ABCA1基因启动子区域VNTR-ZNF位点多态性及与血浆HDL水平的关系

ABCA1基因启动子区域VNTR-ZNF位点多态性及与血浆

HDL水平的关系

高慎霞;毛用敏;陈倩;赵莉莉;崔让庄

【摘要】目的:探讨腺苷三磷酸结合盒转运体AI(ABCAI)基因启动子区域VNTR-ZNF位点多态性的分布频率及其与血浆高密度脂蛋白(HDL)水平的关系.方法:随机抽取166例冠心病患者和99例健康对照者,采用PCR扩增法和聚丙烯酰胺凝胶电泳对VNTR-ZNF位点ACCCC插入和缺失进行检测.结果:被检总人群ABCA1基因VNTR-ZNF位点等位基因频率分别为插人型为31.3%(166/530),缺失型

68.7%(364/530);基因型频率分别为插入型6.4%(17/265),缺失型

43.8%(116/265).插入/缺失型49.8%(132/265),2组间基因型频率和等位基因频率比较差异无统计学意义,血浆HDL及HDL亚组水平在3种基因型间差异亦无统计学意义(P>0.05).结论:天津地区ABCA1基因的VNTR-ZNF位点基因型分布与欧洲地区不同,此基因变异对血浆HDL水平无明显影响.%Objective: To investigate the frequency distribution of VNTR-ZNF polymorphism in ATP-binding cassette transponer 1(ABCA1) promoter region and the association with the plasma level of high-density lipoprotein (HDL). Methods:A total of 265 subjects were recruited, including166 patients with coronary heart disease (CHD) and 99 healthy control subjects. The corresponding DNA fragments were amplified with PCR and the genotypes were separated by polyacrylamide gel electrophoresis, detecting the insertion/deletion of ACCCC in VNTR-ZNF. Results: The allele frequencies of inserted form and deleted form of VNTR-ZNF were 31.3％ (166/530) and 68.7％ (364/530). The genotypic frequencies of VNTR-ZNF were 6.4％ (17/265) for the

inserted form, 43.8％ (116/265) for the deleted form and 49.8％ (132/265) for the inserted/deleted form.There were no significant differences in the frequency distribution of allele and genotype between CHD and control groups (P ＞ 0.05). Also there were no significant differences in levels of HDL and three different genotypes of HDL hetween two groups (P ＞ 0.05). Conclusion: The frequency of deleted form of VNTR-ZNF in ABCA1 in Tianjin district was markedly higher than that in European area, but the variation of ABCA1 VNTR-ZNF was not associated with the plasma level of HDL.

【期刊名称】《天津医药》

【年(卷),期】2011(039)005

【总页数】3页(P406-408)

【关键词】ATP结合匣式转运子;启动区(遗传学);脂蛋白类,HDL;冠心病;基因型;多态现象,遗传

【作者】高慎霞;毛用敏;陈倩;赵莉莉;崔让庄

【作者单位】300070,天津医科大学;天津市胸科医院;天津市胸科医院;天津市胸科医院;天津市胸科医院

【正文语种】中文

血浆高密度脂蛋白（HDL）水平降低是冠心病（CHD）发生的独立危险因素之一[1]。腺苷三磷酸（ATP）结合盒转运体A1（ATP-binding cassette trans⁃porter

A1，ABCA1）在启动胆固醇逆转运和HDL生成起始阶段发挥着关键作用[2]。ABCA1基因序列中某些核苷酸变异可改变其活性并影响HDL水平[3]。ABCA1基因启动子区域有多个转录因子的作用元件，其中-220 bp附近区域为ZNF202的

结合位点，此处有一个同向重复序列可变数（variable number of tandem repeats，VNTR）多态位点，为ACCCC插入缺失变异（VNTR-ZNF）。本研究

旨在对ABCA1基因启动子区域VNTR-ZNF位点插入/缺失多态性进行观察并探讨其与血浆HDL水平的关系。

1 对象与方法

1.1 研究对象随机抽取天津市胸科医院2007年9月—2009年4月住院的心肌梗死患者（按照WHO1979年诊断标准）166例（CHD组），其中男116例，女

50例，平均年龄（56.2±9.7）岁，均有持续性胸骨后剧烈疼痛，血清心肌酶升高及心电图ST-T变化。对照组99例，为同期本单位健康体检者，其中男71例，

女28例，平均年龄（51.9±7.4）岁。

1.2 方法（1）基因组DNA制备采用Triton低渗缓冲液溶血法从白细胞中提取。（2）PCR扩增引物参照文献设计[4]，上游引物：5′-CCC TAA GAC ACC TGC TGT ACCC-3；′下游引物：5′-CTG AGA ACC GGC TCT GTT GGT-3′。PCR（扩增体系反应体积为25 μL。各成分终浓度分别为：Tris-HCl 60 mmol/L，（NH4）2SO415 mmol/L，pH 9.5，MgCl2

2.5 mmol/L，脱氧核糖核苷三磷酸（dNTP）0.2 mmol/L，引物各0.4 μmol/L，Taq酶 2 .5 U，模板DNA 0.5 μg。循环温度：95℃5 min变性；95℃ 3 0 s、62℃ 3 0 s、72℃ 1 min，35个循环；最后72℃延伸7 min。（3）PCR扩增产物经8%聚丙烯酰胺凝胶电泳分离基因型，用凝胶

成像系统分析结果。（4）血脂测定：HDL及HDL亚组采用PEG20000进行沉淀去除低密度脂蛋白（LDL）和极低密度脂蛋白（VLDL），酶法测定上清液中高密

度脂蛋白胆固醇（HDL-C）水平。