Phosphodiesterase 4 D gene polymorphism in relation to intracranial and extracranial atherosclerosis in ischemic stroke

In ischemic stroke, extracranial MR angiography (ECMRA) is more frequently abnormal in Caucasians and intracranial (ICMRA) in Asians which may have a genetic basis. We report phosphodiesterase (PDE4D) gene polymorphism and its correlation with MRA findings in patients with ischemic stroke. Consecutive patients with MRI proven ischemic stroke undergoing MRA were included in this study. The severity of atherosclerotic stenosis on MRA was categorized into moderate 50%-80%, severe 80%-99%, and total occlusion 100% using NASCET criteria. The polymorphism in SNP 32, SNP 83 and SNP 87 of PDE4D gene was analyzed by PCR both in the patients and in 188 controls. Among the 148 patients, MRA was abnormal in 77% patients; ECMRA in 53.8%, ICMRA in 66% and both were abnormal in 42% patients. The frequency of CC genotype of PDE4D83 was significantly higher in the patients with ischemic stroke compared to controls (OR 3.38, 95% CI 1.61-7.11, P= 0.001). The frequency of TT genotype of PDE4D87 was significantly higher ICMRA abnormalities (20%) compared to normal ICMRA (2%). The genotype and allele frequency of PDE4D83 and PDE4D32 were not significantly related to MRA abnormalities. The role of PDE4D87 in atherosclerosis needs confirmation in larger studies.     

Associations of genotypes at the apolipoprotein AI-CIII-AIV, apolipoprotein B and lipoprotein lipase gene loci with coronary atherosclerosis and high density lipoprotein subclasses

Association studies were carried out in a sample of 86 patients from Sweden who had survived a myocardial infarction (MI) at a young age and 93 age-matched healthy individuals, to compare the impact of polymorphisms at the apolipoprotein (apo) AI-CIII-AIV gene cluster on among-individual differences in plasma lipid and lipoprotein traits, the five high density lipoprotein (HDL) subclasses (2b to 3c), lipoprotein lipase (LPL) activity and presence and progression of atherosclerosis. Individuals were genotyped for four polymorphisms; 5'apoAI (G/A_{_75}), 3'apoAI (PstI: P ±), apoCIII (C/T₁₁₀₀) and apoCIII (PvuII; V ±), using PCR-based techniques. Allele frequencies were similar in healthy individuals and patients (frequencies of alleles in combined population: 5'apoAI-A_-75=0.14, 3'apoAI-P-=0.05, apoCIII-T₁₁₀₀=0.27 and apoCIII-V-=0.18). In the healthy individuals, levels of low density lipoprotein (LDL) triglycerides were significantly associated with genotypes of the apoCIII-PvuII polymorphism (p=0.02), but no other associations were found between lipids or HDL subclasses and single polymorphisms in the apoAI-CIII-AIV gene cluster. Levels of triglycerides and very low density lipoprotein (VLDL) triglycerides were significantly higher in the presence of the haplotype defined by the presence of apoCIII-T₁₁₀₀ and common alleles of the other three polymorphisms, explaining 5.8% and 7.8% (p=0.03 and 0.01), respectively, of sample variance. In the patients, no associations were found between lipids or HDL subclasses and variation at the apoAI-CIII-AIV gene cluster. Associations were also examined between levels of HDL subclasses and variation at the apoE (common isoforms), apoB (signal peptide and XbaI polymorphisms) and lipoprotein lipase (PvuII, HindIII and Serine₄₄₇/Stop polymorphisms) gene loci. In the patient group only, levels of protein in HDL2b, HDL2a and HDL3b subclasses were significantly associated with genotypes of the LPL-HindIII polymorphism (22.1, 19.3 and 11.4%, respectively, of sample variance; p < 0.05). Finally, associations were examined between genotypes at the apoAI-CIII-AIV gene cluster and the extent of coronary atherosclerosis. Global severity of atherosclerosis at the first angiography was weakly associated with genotypes of the apoCIII-C/T₁₁₀₀ polymorphism, presence of the T₁₁₀₀ allele being associated with 53% lower median score (1.6 vs 0.75; p = 0.09). In this group of patients, two genotypes, one each at the LPL and apoB gene loci, had been previously found to be associated with high atherosclerosis score and, when considered together, individuals with all three of these genotypes had the highest median score (2.4) and those with none of these genotypes had the lowest (0.4) (chi-squared overall = 15.7; p = 0.001); no lipid traits measured showed a similar association with these genotypes. Thus, in this sample of young male post-infarction patients, genetic variation at these three loci is having an additive effect on the development of atherosclerosis, that cannot be explained by their observed effect on fasting lipid and lipoprotein traits.     

