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血管内皮生长因子基因多态性与急性心肌梗死的相关性研究
摘 要
目的：急性心肌梗死(acute myocardial infarction，AMI) 冠状动脉急性、持续性缺血缺氧所引起的心肌坏死。临床上多有剧烈而持久的胸骨后疼痛，休息及硝酸酯类药物不能完全缓解，伴有血清心肌酶活性增高及进行性心电图变化，可并发心律失常、休克或心力衰竭，常可危及生命。血管内皮生长因子(vascular endothelial growth faetor，VEGF)是对血管内皮细胞有高度特异性的有丝分裂原，高浓度的VEGF可有效促进血管内皮细胞生长、分化及迁移，诱导新生血管
形成，促进体内组织建立起丰富毛细血管网；并且可增加毛细血管后静脉和小静脉通透性，而且还与心肌梗死患者免疫功能低相关。VEGF基因具有高度多态性，目前至少发现了30多个单核昔酸多态性(SNP)位点。本研究通过检测入选的急性心肌梗死患者的VEG血清浓度、VEGF的SNPs、生化、血常规、尿常规等，进行如下分析：1．VEGF基因SNPs与AMI发病风险的关联分析；2.血浆VEGF浓度与VEGF基因SNPs的关系；3.VEGF血浆水平与MACCE发生率相关性；4．LV- remodling与VEGF浓度及多态性的相关性；5.心肌梗死面积分别与VEGF血清浓度及VEGF的SNP的相关性。为急性心肌梗死的研究提供更多的临床数据参考。
方法：采用基于医院的病例一对照研究方法，将入选的患者分为急性心肌梗死患者组（研究组）和对照组，检测入选的急性心肌梗死患者的VEGF血清浓度、VEGF的SNPs、生化、血常规、尿常规；检测随访6月后的左室功能（左室重塑）；AMI发生6个月后再行如下检查：a．研究对象随访观测MACCE；b.行核素心肌显像测定心肌面积；c.行超声心动图测定左室重构。通过记录6个月内的心血管事件发生率。对比和研究以下数据：1．VEGF基因SNPs与AMI发病风险的关联分析；2.血浆VEGF浓度与VEGF基因SNPs的关系；3.VEGF血浆水平与MACCE发生率相关性；4．LV-remodling与VEGF浓度及多态性的相关性；5.心肌梗死面积分别与VEGF血清浓度及VEGF的SNP的相关性。
结果：通过对比和研究以下数据：1．VEGF基因SNPs与AMI发病风险的关联分析，结果表明VEGF基因SNPs与急性心肌梗死正相关（r=0.312）；2.血浆VEGF浓度与VEGF基因SNPs的关系成正相关(r=0.573)；3.VEGF血浆水平与MACCE发生率相关性分析表明，VEGF血浆水平、多态性与MACCE正相关（r=0.618）；4．LV- remodling与VEGF浓度及多态性正相关(r=0.187)；5.心肌梗死面积分别与VEGF血清浓度及VEGF的SNP的相关性结果表明，心肌梗死面积与VEGF血清浓度成正相关（r=0.736），心肌梗死面积与VEGF的SNP成正相关（r=0.582）。
结论：血管内皮生长因子的检测数据与急性心肌梗死其他检测的数据相关性分析表明，各数据间均呈正相关，对急性心肌梗死的血管内皮生长因子的检测对急性心肌梗死的研究诊断具有重要意义。
关键词：血管内皮生长因子；急性心肌梗死；单核苷酸多态性；血管生成；易感性
Abstract
Objective：Acute myocardial infarction (acute myocardial infarction, AMI) coronary acute myocardial ischemia and hypoxia caused by persistent necrosis. After more than a clinically severe and persistent chest pain, rest and nitrates can not complete remission, accompanied by increased serum enzyme activity and sexual ECG changes, may be complicated by arrhythmias, shock or heart failure, and often life-threatening. Vascular endothelial growth factor (vascular endothelial growth faetor, VEGF) is a vascular endothelial cells have highly specific mitogen, a high concentration of VEGF can effectively promote vascular endothelial cell growth, differentiation and migration, induce angiogenesis, and promote the body organizations Since the rich capillary network; and after the increase permeability of capillaries and venules veins, but also associated with lower immune function in patients with myocardial infarction. VEGF gene is highly polymorphic, currently found at least more than 30 single-core Xi acid polymorphism (SNP) sites. In this study, serum concentrations of selected VEG detection of acute myocardial infarction, VEGF of SNPs, biochemistry, blood, urine, etc., make the following analysis: 1. VEGF gene SNPs associated with AMI risk analysis; 2 between plasma concentration of VEGF and VEGF gene SNPs in; 3.VEGF plasma levels correlated with the incidence of MACCE; 4. LV- remodling polymorphism and VEGF concentration and correlation; 5 infarct size, respectively, serum concentrations of VEGF and VEGF SNP correlation. Provide more clinical data reference for the study of acute myocardial infarction.
