| BackgroundCoronary heart disease (CHD) is seriously harmful to human's health. Though increasing new diagnostic methods are emerging, misdiagnosis is still difficult to avoid in clinical work. Exploring a simple and noninvasive way used to detect coronary heart disease and acute coronary syndrome (ACS) as early as possible is important for patients with CHD. Biomarkers in patients with CHD may have great value in the diagnosis of CHD because of their involvement or reflection of the occurrence or development of coronary atherosclerosis. Although a great deal of biomakers have been found in CHD patients, the value of them in the diagnosis of coronary atherosclerosis and ACS is not clear.ObjectiveBy measuring the plasma levels of high sensitivity C-reactive protein (hs-CRP), soluble CD40 ligand (sCD40L), monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8), and interleukin-6(IL-6), P-selectin, tissue-type plasminogen activator (tPA) in patients with different types of CHD, calculating the Gensini scores, and analyzing the relationship between the above biomarkers and CHD or ACS, we intended to clarify the value of them in detecting the existence and severity of coronary lesions.Materials and Methods1. SubjectsTotal of 170 subjects were enrolled in this study, including 129 cases of confirmed coronary artery disease by coronary angiography from February 2009 to January 2010 at Qilu Hospital of Shandong University, and 41 cases without coronary heart disease as the control group. There were 39 cases of SAP,43 cases of UAP and 47 cases of AMI.2. Diagnostic criteria1):Coronary artery disease:Patients were diagnosed to be coronary artery disease with at least one coronary artery of the left main, left anterior descending, left circumflex or right coronary artery showing stenosis more than 50% of its luminal diameter by coronary angiography; 2):SAP:Symptom of angina pectoris remains stable for more than a month.3):UAP:Patients who met any of the following criteria were diagnosed to be UAP:new onset of angina within one month, worsening of the previously present symptoms, with or without angina at rest, and with or without increased level of cardiac enzymes (less than twice of the normal upper limits for CK-MB); 4) AMI:Patients were diagnosed to be AMI who met any two of the following three criteria:persistent chest pain> 30min, dynamic ECG changes, myocardial enzyme CK-MB twice or more higher than the normal upper limits;3. Biomarkers measurementssCD40L, MCP-1, IL-8, IL-6, P-selectin, and tPA were measured by flow cytometric method in EDTA anticoagulated blood. Hs-CRP was measured by nephelometry in plasma content. The performance was strictly in accordance with the instructions.4. Gensini scores of coronary artery lesionsRefer to the reference (Gensini GG. A more meaningful scoring system for determining the severity of coronary heart disease[J]. Am J Cardiol,1983, 51(3):606-610.) [6]5. Statistical analysisData were analyzed on SPSS11.0. Continuous variables were expressed as mean±standard deviation (SD). Categorical variables were presented as ratio or constituent ratio. ANOVA was used to compare among multiple groups with continuous variables of normal distribution, in which LSD-t test was used for pairwise comparison; Rank-sum test was used for non-normal data, in which Mann-Whitney U test was used for pairwise comparison. Categorical variables were compared by chi-squared test; A two-tailed P<0.05 was considered to be statistically significant.Results1. Plasma levels of the tested biomarkers in each groupHs-CRP:Concentration of plasma Hs-CRP in SAP patients was no significant change compared with the control (p=0.085). It was significantly higher in UAP group and AMI group than in the control and SAP (P<0.00001, P<0.00001), but there was no difference between UAP group and AMI group.sCD40L:Plasma levels of sCD40L in SAP, UAP and AMI group all were significantly higher than that in control group (p=0.006, p<0.00001, p<0.00001); and plasma level of sCD40L in AMI group was higher than that in SAP group; There was no difference among the other groups.MCP-1:Plasma level of MCP-1 in each CHD group was higher than that in control group (p=0.048, p=0.026, p<0.00001 for SAP, U AP, AMI, respectively); and plasma levels of MCP-1 in AMI group was higher than that in UAP and SAP(p=0.027, p=0.009); but there was no difference in the latter two groups (p=0.802).IL-8:The plasma concentrations of IL-8 in all the CHD groups were significantly higher than that in the control group (p=0.004,P=0.002,P=0.001); There were no difference amongl the CHD groups.IL-6:The plasma concentrations of IL-6 in all the CHD groups were significantly higher than that in the control group (p<0.00001,p<0.00001,p<0.00001), The plasma concentration of IL-6 in AMI group was higher than that in SAP group (p=0.011);There were no difference among the other groups.P-selectin:The plasma concentrations of P-selectin in all the CHD groups were significantly higher than that in the control group (p=0.005,p=0.007,p=0.003), There were no differences among the CHD groups(p=NS). tPA:The plasma concentrations of tPA in all the CHD groups were significantly