The key role of apolipoprotein E in atherosclerosis

Apolipoprotein E is a multifunctional protein that is synthesized by the liver and several peripheral tissues and cell types, including macrophages. The protein is involved in the efficient hepatic uptake of lipoprotein particles, stimulation of cholesterol efflux from macrophage foam cells in the atherosclerotic lesion, and the regulation of immune and inflammatory responses. Apolipoprotein E deficiency in mice leads to the development of atherosclerosis and re-expression of the protein reduces the extent of the disease. This review presents evidence for the potent anti-atherogenic action of apolipoprotein E and describes our current understanding of its multiple functions and regulation by factors implicated in the pathogenesis of cardiovascular disease     

载脂蛋白E基因多态性与术后谵妄的关系

目的 探讨载脂蛋白E(APOE)基因多态性与老年非心脏手术患者术后谵妄是否具有相关性。方法 212例65岁以上的择期非心脏手术患者纳入研究,于手术后1~3d密切随访,根据CAM的标准判断患者有无发生谵妄。用突变特异性多重扩增系统 (multi-ARMS PCR)方法测定患者APOE基因型。结果 212例患者中有45例发生术后谵妄,共检出APOEε4等位基因携带者18例(8.5%)。谵妄组有3例APOEε4携带者（6.7%）,非谵妄组有15例APOEε4携带者（9.0%）,两组比较无显著差异。ε4/4纯合子型共有4例,其中1例术前3d发生过一过性谵妄,还有1例发生术后严重谵妄,症状持续17d。结论APOEε4等位基因与术后谵妄发病率无显著的相关性,但ε4/4纯合子型可能更容易发生谵妄。     

Genetics of atherosclerosis: Some strategies for studies of apolipoprotein E

Many genes are hypothesized to be involved in determining an individual's risk for coronary artery disease (CAD). Recent efforts have focused on the genetic architecture of quantitative traits related to risk for CAD. Studies relating genetic variation in the structural gene for apolipoprotein E to plasma levels of apolipoprotein E illustrate strategies to begin to understand the genetic architecture of plasma levels of apolipoprotein E. Studies using a measured gene approach suggest that variability in the isoforms of apolipoprotein E explain some, but not all, of the variability in plasma levels of apolipoprotein E. Products of other genes may be associated with plasma apolipoprotein E variability. No studies to date have presented findings from an unmeasured gene approach to plasma levels of apolipoprotein E. Models most often used in the unmeasured gene approach are not appropriate for studies of plasma levels of apolipoprotein E and perhaps are inappropriate for the study of other traits. Variations of the models are suggested to ask if a single unmeasured gene is the same gene defined by the apolipoprotein E isoforms. Another model can be used to ask if there is evidence for genes influencing levels of apolipoprotein E after accounting for the effects of the isoforms. Both the measured gene and unmeasured gene strategies have limitations. The failure of most models to allow for the complexity of genotype-phenotype relationships or the heterogeneity of genetic causes will slow the progress to understand the genetic architecture of quantitative traits associated with risk for CAD. © 1993 Wiley-Liss, Inc.     