Methods：A case-control study of hospital-based approach, the patient will be enrolled into patients with acute myocardial infarction (study group) and the control group, the serum concentrations of VEGF selected patients with acute myocardial infarction, VEGF of SNPs, biochemistry, blood, urine; detection followed up for 6 months after left ventricular function (left ventricular remodeling); AMI occurred six months after check the following: a. Study follow-up observations MACCE;. B-line determination of cardiac radionuclide myocardial perfusion imaging area;. C echocardiography left ventricular remodeling. By recording the incidence of cardiovascular events within six months. Compare and study the following data: 1. VEGF gene SNPs associated with AMI risk analysis; 2 between plasma concentration of VEGF and VEGF gene SNPs in; 3.VEGF plasma levels correlated with the incidence of MACCE; 4. LV-remodling polymorphism and VEGF concentration and correlation; 5 infarct size, respectively, serum concentrations of VEGF and VEGF SNP correlation.
Results：By comparing and research the following data: 1. VEGF gene SNPs associated with the risk of AMI analysis showed that VEGF gene SNPs are associated with acute myocardial infarction (r = 0.312);. 2 plasma concentration of VEGF and VEGF gene SNPs in relation to a positive correlation (r = 0.573); 3.VEGF plasma levels and the incidence of MACCE correlation analysis showed that, VEGF plasma levels of polymorphism and MACCE positive correlation (r = 0.618); 4. LV- remodling positive correlation with VEGF concentration and polymorphism (r = 0.187);. 5 infarct size were serum concentrations of VEGF and VEGF SNP correlation results showed that the myocardial infarct size and serum VEGF concentrations were positively correlated (r = 0.736), myocardial infarct size and VEGF SNP positive correlation (r = 0.582).
Conclusions：Vascular endothelial growth factor correlation analysis of the data and testing data to other detection of acute myocardial infarction showed a positive correlation between the data, the detection of vascular endothelial growth factor in patients with acute myocardial infarction has important significance for the study of the diagnosis of acute myocardial infarction.
Key words: Vascular endothelial growth factor; acute myocardial infarction; single nucleotide polymorphism; angiogenesis; susceptibility
目 录
引 言............................................................................................................................ 1 第一章 研究对象与方法.............................................................................................. 5
1.1研究对象.......................................................................................................... 5 1.2研究对象血标本收集及DNA提取 ............................................................... 6 1.3 仪器与试剂..................................................................................................... 8
1.3.1实验仪器............................................................................................... 8 1.3.2实验试剂............................................................................................... 9 1.4 外周血白细胞DNA的提取 ........................................................................ 10 1.5 DNA纯度的测定 .......................................................................................... 10 1.6 血标本的采集及DNA的提取 .................................................................... 19 1.7 统计学分析方法........................................................................................... 22 第二章 实验结果........................................................................................................ 24
1 两组研究对象的一般特征.............................................................................. 24 2 VEGF-460 T/C和- 1154G/A单核苷酸多态性分布与急性心肌梗死发病风险相关性分析.......................................................................................................... 24 3 PEDF基因各SNP位点等位基因频率的组间比较 ...................................... 27 4 eNOS基因SNP位点等位基因频率的组间比较 .......................................... 29 5 Ang-2基因SNP位点基因型及等位基因频率的组间比较 .......................... 30 6 相关SNP与ROP的logistic分析 ................................................................. 31 第三章 讨论................................................................................................................ 33
1 VEGF基因SNP与ROP发病的关联分析 .................................................... 33 2.本实验中釆用的研究方法和策略的探讨....................................................... 43 3 展望.................................................................................................................. 45 结论.............................................................................................................................. 56 参考文献...................................................................................................................... 57 综 述............................................................................................................................ 1 攻读学位期间的研究成果.......................................................................................... 19 致 谢.......................................................................................................................... 20