冠心病患者apoE基因调控序列-219(G/T)多态性及与血脂的关系

为了研究载脂蛋白E(apoE）基因调控序列－219（G／T）多态性与人类冠心病（CHD）及其血脂水平的关系，本文采用聚合酶链反应结合了限制性片段长度多态性方法。分析了108例健康人及86例冠心病患者的－219（G／T）基因型。PCR产物直接测序验下。我们发现冠心病组apoE－219（T／T）基因型频率（0．651）和T等位基因频率（0．767）分别显著高于对照组（0．444，0．653，P＜0．05）；冠心病组和对照组T／T基因型者血清总胆固醇高于G／G基因型者。这一结果提示apoE调控区基因多态性可能影响冠心病的发生。推测apoE－219（T／T）基因型是冠心病的危险因子之一。     

中国人内源性高甘油三酯血症载脂蛋白E基因多态性的研究

目的 探讨中国人内源性高城油三酯血症（endogenous hypertriglyceridemia,HTG)患者载脂蛋白E（apolipoprotein E,apoE)基因多态性及其与血脂和载脂蛋白水平的关系。方法 采用聚合酶链反应－限制性片段长度多态性分析方法,分别对225例HTG患者及230名血脂正常者的apoE基因型、空腹血脂及载脂蛋白AⅠ、AⅡ、B100、CⅡ、CⅢ、E进行了分析。结果 HTG患者的体重指数(BMI)` 清甘油三酯（TG）、总胆固醇（TC）、非高密度脂蛋白胆固醇（nHDLC)水平较对照组显著升高,血清高密度脂蛋白胆固醇（HDLC）则显著降低（P＜0．001）,并伴有载脂蛋白水平的异常。HTG组与对照组apoE基因型及等位基因频率分布均以E3／3和ε3最高,HTG组的ε2等位基因有增高的趋势（P＞0．05）。对照组ε2等位基因携带者血清TG和apoE水平较ε3和ε4等位基因携带者显著升高（P＜0．001）,其低密度脂蛋白胆固醇（LDLC）水平及apoE/ApoC Ⅲ比值则显著降低（P＜0．001）。结论 ε2等位基因与血清TG和apoE水平升高及LDLC水平降低有关,apoE/apoC Ⅲ比值降低可能与HTG患者血TG水平升高有关。     

载脂蛋白E基因多态性与新疆维吾尔族自然长寿的相关性研究

目的 探讨载脂蛋白E（apolipoproteinE，apoE）基因多态性与新疆维吾尔族自然长寿的关系。方法 应用聚合酶链反应-限制性片段长度多态性方法检测百岁组42名，90岁组102名，65～70岁组70名和对照组53名的apoE基因多态性。结果 百岁组apoE的ε3／3、ε2／3和ε3／4基因型频率分别为69．0％、23．8％和2．4％，其ε3、ε2和ε4等位基因频率分别为82．1％、16．7％和1．2％，百岁组ε3／4基因型及ε4、ε3等位基因频率显著低于对照组（P〈0．01），ε2／3基因型及ε2等位基因频率则显著高于对照组（P〈0．01）。百岁与opoE基因的ε2等位基因呈正关联，与ε4等位基因呈负关联。结论在新疆维吾尔族，opoE基因多态性与个体寿命密切相关，同时也应考虑到长寿是年龄依赖的多种因素影响的结果。     

