| Background: Coronary angiography technique, as the gold standard for diagnosis ofcoronary heart disease (CHD), belongs to invasive diagnosis with side-effect. Thenoninvasive64-slice CT and dual-source CT is widely used in the evaluation of patientswith lesions of coronary artery, but expensive. Clinical epidemiology survey showedthat the higher asymmetric dimethylarginine (ADMA) levels increased the dangerousfor cardiovascular disease. There is no method for ADMA detection meet therequirements of clinical testing, so ADMA is not applied in clinical. Our team is beingestablished ELISA kit for ADMA that can be used for clinical testing, and this studyestablished the determination method for ADMA with high performance liquidchromatography, in order to evaluate our ELISA assay kit and prepare for clinicaltesting. We hope to find out the different in35biochemical indicators in blood betweenCHD and Non-coronary heart disease (NCHD) and establish diagnostic model forscreening CHD, to reduce coronary angiography risk for NCHD.Material and Methods: According to the proposed included and excluded criteria, the398research objects were divided into CHD and non-CHD group according to coronaryangiography. The CHD group was divided into single-vessel CHD (SCHD) andmulti-vessel CHD (MCHD) group, which was divided into double-vessel CHD (DCHD)and three-vessel CHD (TCHD) group.The serum ADMA and symmetric dimethylarginine (SDMA) were detected withhigh-performance liquid chromatography,35conventional biochemical indicators withthe Roche COBAS8000automatic biochemical analyzer.For data analysis with SPSS19.0software, normal quantitative data was comparedby t test between the two groups and by one way ANOVA among multiple groups, and non-normality or heterogeneity of variance by non-parametric U test between two andby non-parametric H test among three. Qualitative data was analyzed with thechi-square test. P <0.05was considered statistically significant. The difference werescreened in35biochemical indicators for multivariate Logistic regression and stepwisediscriminant analysis, and mathematical classification model was established todistinguish between the CHD and NCHD group.Results: Arg, H-Arg, ADMA and SDMA in serum were detected with Highperformance liquid chromatography (HPLC), where the standard curve’s determinationcoefficient R2is more than0.999and its recovery range is from88.7%to110.2%andthe difference of two level recoveries was not statistically significant. The coefficient ofvariation with intra-assay and inter-assay was less than5%and7%, respectively. Thelimit of detection was0.02μmol/L for Arg, and0.01μmol/L for H-Arg, ADMA andSDMA, and the limit of quantification was0.05μmol/L for Arg, and0.03μmol/L forH-Arg, ADMA and SDMA. ADMA and SDMA in serum with CHD group wassignificantly higher comparing with NCHD group. The ADMA and SDMA had nostatistically differences among the CHD groups, but weakly associated with the numberof diseased coronary vessels and Gensini integral, respectively, and correlation test wasstatistically significant.Twelve routine biochemical indicators including ALT, GGT, TNT, UN, Cr, UA,APOA1, P, HDL, LP(a), Na and NTBNP in CHD group were significant statisticaldifferences, comparing with non-CAD group. TNT and NTBNP in MCHD group weresignificantly higher than in SCHD group. Binary logistic regression with SPSS19.0software showed that final five parameters into the equation yielded an AUC (areaunder curve) of0.766in discriminating group CHD from NCHD with sensitivity of83.4%and specificity of57.0%. Discriminant analysis showed that the same five parametersinto the discriminant equation yielded an AUC (area under curve) of0.762indiscriminating group CHD from NCHD with sensitivity of76.0%and specificity of66.7%. Conclusion: In conclusion, we established HPLC method for the determination of Arg,H-Arg, ADMA and SDMA in serum, and found that ADMA and SDMA concentrationin serum weakly correlated with the number of diseased coronary vessels and Gensiniintegral, respectively. The increasing ADMA or SDMA in blood may be risk factors forcoronary heart disease. In the study, the diagnosis equation with routine biochemicalparameters to distinct CHD from NCHD group was established, which could be used toscreen coronary heart disease. |
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