本文關鍵詞：ABCB1基因多態(tài)性與氯吡格雷抵抗的相關性研究，由筆耕文化傳播整理發(fā)布。

        氯吡格雷和阿司匹林雙聯(lián)抗血小板治療，是急性冠脈綜合癥（Acutecoronary syndrome，ACS）和經(jīng)皮冠狀動脈介入術（Percutaneous coronaryintervention，PCI）后防治血栓性事件發(fā)生的基礎。但是不同個體對氯吡格雷的反應是多樣的，即使應用標準劑量的氯吡格雷，部分患者仍會發(fā)生心血管事件，稱為氯吡格雷抵抗（clopidogrel resistance，CR）。雖然現(xiàn)在尚不能完全闡明CR的機制，目前的研究發(fā)現(xiàn)CR的發(fā)生是由于多種因素共同導致的結(jié)果，而遺傳因素在CR的發(fā)生中可能發(fā)揮非常重要的作用。氯吡格雷在體內(nèi)需要經(jīng)過腸內(nèi)的吸收和肝臟的代謝才能發(fā)揮其抗血小板作用。ABCB1（ATP-binding cassette subfamily B member1）基因在氯吡格雷的腸內(nèi)代謝中發(fā)揮重要的作用。本研究擬采用聚合酶鏈反應（Polymerase Chain Reaction，PCR）及焦磷酸測序的分析方法，初步闡明ABCB1基因多態(tài)性與CR之間的相關關系。研究方法：本研究共入選500例住院冠心病患者，給予阿司匹林300mg以及負荷劑量氯吡格雷600mg或300mg，服藥后6小時（服用氯吡格雷600mg的病人）或24小時（服用氯吡格雷300mg的病人），用光學比濁法測定20μmol/LADP誘導的血小板聚集率，當20μmol/LADP誘導的血小板聚集率≥70%，定義為CR；<70%則定義為NCR。所有患者均采集外周血提取基因組DNA，從Hapmap數(shù)據(jù)庫選擇標簽單核苷酸多肽（tag single nucleotidepolymorphism，tagSNP），采用聚合酶鏈反應（PCR）和焦磷酸測序的方法，檢測所有入選研究對象ABCB1基因標簽SNP的單核苷酸多態(tài)性在CR組和NCR組的基因型和等位基因頻率分布。結(jié)果：(1) ABCB1基因共入選9個標簽SNP位點，分別為rs1922242、rs2235048、rs10808072、rs1989831、rs868755、rs4148733、rs1202184、rs1045642和rs13233308。(2)ABCB1基因SNP位點rs1045642（C3435T）在CR組和NCR組均存在CC、CT、TT三種基因型，在CR組和NCR組的基因型頻率分別為23.3%、46.0%、30.7%和37.7%、44.3%、18.0%。與C等位基因攜帶者相比，rs1045642SNP位點（C3435T）的T等位基因頻率在CR組明顯高于NCR組（53.7%vs40.1%，p<0.001）。(3) ABCB1基因其它SNP位點：rs1922242、rs2235048、rs10808072、rs1989831、rs868755、rs4148733、rs1202184和rs13233308等在CR組和NCR組間的分布頻率尚無顯著統(tǒng)計學差異（P>0.05）。結(jié)論：ABCB1SNP位點rs1045642（C3435T）可能是CR發(fā)生的一個獨立危險因素。

    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基因多態(tài)性與氯吡格雷抵抗的相關性研究

縮略語表5-7中文摘要7-9Abstract9-10前言11-14文獻回顧14-191. 材料與方法19-23    1.1 研究對象及材料19-232.實驗方法23-30    2.1 臨床資料的采集以及隨訪23    2.2 處理方法及測定指標23-25    2.3 連鎖不平衡特征和標簽 SNP 的選擇25-26    2.4 基因型檢測26-28    2.5 統(tǒng)計學分析28-303．實驗結(jié)果30-42    3.1 基因組 DNA 檢測及標簽 SNP 的測序結(jié)果30-34    3.2 研究對象的臨床特征34    3.3 Hardy–Weinberg 平衡檢驗34-35    3.4 ABCB1 基因標簽 SNP 的連鎖不平衡結(jié)構分析35-37    3.5 標簽 SNP 基因型以及等位基因頻率分布37-424．討論42-45小結(jié)45-46參考文獻46-53個人簡歷和研究成果53-54致謝54

  本文關鍵詞：ABCB1基因多態(tài)性與氯吡格雷抵抗的相關性研究，由筆耕文化傳播整理發(fā)布。