α3—巨球蛋白基因多态性与Alzheimer病的关联研究

目的：观察α2-巨球蛋白基因（α2-macroglobulin,A2M) 内含子17一种五核苷酸缺失突变在广州地区汉族老年人中的分布,探讨其与晚发Alzheimer病（AD）的相关性。方法：以97例晚发AD患者和111名健康老年人为对照进行病例－对照研究。用聚合酶链反应－限制性片段长度多态性方法分析A2M缺失／插入多态性和载脂蛋白E（apolipoproteinE,apoE)基因多态性。结果：（1）A2M基因缺失突变在晚发AD患者中的频率为2．6％,在正常老年人中的频率为2．7％,在所有受试者中未检测到A2M突变纯合体,晚发AD患者和健康老年人之间不存在A2M等位基因和基因型分布的差异,A2M基因多态性与晚发AD无关联。（2）晚发AD患者中apoE等位基因ε4频率显著升高（Z＝3．32,P＜0．01）。晚发AD与ε3／ε4基因型正关联（RR＝2．62,χ^2=6.89,P＜0．01）,和等位基因ε4正关联（RR＝2．67,χ^2=10.71,P＜0．01）。（3）晚发AD无论是否伴有apoE-ε4均与A2M不存在相关性。结论：广州汉族人群中A2M基因缺失突变多态性与晚发AD不具有关联。     

中国汉族人芳香二烷基磷酸酯酶基因启动子多态性与冠心病的关系

芳香二烷基磷酸酯酶（paraoxonase,PON1)基因启动子区－108（C／T）多态性与人类冠心病（CHD）及其血脂水平的关系。采用多聚酶链反应－限制性长段多态性的分析方法（PCR－RFLP）检测CHD患者PON1基因启动子区－108位点的多态性。结果显示PON1启动子区－108位点存在多态性，出现三种基因型：TT，TC和CC。各等位基因的分布在正常对照组及CHD组之间存在显著性差异，且CC基因型的分布在两组间也有显著差异（P＜0．05）。正常对照组与CHD组间各基因型血浆Apo AI水平无显著性差异；CHD组CC纯合子的血浆高密度脂蛋白胆固醇（HDL－C）水平明显低于对照组（P＜0．05），而两组间TT纯合子和TC杂合子的HDL－C水平无统计学差异。说明该多态性可能与CHD有一定的相关性。     

阿尔茨海默病与载脂蛋白E基因-427C/T多态性的关联研究

目的 探讨上海地区汉族人群载脂蛋白E(apolipoprotein E，apoE)基因启动子区—427C／T多态性与Alzheimer病(Alzheimer's disease，AD)发病风险的关系。方法 采用聚合酶链反应和限制性片段长度多态性方法，在104例AD患者和110名正常人中检测了apoE基因—427C／T各基因型及基因频率的分布。按比值比(odds ratio，0R)作疾病关联分析。结果 (1)AD患者与正常对照人群之间不存在—427C／T各等位基因和基因型频率分布的差异(P＞0．05)；(2)按apoE ε4基因分层后，无论是ε4型人群还是非ε4人群都不存在AD患者与正常老人间多态分布的差异(P＞0．05)；(3)在—427C／T 3种基因型中，仅T／T型AD与apoE ε4等位基因呈正关联(OR＝3．94，95％CI：2．206—7．038，x^2＝21．48，P＜0．05)。结论 上海地区汉族人群中，apo E基因—427C／T多态不是AD的疾病易感因子。     

载脂蛋白E基因多态性与血脂代谢及冠心病的关系

目的：研究载脂蛋白E基因多态性对血脂代谢的影响及其与冠心病的关系。方法：运用PCR－PFLP方法检测168例江苏地区无血缘健康汉族人群。分析健康人群各基因型及等位基因对血脂、载脂蛋白及脂蛋白（a）的影响,同时测定63全冠心病患者载脂蛋白E基因型,并与性别相匹配的90例正常对照组比较各基因型及等位基因频率分布。结果：载脂蛋白E各基因型血清总胆固醇（TC）、低密度脂蛋白胆固醇（LDL－C）及载脂蛋白B（ApoB）水平由高到低依次为ε3／4＞ε3／3＞ε2／3;各等位基因TC、LDL－C及ApoB水平由高到低依次为ε4＞ε3＞ε2。ε2等位基因具有明显的降低TC、LDL－C和ApoB的作用,而ε4等位基因明显的升高TC、LDL－C和ApoB的作用。冠心病组ε4等位基因频率（12.70%）明显高于对照组（5.55%）。结论：载脂蛋白E基因多态性影响血脂及载旨蛋白水平。ε2 基因具有明显的降低TC、LDL－C和ApoB的作用,而ε4等位基因的作用正相反。ε4等位基因可能是冠心病的遗传易患因子。     