higher than that in the control group (p=0.001,p=0.003,p<0.00001); The differences in the three CHD groups were not significant (P=NS).2. Correlation between biomarkers and Gensini scoresThe linear correlation analysis between the biomarkers and the Gensini score showed MCP-1 (r=0.203, p=0.026) and IL-6 (r=0.322, p<0.00001) was positively correlated with the Gensini score. There were no relationship between other markers and Gensini scores.3. Correlations among biomarkersComprehensive relationship existed among the biomarkers in our study, especially the inflammatory marker IL-6, it correlated with all the other biomarkers(hs-CRP:r=0.373, p<0.00001;sCD40L:r=0.333, p<0.00001;MCP-1:r= 0.450, p<0.00001; IL-8:r=0.388, p<0.00001; tPA:r=0.373, p<0.00001; P-selectin:r=0.231, p=0.004)). There existed strong correlation between tPA and P-selectin(r=0.602, p<0.00001).4. Areas under ROC curve of each biomarker for diagnosis of CHDAreas under ROC curve of the six factors were obtained by taking the plasma values of each biomarker in the stable and unstable angina as the test variables, and coronary heart disease as the state variables. The result showed that the areas under ROC curve for sCD40L was 0.743 (p<0.00001), for MCP-1 was 0.674 (p= 0.008), for IL-8 was 0.641 (p=0.014), for IL-6 was 0.779 (p<0.00001), for tPA was 0.759 (p<0.00001), and for P-selectin was 0.675 (p= 0.003)。5. Logistic regression analysis of each variable for the prediction of coronary heart diseaseUnivariate Binary Logistic regression analysis of each variable for prediction of CHD showed that, for sCD40L,OR=1.490 (r=0.002); for MCP-1,OR=1.003 (p= 0.025);for IL-8,OR=1.339 (p= 0.025);for IL-6,OR=2.108 (p<0.00001); for tPA, OR=1.001 (p=0.001); for P-selectin, OR=1.017(p= 0.011. Multivariate logistic regression analysis showed only IL-6 had the role of diagnosing CHD,OR=1.857 (p=0.007)6. The value of the upper quartile of the tested biomarkers for the prediction of CHD By taking the upper quartile of each biomarker in the control group as the cut off value, we found that patients with IL-6 levels higher than the cut off value had six times risk of CHD (OR=6.490, p<0.00001);with sCD40L,6 times (OR=6.000, p<0.00001);with MCP-1, two times (OR=2.286, p=0.015);with IL-8, two times (OR=1.994, p=0.045);with tPA, six times (OR=5.802, p<0.00001); and with p-selectin,2 times (OR=1.841, p=0.017)7. The role of each biomarker in the prediction of ACSUnivariate logistic regression analysis for the risk of ACS showed OR=3.712 (p<0.00001) for hs-CRP; OR=1.654 (p<0.00001) for sCD40L; OR=1.004 (p=0.009) for MCP-1; OR=1.315(p=0.037) for IL-8; OR=2.017(p<0.00001) for IL-6; OR=1.001 (p<0.00001)for tPA; and OR=1.017 (p=0.007) for P-selectin. Multivariate logistic regression analysis only showed hs-CRP had the role in the prediction of ACS,OR=3.645 (1.909-6.962, P<0.00001).By taking the upper quartile of hs-CRP in the control group as the cut off value(cut off value in our study was 1.37), we obtained OR of hs-CRP for ACS was 11.273 (95%CI:4.726-26.870 p<0.00001), with a sentivity of 80.0%, specificity, 73.8%, positive predictive value,86.1%, and negative predictive value,64.6%.8. Follow-up20 cases of negative cardiovascular events occurred during follow up in our study, including 3 cases of death,1 case of recurrent myocardial infarction,2 cases of heart failure, and 14 cases of recurrent angina pectoris. Age and incomplete revascularization were the risk factor for cardiovascular events in our study. By logistic regression analysis, we got OR=1.147 (95%CI:1.000-1.476, p=0.050) for age, OR=1.498 (95%CI:1.115-2.253, p=0.005) for incomplete revascularization among gender, age, history of hypertension, history of diabetes, smoking, history of hyperlipidemia, the level of glucose, and patients'admission level of glucose, blood pressure, total cholesterol, low density lipoproteein cholesterol, high density lipoprotein cholesterol and the level of each biomarkers.Conclusions1. The plasma levels of sCD40L, MCP-1, IL-8,IL-6,tPA and P-selectin in CHD were higher compared with the control group,indicating they can reflect the existence of coronary atherosclerosis. Of the biomarkers, IL-6 had the greatest value for diagnosing CHD and had great clinical significance.2. The positive correlation of MCP-1 and IL-6 with Gensini scores showed in our study indicate the two biomarkers can reflect the severity of coronary artery disease.3. The positive correlation between IL-6 and other biomarkers suggests IL-6 is in the upstream of inflammatory cascade, and can influence the concentrations and functions of other biomarkers. By inhibiting the role of IL-6, we may reduce the inflammatory response of CHD and thus the progress of it.4. The higher positive correlation between tPA and P-selectin indicates the increase of tPA is a compensatory response.5. Hs-CRP is a very sensitive acute phase protein, and has a very important value for diagnosing ACS.6. Biomarkers in CHD patients undertaken revascularization as in our study don't have predicting role for major adverse cardiac events, while only age and incomplete revascularization are the risk factors for prognosis. |
PDF Full Text Request