CAD or other interfering diseases. The clinical and biochemical characteristics of all subjects were also determined and analyzed by the statistical methods. Secondly, miRNA regulating targetedly TGF-β1/SMAD3were predicted by online bioinformatic tools from some databases including miRbase, Tarbase, targetScan, PicTar, PITA, RNA22, miRanda, MicroCosm, RNAhybrid and miRGen, and meanwhile other imRNA were selected through literature search. Taken together, the candidate miRNA were confirmed. At last, Subjects from2012to2013in Affiliated Hospital of Guangdong Medical College for treatment of patients with CAD group selected180cases,169cases of non-CAD group. The peripheral blood miR-1, miR-19a, miR-126, miR-133and miR-499expression levels were detected using real-time PCR, significant differences and logistic correlation analysis were carried out about grouping of disease type and stratification of risk factors, and ROC curve was used to compare with the current common clinical biochemical markers and evaluate the ability of every miRNA for CAD diagnostic value.Results:1. TGF-β1and Smad3concentrations in CAD patients were significantly higher than those in the controls. Meanwhile, serum TGF-β1content from AMI was significantly more than that in both SAP and UAP (P<0.05), while there was no marked contrast between SAP and UAP (P>0.05). However, Smad3levels showed no obvious difference among AMI, SAP and UAP. Therefore, TGF-β1and Smad3were potential biomarkers for CAD, and may be used to diagnose more accurately the presence of CAD than either Lpa, ApoA1, SUA, BUN or TG2. miR-1, miR-19a, miR-126, miR-133and miR-499were confirmed as the targets and prepared for the followed subsequent research.3. Circulation miR-1, miR-19a, miR-126, miR-133and miR-499expression in CAD group were higher than those in control, and diagnostic effect was superior to the current clinical biochemical index such as Lpa, BUN, Scr, SUA, Glu, CHOL, TG, HDL, LDL, ApoA1and ApoB.Conclusions:1. Serum TGF-β1and Smad3levels were closely associated with CAD occurrence, and even may become the useful biomarkers of diagnosis and stratification of CAD.2. miR-1, miR-19a, miR-126, miR-133and miR-499were the target miRNA of TGF-β1/SMAD3.3. There was close association of circulation miR-1, miR-19a, miR-126, miR-133and miR-499with CAD susceptibility, and can be used as the diagnostic targets.CAD is a serious cardiovascular disease, exploring the new susceptibility genes to create effective ways of CAD early diagnosis and prevention is an important strategy in the current study. In this paper, based on the large sample of CAD clinical resources, and a variety of techniques in biochemistry, bioinformatics and molecular biology, serum TGF-β1/SMAD3were confirmed as the factor closely associated with CAD occurrence, their targeted regulatoty miRNA (miR-1, miR-19a, miR-126, miR-133and miR-499) is more likely to be novel and effective diagnosis biomarker of CAD. |
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