中国人内源性高甘油三酯血症与脂蛋白脂酶基因PvuⅡ多态性关联的研究

目的：探讨中国人内源性高甘油三酯血症（hypertriglyceridemics,HTG)与脂蛋白脂酶基因PvuⅡ多态性是否关联。方法：采用聚合酶链反应和限制性片段长度多态性方法,对成都地区135例内源性高甘油三酯血症患者和193名血脂正常者脂蛋白脂酶基因内含子6PvuⅡ多态性及其对血脂及载脂蛋白（apo)水平的影响进行了研究。结果：HTG患者和正常人均以P＋等位基因为主,HTG组以P＋P＋基因型为主,而正常对照组P＋P－基因型为主。HTG组的P＋P＋基因型分布频率及P＋等位基因分布频率则显著高于正常对照组（0.460vs0.337,P＜0.05;0.689vs0.565,P＜0.01)。P＋P＋基因型者的血清甘油三酯（TG）、apoCⅡ、apoCⅢ、apoE水平及TG／HDL－C比值较P－P－基因型者显著增高（P＜0．05）。结论：脂蛋白酯酶基因P＋P＋基因型与中国人内源性高甘油三酯血症的遗传易感性有一定关联。     

HMG-CoA 还原酶基因多态性与血浆血脂的关系

目的 探讨3—羟—3甲基戊二酰辅酶A(3—hydroxy—3—methylglutaryl coenzyme A，HMG—CoA)还原酶基因多态性在中国汉族人群中的分布及其与冠心病(coronary heart disease,CHD)的关系。方法 用聚合酶链反应—限制性片段长度多态性方法分析HMG—CoA还原酶基因第2内含子区ScrFl酶切多态性。结果 ScrFl多态位点等位基因A、a频率在CHD组和正常对照组分别为0．519、0．481和0．440、0．560。基因频率分布符合Hardy—Weinberg平衡定律。ScrF1酶切多态性基因型频率、等位基因A、a频率在组间比较差异无显著性(P＞0．05)，但是，基因型为AA的冠心病患者，其血浆极低密度脂蛋白、胆固醇水平显著高于其他基因型患者(P＜0．05)。中国人ScrFl多态位点A、a等位基因频率与欧洲白人比较差异有显著性(0．44vs0．55，0．56vs0．45，P＜0．05)，可能由于标本的种族来源不同所致。结论 ScrFl酶切多态性与CHD无相关性(P＞0．05)，但是，患者组AA基因型与血浆极低密度脂蛋白、胆固醇水平密切相关(P＜0．05)。     

2型糖尿病患者载脂蛋白e4等位基因与颈动脉中内膜厚度的关系

目的：了解2型糖尿病（diabetes mellitus,DM)患者载脂蛋白E（apolipoprotein E,apoE)基因型与颈动脉中内膜厚度的关系。方法：选择255例无血管并发症的2型DM患者和107名健康个体，采用聚合酶链反应等位基因特异寡核苷酸探针杂交技术检测其apoE基因型。结果：2型DM组与对照组apoE基因型及等位频率差异无显著性（P＞0．05）；两组中e4/4、e4/3亚组总胆固醇、低密度脂蛋白胆固醇、脂蛋白（a)水平明显高于e3/2、e2/2亚组（P＜0．05）；两组中e4/4、e4/3亚组颈动脉中内膜厚度明显高于e3/2,e2/2亚组（P＜0．05）。不论在对照组还是在2型DM ，当调整总胆固醇、低密度脂蛋白胆固醇、甘油三酯、脂蛋白（a)、血糖、年龄、体重指数、吸烟等因素的影响后，协方差分析结果显示，颈动脉中内膜厚度在这两个基因型亚组喑差异有显著性（P＜0．05）。结论：不论在健康人还是在2型DM患者，e4等位基因与早期颈动脉粥样硬化关联。     

Association of apolipoprotein E genotypes with serum lipid profiles in a healthy population of Taiwan

