ABCB1基因多态性与氯吡格雷抵抗的相关性研究

  本文关键词：ABCB1基因多态性与氯吡格雷抵抗的相关性研究，由笔耕文化传播整理发布。

        氯吡格雷和阿司匹林双联抗血小板治疗，是急性冠脉综合症（Acutecoronary syndrome，ACS）和经皮冠状动脉介入术（Percutaneous coronaryintervention，PCI）后防治血栓性事件发生的基础。但是不同个体对氯吡格雷的反应是多样的，即使应用标准剂量的氯吡格雷，部分患者仍会发生心血管事件，称为氯吡格雷抵抗（clopidogrel resistance，CR）。虽然现在尚不能完全阐明CR的机制，目前的研究发现CR的发生是由于多种因素共同导致的结果，而遗传因素在CR的发生中可能发挥非常重要的作用。氯吡格雷在体内需要经过肠内的吸收和肝脏的代谢才能发挥其抗血小板作用。ABCB1（ATP-binding cassette subfamily B member1）基因在氯吡格雷的肠内代谢中发挥重要的作用。本研究拟采用聚合酶链反应（Polymerase Chain Reaction，PCR）及焦磷酸测序的分析方法，初步阐明ABCB1基因多态性与CR之间的相关关系。研究方法：本研究共入选500例住院冠心病患者，给予阿司匹林300mg以及负荷剂量氯吡格雷600mg或300mg，服药后6小时（服用氯吡格雷600mg的病人）或24小时（服用氯吡格雷300mg的病人），用光学比浊法测定20μmol/LADP诱导的血小板聚集率，当20μmol/LADP诱导的血小板聚集率≥70%，定义为CR；<70%则定义为NCR。所有患者均采集外周血提取基因组DNA，从Hapmap数据库选择标签单核苷酸多肽（tag single nucleotidepolymorphism，tagSNP），采用聚合酶链反应（PCR）和焦磷酸测序的方法，检测所有入选研究对象ABCB1基因标签SNP的单核苷酸多态性在CR组和NCR组的基因型和等位基因频率分布。结果：(1) ABCB1基因共入选9个标签SNP位点，分别为rs1922242、rs2235048、rs10808072、rs1989831、rs868755、rs4148733、rs1202184、rs1045642和rs13233308。(2)ABCB1基因SNP位点rs1045642（C3435T）在CR组和NCR组均存在CC、CT、TT三种基因型，在CR组和NCR组的基因型频率分别为23.3%、46.0%、30.7%和37.7%、44.3%、18.0%。与C等位基因携带者相比，rs1045642SNP位点（C3435T）的T等位基因频率在CR组明显高于NCR组（53.7%vs40.1%，p<0.001）。(3) ABCB1基因其它SNP位点：rs1922242、rs2235048、rs10808072、rs1989831、rs868755、rs4148733、rs1202184和rs13233308等在CR组和NCR组间的分布频率尚无显著统计学差异（P>0.05）。结论：ABCB1SNP位点rs1045642（C3435T）可能是CR发生的一个独立危险因素。

    Dual antiplatelet therapy (Asprin and Clopidogrel) is the cornerstone toprevent the occurrence of thrombotic events after acute coronary syndrome(ACS)and percutaneous coronary intervention(PCI). However, individuals may vary inthe responses to clopidogrel. Patients after receiving standard clopidogrel therapystill are at the risk of subsequent death and ischemic events,which definded asclopidogrel resistanse(CR). The mechanisms of CR have not been fullyelucidated and are most probably multifactorial, and genetic factors play animportant role in CR. Clopidogrel is a prodrug that requires absorption inintestinal and biotransformation in liver to an active metabolite. ATP-bindingcassette subfamily B member1(ABCB1) plays an important part in theabsorption of clopidogrel in vivo. In this study, we aimed to assess therelationship between ABCB1gene polymorphism and CR, by polymerase chainreaction(PCR) and sequencing analysis.Methods:500patients with coronary heart disease were involved who were treated with clopidogrel(600mg or300mg) and aspirin(300mg). Clopidogrelresponse was assessed by posttreatment ADP20μmol/L-induced plateletaggregation. Genomic DNA was extracted from peripheral blood. TagSNPs areselected form Hapmap. Clopidogrel resistance was defined by RPA (RPA≥70%).Genotypes were determined by polymerase chain reaction(PCR) and sequencinganalysis for tag single nucleotide polymorphism (tagSNPs) of ABCB1gene.Results:(1) There are9selected tagSNPs in the study, including rs1922242，，rs2235048，rs10808072，rs1989831，rs868755，rs4148733，rs1202184，rs1045642and rs13233308.(2) ABCB1rs1045642(C3435T）polymorphism were CC,CTand TT genotypes, in CR group and NCR group were23.3%、46.0%、30.7%and37.7%、44.3%、18.0%. T allele carriers in CR group are much more than those inNCR group（53.7%vs40.1%，p<0.001）.(3) ABCB1rs1922242，rs2235048，rs10808072，rs1989831，rs868755，rs4148733，rs1202184and rs13233308polymorphism were not significant correlated with CR (P>0.05).Conclusion: ABCB1rs1045642(C3435T）may be an independent risk factor forclopidogrel resistance.

        ABCB1基因多态性与氯吡格雷抵抗的相关性研究

缩略语表5-7中文摘要7-9Abstract9-10前言11-14文献回顾14-191. 材料与方法19-23    1.1 研究对象及材料19-232.实验方法23-30    2.1 临床资料的采集以及随访23    2.2 处理方法及测定指标23-25    2.3 连锁不平衡特征和标签 SNP 的选择25-26    2.4 基因型检测26-28    2.5 统计学分析28-303．实验结果30-42    3.1 基因组 DNA 检测及标签 SNP 的测序结果30-34    3.2 研究对象的临床特征34    3.3 Hardy–Weinberg 平衡检验34-35    3.4 ABCB1 基因标签 SNP 的连锁不平衡结构分析35-37    3.5 标签 SNP 基因型以及等位基因频率分布37-424．讨论42-45小结45-46参考文献46-53个人简历和研究成果53-54致谢54

  本文关键词：ABCB1基因多态性与氯吡格雷抵抗的相关性研究，由笔耕文化传播整理发布。