冠心病心肌线粒体DNA5.0kb缺失的检测

目的探讨冠心病心肌线粒体DNA5.0kb缺失的检测。方法120例心脏的左、右心室肌各1份,分为正常对照组、病例相关组和病例组,每组40例80个样本。用断裂点连接PCR分别扩增样本含线粒体DNA5.0kb缺失的片段,巢式PCR检测含5.0kb缺失片段的扩增产物的准确性,半定量PCR对该产物进行定量,聚丙稀酰胺凝胶电泳检测PCR产物。结果在对照片段扩增成功的样本中,正常对照组未检出线粒体DNA5.0kb缺失;病例相关组检出2例,占6.07%(2/33);病例组检出29例,占85.29%(29/34);左、右心室肌线粒体DNA5.0kb缺失量分别为0.0015%~0.7813%和0.0008%~0.3906%。病例组与其他两组缺失率经χ2检验,其差异具有极显著性意义(P<0.001);左、右心室肌的缺失量经t检验,其差异具有极显著性意义(P<0.001)。结论冠心病心肌多有线粒体DNA5.0kb缺失,缺血明显的区域其缺失量也较高。

The detection of mitochondrial DNA 5.0Kb deletion of myocardium in coronary heart disease

Objective To discuss the detection of mitochondrial DNA 5.0Kb deletion of myocardium in coronary heart disease.Methods 120 myocardium cases were divided evenly into three groups: normal control, other related nonischemic diseases and coronary heart disease group. The myocardium of left and right ventricle were taken from each case for the study. GAP-PCR was performed to amplify the fragments containing mtDNA 5.0kb deletion, nested-PCR to detect the accuracy of GAP-PCR, semi-quantitative PCR to calculate the quantities ratio of the mtDNA 5.0kb deletion to the total mtDNA . The PCR products were detected by polyacrylamide gel electrophoresis.Results The ratios of the mtDNA 5.0kb deletion were recorded as the following: 0.00% in normal controls group, 6.07% (2/33) in other related nonischemic diseases group, 85.29% (29/34) in the coronary heart disease group. The ratios of mtDNA 5.0kb deletion of the left and right ventricular muscles were 0.0015%～0.7813% and 0.0008%～0.3906% respectively. Statistical analysis showed that there were significant differences in deletion ratios not only between the coronary heart disease group and other two groups (P<0.001), but also between left and right ventricle muscles (P<0.001). Conclusion Mitochondrial DNA 5.0kb deletion occurred in most myocardium of coronary heart disease. Furthermore, the severe the ischemic, the higher the deletion ratio.