Apolipoprotein E (apoE, protein; APOE, gene) plays a major role in lipoprotein metabolism and lipid transport. Many investigators have described associations between apoE genotypes, coronary artery disease (CAD), and other risk factors. The aim of this study was to investigate the association between apoE genotypes and serum lipid profiles in a healthy population of 220 volunteers at Kaohsiung in Taiwan. Other CAD risk factors such as serum levels of apolipoprotein A-I (apoA-I), apolipoprotein B, (apoB), homocysteine (Hcy), folate, and vitamin B12 were also measured. ApoE genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In the study population, the frequency of apoE allele epsilon3 was greatest (85.2%); the frequency of epsilon2 was 8.4%; and that of epsilon4 was 6.4%. The serum apoA-1/apoB ratio showed significant difference among the 3 apoE genotype groups (p 0.0001); the apoA-1/apoB ratio was 1.9 +/- 0.1 (mean +/- SD) in the epsilon2 group, vs 1.4 +/- 0.04 and 1.5 +/- 0.12 in the epsilon3 and epsilon4 groups, respectively. No significant associations were found between APOE alleles and the serum levels of the various lipids or other CHD risk factors.     

Apolipoprotein E polymorphism in northwestern Greece: frequency and effect on lipid parameters

Apolipoprotein (apo) E gene polymorphism and its effect on serum lipid parameters were examined in a Greek population originating from northwestern Greece (n = 555). The allele frequencies were epsilon2: 6.3%, epsilon3: 80.7%, and epsilon4: 13.0%. The epsilon4 allele frequency was higher in our population than was previously reported in individuals from other parts of Greece. ApoE polymorphism was associated with significant differences in serum lipid, and lipoprotein levels. Particularly, individuals with the epsilon2 allele had higher serum triglyceride and apoE levels and lower levels of total cholesterol, low-density lipoprotein cholesterol, and apoB, compared to those with the alleles epsilon3 and epsilon4. However, the impact of the epsilon4 allele on lipid parameters seen in other populations was not observed in our population. Furthermore, the combination of apoE polymorphism and serum apoE concentration explained a larger percentage of serum lipid variability than the polymorphism alone. In conclusion, the results of our study suggest that ethnic differences, as well as alterations of serum apoE levels, significantly modify the relationship between apoE gene polymorphism and serum lipid variability.     

Apolipoprotein E gene promoter (–219G/T) polymorphism is associated with premature coronary heart disease

The relationship of two apolipoprotein (apo) E gene polymorphisms and coronary heart disease (CHD) was investigated in 118 Finnish families with premature CHD and in 110 healthy control subjects. Affected siblings and probands with premature CHD had higher frequencies of the T allele of the -219G/T promoter polymorphism and the epsilon 4 allele (genotypes epsilon 4/3 or epsilon 4/4) of the apo epsilon 2/epsilon 3/ epsilon 4 polymorphism than those of healthy control subjects. Additionally, when the two apo E gene polymorphisms were combined, affected siblings and probands had a higher frequency of the -219T allele and the epsilon 4 allele combinations than did healthy controls. The -219T and the epsilon 4 alleles both separately and together were associated with higher levels of 2-h glucose in an oral glucose tolerance test. These results indicate that the two polymorphisms of the apo E gene have similar effects on the risk of coronary atherosclerosis in families with premature CHD. This risk was not explained by the effect of apo E gene polymorphisms on cholesterol metabolism, but their effect on cardiovascular risk factor clustering with insulin resistance may be of importance. We conclude that in addition to the epsilon 4 allele, also the -219G/T promoter polymorphism of the apo E gene is associated with early onset CHD.     

