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Objective: This study from peripheral blood of patients with acute myocardial infarction Exosomes quantitative and qualitative analysis to explore the correlation of patients with acute myocardial infarction Exosomes serum creatine kinase concentration of coronary artery disease extent. Learn more about the peripheral blood sources of Exosomes level in myocardial infarction role in the development of its forecast value of acute myocardial infarction. Methods: 1, clinical patient selection: 2010-9 to 2011-5 hospital Cardiology, 93 patients hospitalized for chest pain suspected coronary artery disease CAD selective coronary angiography. And 93 patients were divided into the following four groups: N group (24 cases, the normal control group), AMI group (23 patients with acute myocardial infarction), SA group (24 patients with stable angina pectoris), UA group (22 patients with unstable angina group). 2, peripheral blood the Exosomes the extract: in patients undergoing coronary angiography process to take blood 40ml, Ficoll plasma away from the experience, the same amount of PBS diluted 2000g * 30min centrifugation, the supernatant was transferred to transcend the tube 12,000 × g, centrifuged 45min, 4? C, then transferred to another one ultra from the tube 11,000 g * 2h, 4? C, 1ml PBS Resuspend complement PBS and filtered through a 0.22μm filter, 11,000 g * 70min after centrifugation Exosomes , 50μl-200μl PBS resuspended, and stored at -80 degrees refrigerator need to prepare for next experiments; 3, the BCA quantitative: Take the Exosomes samples 2UL, diluted 5-fold, in accordance with the sample and the substrate ratio of 1:20, in nanodrop BCA protein quantification; 4, flow cytometric analysis: 5ug of Exosomes with the 4μm diameter aldehyde / latex beads sulfate combined at room temperature for 15 min, add PBS added to 1ml room temperature shocks 2 h Add 1mol / L glycine, 30 min blocking reaction, 4000rpm centrifugal 3min, the supernatant was discarded, PBS/0.5% BSA and washed 3 times, 500 ul PBS/0.5% BSA resuspend and remove 10ul binding Exosomes magnetic beads were added to the PE-CD11c monoclonal antibody, PE-CD81 monoclonal antibody, the APC / cy7-MHC Ⅱ monoclonal antibody the, pacific bule-CD86 monoclonal antibody, PE-CD106 monoclonal antibody, PE-CD54 monoclonal antibody, FITC-CD3 monoclonal antibody, and the corresponding isotype control Incubate 1 h, PBS washed 3 times, machine detection [1]. 5, record the total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), glycated hemoglobin (HbA1c), CK, CK-MB, CK-MM, fasting plasma glucose (FPG), simultaneous measurement of systolic blood pressure (SBP), diastolic blood pressure (DBP), age, gender and other related indicators, ask a history of smoking, history of hypertension and history of diabetes. The severity of coronary lesion count and Gensini score evaluation of coronary lesions. 6, SPSS18.0 statistical software for analysis. Measurement data are expressed as mean ± standard deviation (x ± s). Compared using analysis of variance between groups of the two groups were compared using the t test; P lt; 0.05 for the difference was statistically significant. Two sample t inspection, diverse this mean was used to compare the variance analysis and comparison of the rate and the number of cases with the chi-square test (χ2 test); Exosomes with CK Gensini scores other single factor analysis applications Spearman rank correlation . Coronary heart disease risk factors were analyzed using Binary Logistic regression analysis, the regression model Gensini score as the dependent variable, age, total cholesterol, triglycerides, low-density lipoprotein, high density lipoprotein, white blood cell count, CD11C, CD86, MHC-DR , CK, CK-MM, CK-MB as the independent variable. P lt; 0.05 for the difference was statistically significant. Results: 1, AMI and UA patients Exosomes quantitative level significantly higher than the normal control group, the AMI group Exosomes the expression was significantly higher than the SA group, patients-Exosomes the phenotype analysis shows that AMI patients express CD11C, CD86, MHC-DR significantly higher than the normal group; 2, AMI and UA group Exosomes level, with Gensini scores were higher than the SA group, suggesting that its related to plaque stability of coronary lesions. Peripheral blood of Exosomes level and serum CK, CK-MB, CK-MM levels, WBC were positively correlated. , Peripheral Exosomes level increases with increasing Gensini score was positively correlated reflect Exosomes also associated with the severity of coronary atherosclerosis lesions. Conclusion: This study showed that the incidence and severity of peripheral blood of Exosomes with the acute myocardial infarction related. Peripheral blood of Exosomes with the serum CK, CK-MB level of CK-MM, Gensini integral, WBC were positively correlated. In addition to the enzyme levels and coronary angiography, peripheral blood Exosomes may be one of the indicators of the clinical assessment of myocardial infarction.
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