载脂蛋白E基因多态性与疾病的相关性研究

载脂蛋白E（ApoE）是一种重要的血浆脂蛋白,由3种等位基因构成：E2、E3和E4。ApoE作为一种载脂蛋白,在脂质运输和代谢过程中发挥重要作用,而目前越来越多的研究表明：ApoE在免疫调节方面发挥重要作用,从而参与到多种疾病的发生发展中。近年来发现,ApoE及其基因多态性与高脂血症、动脉粥样硬化、Alzheimer病、神经系统病变及脓毒血症等人类疾患的发生发展有着密切关系。     

Association of apolipoprotein E gene polymorphism with essential hypertension and its complications

The clinical significance of the apolipoprotein E genotype in patients with hypertension has been a subject of debate. We enrolled 94 patients with hypertension and 102 healthy controls in this study and determined their plasma levels of triglyceride, total cholesterol, high- and low-density lipoprotein-cholesterol, apolipoprotein AI, and apolipoprotein B. The apolipoprotein E genotypes were identified by polymerase chain reaction, restriction fragment length polymorphism, and polyacrylamide gel electrophoresis. Apolipoprotein E3/4 genotype and 4 allele frequencies in the hypertensive group were higher than in controls. In hypertensive patients with apolipoprotein E3/4 and E4/4 genotypes, systolic blood pressure was significantly higher than in those with apolipoprotein E2/3 or E3/3 genotypes. Meanwhile, the plasma levels of total cholesterol, low-density lipoprotein-cholesterol, and apolipoprotein B were higher in hypertensive patients with the .4 allele than the 2 or 3 allele. The echographic measurements of carotid artery intimal-medial thickness showed increasing values from 2 to 4 allele carriers in the hypertensive group. Analysis of variance showed that the carotid intimal-medial thickness was significantly greater in hypertensive patients with 4 alleles compared with 2 or 3 alleles. Our data show an association between apolipoprotein E genotype and hypertension and support the hypothesis that the apolipoprotein 4 allele is a susceptibility locus for systolic hypertension and carotid artery atherosclerosis. Received: 23 July 2002 / Accepted: 27 December 2002 Correspondence to Xiaotao Li     

Matrix metalloproteinase-3 gene polymorphisms are associated with ischemic stroke

Stroke is a heterogeneous disease caused by different pathogenic mechanisms. Several candidate genes for stroke have been proposed, but few have been replicated. Matrix metalloproteinases (MMPs) are expressed following stroke. We investigated the association of single nucleotide polymorphisms (SNPs) of the MMP3 gene with stroke in the Korean population. This study included 186 stroke patients [116 ischemic stroke (IS) and 70 intracerebral hemorrhage (ICH)] and 668 age-matched control subjects (267 for IS and 401 for ICH). Three SNPs [rs520540 (Ala362Ala), rs602128 (Asp96Asp), and rs679620 (Lys45Glu)] in the coding region of MMP3 were selected and genotyped by direct sequencing. HelixTree, SNPAnalyzer, SNPStats, and Haploview version 4.2 were used to analyze genetic data. Multiple logistic regression models (codominant, dominant, and recessive models) were conducted to evaluate odds ratio, 95% confidence interval, and P value. Three SNPs in the MMP3 gene were significantly associated with IS (P<0.05). The genotype distribution of 3 SNPs differed between the IS and control subjects. However, there was no association of the SNPs between the ICH and control. In analysis of gender, 3 SNPs were also associated with IS in female group (P<0.05). These SNPs remained significantly associated with IS after the Bonferroni correction for multiple testing (P(c)<0.05). Haplotype analysis revealed that no haplotypes were associated with IS or ICH. Overall, the results of our study demonstrate an association of the MMP3 gene with development of IS, and no association of MMP3 with ICH.     