Apolipoprotein E polymorphism and its relation to plasma lipids in coronary heart disease

Background : The present investigation is aimed at examining the Apolipoprotein E (APOE) genotypic influence on coronary heart disease (CHD) risk in northwest India (Punjab), where this disease is emerging as a major threat to public-health care system. Materials and Methods: The present study comprised of angiographically diagnosed coronary heart disease patients (n = 193) and controls (n = 150) of Punjab. Genetic polymorphism of APOE gene was investigated by polymerase chain reaction (PCR), and its association with lipid levels was evaluated. Results : The allele frequencies of epsilon2, epsilon3, and epsilon4 were 0.054, 0.795, 0.151; and 0.077, 0.856, 0.067 in patients and controls respectively. The bearers of E3/E4 genotype had threefold higher propensity of developing CHD in this population (OR, 3.04; CI, 1.55-6.25; P P P Conclusions : A significant association (P = 0.016) of epsilon4 allele, especially E3/E4 genotype, with CHD was observed, along with HDL-C and LDL-C concentrations, in the population of northwest India.     

高脂血症患者载脂蛋白E基因多态性与血脂水平分析

目的:分析载脂蛋白E基因多态性和高脂血症患者的血脂水平。方法:应用等位基因特异性多重PCR技术对高脂血症患者和健康对照者载脂蛋白E基因多态性进行分析,并测定所有样本血清载脂蛋白E等血脂指标水平。结果:高脂血症患者总胆固醇、甘油三脂、低密度脂蛋白胆固醇、载脂蛋白E水平明显高于健康对照组(P<0.05),而高密度脂蛋白胆固醇,载脂蛋白AI明显低于正常对照组(P<0.05);血浆中载脂蛋白E含量顺序是E2/3>E3/3>E3/4,两两比较具有统计学差异(P<0.05);在载脂蛋白E的基因型中以载脂蛋白E3/3型多见;高脂血症患者中载脂蛋白Eε4等位基因频率明显高于健康对照组(P<0.05)。结论:载脂蛋白Eε4等位基因与高脂血症有关,载脂蛋白E基因多态性可能是高脂血症患者的遗传因素。     

Qualitative and quantitative effects of APOE genetic variation on plasma C-reactive protein, LDL-cholesterol, and apoE protein

A genetic link between lipid metabolism and inflammation has been suggested by the association between variation in the APOE gene and plasma C-reactive protein (CRP). This association was confirmed among Caucasians and extended to an African-American population, and the well-known associations of APOE variation with LDL-C and apoE protein were also observed. While eight common variants in APOE were examined, the association with CRP involved primarily the two nonsynonymous SNPs that define the major epsilon2, epsilon3, and epsilon4 alleles. In particular, the strongest link involved lower CRP levels among carriers of the APOE epsilon4 allele that also contributes to the risk of cardiovascular and Alzheimer's diseases as well as to higher lipid levels. A lesser effect was characterized by lower CRP levels among carriers of a subtype of the epsilon3 allele. The magnitude of the association with plasma CRP was at least as great as the effect of variation in the CRP gene itself. Quantitative analysis suggested that the effect on CRP is more likely a consequence of intrinsic functional differences among the E2, E3, and E4 apoE proteins than different levels of apoE protein or LDL-C in the plasma.     

Apolipoproteins B and E, and angiotensin I-converting enzyme (ACE) genetic polymorphisms in Italian women with coronary artery disease (CAD) and their relationships with plasma lipid and apolipoprotein levels

The Xba I, Eco RI and the signal peptide insertion/deletion ( I/D ) polymorphic sites of APOB gene, the Cfo I polymorphic site of apolipoprotein E gene (APOE), and the insertion/deletion polymorphism of angiotensin I-converting enzyme (ACE) gene were studied using polymerase chain reaction (PCR) in 55 postmenopausal women with coronary artery disease (CAD) and in 119 control women of equivalent age. Patients and controls were recruited from the population of Rome, considered representative of Central and Southern Italy. There were no significant differences in allele frequencies between the two groups, though APOB X-, R- and I, APOE*3 , and ACE D alleles were slightly more frequent in the cases than in the controls. The patients did not differ from the controls for plasma total cholesterol (TC), HDL-cholesterol, LDL-cholesterol, and apoAI values, while they presented significantly higher levels of triglycerides and apoB, and lower apoE levels. TC, apoE, and apoB quantitative values, adjusted for age, varied significantly among APOB Xba I and APOE genotypes. APOB X-X-genotype was associated in patients with a significantly lower mean TC concentration than the other two genotypes pooled together. APOE 3–2 genotype in the controls had significantly lower TC levels with respect to the other two pooled genotypic classes and higher apoE levels compared to 3–3 and 4–3 genotypes. In the patients, 3–2 genotype had significantly lower apoB levels than the pooled 3–3 and 4–3 class. We conclude that in the Italian women the DNA polymorphisms studied in this work do not seem to be important risk factors for CAD occurrence; that apoE quantitation could be another useful parameter to identify subjects at risk of CAD; and that APOB X -and APOE*2 are the alleles that most influence the interindividual plasma lipid variation among CAD female patients.     

