| Objective:To explore the relationship between the expression of miR-192-5p in the serum and the presence of coronary heart disease(CHD),and to analyze the expression of miR-192-5p in ECs which were regulated by TNF-a and hypoxia.These studies provide evidence and research direction for further clarifying the pathogenesis of miR-192-5p in vascular endothelial cell injury of CHD.Methods:1.A total of 41 patients with coronary heart disease diagnosed by coronary angiography in the Department of Cardiology,General Hospital of Yanbian University from July 2017 to December 2017 were admitted as subjects of experimental group of study.We simultaneously recruited 40 health subjects without any evidence of CHD.All fasting subjects' venous blood were taken in the morning.MiR-192-5p expression was measured by real-time fluorescence quantitative PCR(qRT-PCR),and validate PCR products.We collected the basic characteristics and the clinical index of all subjects,and gather coronary angiography results of all patients.The relationship between the expression of miR-192-5p in the serum and the presence of CHD is analysised.2.Human umbilical vein endothelial cell(HUVEC)provided by ATCC is used in cell experiments.The 3-7 generation of HUVEC with a density of about 25%were inoculated into cell culture dishes,when the cell growth density reached 70-80%,they were transferred to ECM basal medium and continued to culture 12h.HUVEC were treated with 20ng/ml TNF-a and hypoxia to set time(12,24,36,48h).The cells were collected by PBS and the expression of miR-192-5p in endothelial cells was detected by real-time quantitative PCR.Cell experiments used endothelial cell damage model induced by TNF-a and hypoxia to simulate some cases of vascular endothelial injury in human body.3.Statistical analysis was performed with SPSS 17.0 statistics software.Categorical variables were expressed as percentages,and were analyzed by chi-square test.Measurement data were presented as x ± s,and metering data were compared using the variance analysis.Spearman correlation analysis was used in the correlation analysis between miR-192-5p and laboratory parameters.The risk of miR-192-5p contributed to the presence of CHD was estimated from logistic regression models,in which miR-192-5p expreesion level itself was used as a variable factor with or without adjustment for covariates.Receiver operating characteristic(ROC)curve analysis was used to assess the diagnostic accuracy.The area under the ROC curve(AUC)was used as diagnostic index.A p value of less than 0.05 was considered statistical significance.The reported P value were two-tailed in all calculations.Results:1.The results showed that the expreesion level of serum miR-192-5p was significant lower among CHD than that among healthy controls(P<0.001).2.The expression level of serum miR-192-5p in CHD significantly correlated with IL-6(r=-0.330,P=0.035).3.With adjustment for covariates,logistic regression analysis showed that the decreased level of miR-192-5p(adjusted OR=0.084,95%CI 0.013-0.534,P=0.009)was associated with risk of CHD.4.Receiver operating characteristic(ROC)curve analysis was used to assess the diagnostic accuracy.The AUC of miR-192-5p was 0.913(95%CI,0.846-0.979;p<0.001).The optimal cut-off value-1.73 of the relative level of miR-192-5p was a specificity of 97.5%and a sensitivity of 80.5%.5.In the TNF-? induced endothelial cell injury model,miR-192-5p Showed a downward trend,whereas at the time of 24 hour stimulated by TNF-a,the expression of miR-192-5p was significant lower than that from HUVEC exposed to vehicle alone(P=0.037),and there was no significant difference between other times(P>0.05).The results suggest that the expression of miR-192-5p was induced change in HUVEC by TNF-a.6.In hypoxia induced endothelial cell injury model,the expression of miR-192-5p in hypoxia treated was no significant difference between all times(P>0.05).The results suggest that it was not induced change in HUVEC by hypoxia.Conclusion:1.The expreesion level of serum miR-192-5p was significant lower among CHD than that among healthy controls.The decreased of miR-192-5p is associated with risk of CHD.Therefore,it is proposed to be considered as potential diagnostic biomarker for CHD.2.In the TNF-a induced endothelial cell injury model,the expression of miR-192-5p Showed a downward trend,it associated with times of exposed to TNF-a.The results suggest that the expression of miR-192-5p was induced change in HUVEC by TNF-a.3.In the hypoxia induced endothelial cell injury model,the expression of miR-192-5p was not significantly changed.4.The expression level of serum miR-192-5p in CHD was significantly negative correlated with IL-6,and miR-192-5p in endothelial cells was decreased by stimulating of TNF-a,while IL-6 was increased.The results suggest that it may involve in vascular endothelial inflammatory reaction and provide a new potential target in the treatment of coronary heart disease. |
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