The importance of plasma apolipoprotein E concentration in addition to its common polymorphism on inter-individual variation in lipid levels: results from Apo Europe

Interindividual variation in the concentration of plasma lipids which are associated with coronary artery disease (CAD) risk is determined by a combination of genetic and environmental factors. This study investigates the effects of apoE genotype and plasma concentration on cholesterol and triglycerides (TG) levels in subjects from five countries: Finland, France, Northern Ireland, Portugal, and Spain. Age and sex significantly influenced serum cholesterol, TG and apoE concentrations. The age effect differs in males and females. The allele frequencies of the apoE gene, one of the most widely studied CAD susceptibility genes, were determined: the epsilon2 allele frequency and the apoE concentration showed a north-south increasing gradient while the epsilon4 allele frequency showed the reverse. ApoE plays an important role in lipid metabolism. Total cholesterol and TG concentrations were significantly dependent on apoE genotype in both sexes. These differences in lipids between genotypes were more pronounced when plasma apoE concentrations were taken into account.     

The use of measured genotype information in the analysis of quantitative phenotypes in man. II. The role of the apolipoprotein E polymorphism in determining levels, variability, and covariability of cholesterol, betalipoprotein, and triglycerides in a sample of unrelated individuals

Recent advances in molecular biology provide measures of genotypes at loci involved in lipid metabolism. Genotypes for apolipoprotein E (apo E) and quantitative levels of total plasma cholesterol, betalipoprotein, and triglycerides were measured in a sample of 223 unrelated individuals from Nancy, France. The frequencies of the epsilon 2, epsilon 3, and epsilon 4 alleles are 0.13, 0.74, and 0.13, respectively, in this sample. Significant differences among apo E genotypes were detected for these lipoprotein phenotypes. The average effect of the epsilon 2 allele was to reduce total plasma cholesterol and betalipoprotein levels by 0.52 mmol/L and 0.98, respectively, while the epsilon 4 allele raised these levels by 0.26 mmol/L and 0.61, respectively. Apo E genotype specific correlations suggest that this locus also has an effect on the coordinated metabolism between cholesterol and triglycerides. We infer that approximately 17% of the genetic variability in total plasma cholesterol may be attributable to this apo E polymorphism. No other single locus has been identified with such a large contribution to cardiovascular disease risk factors in the general population.     

高通量apoE基因分型技术的建立及应用

目的建立一种准确、快速、高通量的apoE基因分型技术。方法从外周血样品提取基因组DNA，PCR扩增覆盖第112和158密码子的apoE基因片段；构建apoE基因片段重组质粒，并进行定点诱变，以得到3种等位基因型的对照样品；PCR产物消化处理，以除去残余的引物和dNTPs；进行模板指导的荧光染料标记终止碱基的掺入反应，应用荧光偏振检测仪分析荧光偏振值的变化；检测79例阿尔茨海默病(Alzheimer’s disease，AD)患者和63名健康老年人的apoE基因型，分析基因型与AD易感性之间的关系。结果对分析结果进行测序验证，表明模板指导的荧光染料标记终止碱基掺入-荧光偏振检测技术分析结果与测序结果完全相符。AD组和健康对照组样品的基因分型结果提示apoEε4等位基因是迟发型AD的危险因素。结论应用此技术进行apoE基因多态性的基因分型分析，具有准确、简易和高通量等优势，可以作为AD风险分析的一种新技术．也适于apoE基因与其他疾病相关性研究时的大规模基因筛查分析。