非洛地平对高胆固醇饮食载脂蛋白E基因敲除小鼠动脉粥样硬化的影响

目的：探讨钙通道阻断剂（CCB）非洛地平对载脂蛋白E基因敲除(ApoE KO)小鼠动脉粥样硬化斑块的影响。方法:ApoE KO小鼠随机分为普食组、高胆固醇饮食组、高胆固醇饮食＋非洛地平组（n=15），分别予蒸馏水或非洛地平 5 mg·kg-1·d-1灌胃12周。无创血压系统测小鼠血压；内眦动脉取血检测血清胆固醇和甘油三酯水平；冰冻切片光镜下定位主动脉根部，油红O染色评估斑块大小；实时定量PCR和Western blotting方法检测主动脉中炎症因子表达。结果:高胆固醇饮食组小鼠血脂明显升高（P<0.01），且斑块面积明显高于普食组（P<0.01）；非洛地平可以明显减小斑块面积（P<0.01），同时还可以降低肿瘤坏死因子-α（TNF-α）、单核细胞趋化因子-1 （MCP-1）和血管细胞黏附因子-1（VCAM-1）的表达。结论:钙通道阻断剂非洛地平可能通过抑制炎症反应，降低炎症因子表达，从而达到抑制动脉粥样硬化发生发展的目的。     

载脂蛋白E基因多态性与散发性老年性痴呆病的关系

目的:探讨载脂蛋白E(apoE)外显子4和增强子元件基因多态性与散发性Alzheimer病(AD)的关系。方法:应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术分别检测apoE外显子4和内含子1内增强子元件(IE1)基因型。结果:(1)ApoE外显子4基因多态性:AD组ε3/4基因型频率(0.381)和ε4等位基因频率(0.226)显著高于对照组(P＜0.05)。(2)ApoEIE1基因多态性AD组G/G基因型频率(0.595)显著高于对照组(P＜0.05)。(3)有apoEε4个体患AD风险为无ε4个体的3倍,比值比为2.932,95%可信区间1.379~6.226;G/G基因型个体患AD风险为G/C、C/C个体的2倍,比值比为2.223,95%可信区间1.075~4.599;经统计分析发现apoEε4与IE1G/G呈非常显著性正相关(P＜0.01);排除apoEε4后发现IE1G/G与AD发病风险无关。结论:ApoEε4等位基因是个体发生AD的危险因素,IE1G/G增加AD发病风险是因其与ε4相关所致。     

整合素基因多态性与缺血性脑卒中及血脂的相关研究

目的 探讨整合素-α2基因(integrin alpha-2,ITGA2)C807T和整合索-β3基因(integrinbeta-3,ITGB3)T176C多态性与缺血性脑卒中的关系及其对血脂、脂蛋白水平的影响.方法 应用聚合酶链反应-限制性片段长度多态性和DNA测序的方法检测265例缺血性脑卒中患者和280名对照组ITGA2和ITGB3的基因型;同时按常规方法测定血浆脂质、脂蛋白水平.结果 缺血性脑卒中组总胆固醇(totalcholesterol,TC),甘油三酯(triacylglycerol,TG)、低密度脂蛋白-胆固醇(low density lipoprotein-cholesterol,LDL-C)水平明显高于对照组(P<0.05),ITGB3基因T176C多态性在缺血性脑卒中组和正常人群中的分布差异无统计学意义(P>0.05).而ITGA2基因C807T多态性在两组人群中的分布差异有统计学意义(P<0.05),等位基因频率的相对风险分析发现,T等位基因携带者患缺血性脑卒中的风险是C等位基因的1.455倍(OR=1.455,95%CI:1.134～1.866),携带T等位基因的缺血性脑卒中个体血浆TC水平显著高于不携带者(P<0.05).结论 ITGA2基因C807T多态性与缺血性脑卒中的发病具有相关性,其中T等位基因可能是缺血性脑卒中的遗传易感基因;ITGA2基因C807T多态性可能通过影响血脂水平而影响缺血性脑卒中的发生.     

The apolipoprotein E gene in Binswanger's disease and vascular dementia

We investigated the polymorphism of the apolipoprotein E (ApoE) gene using a PCR-RFLP method in patients with Binswanger's disease (BD), non-BD vascular dementia, or Alzheimer's disease (AD). The frequency of the e4 allele of the ApoE (ApoE4) in BD patients and non-BD vascular dementia patients did not differ from that observed in the non-demented elderly controls, but it was significantly lower than the frequency in AD patients. These results and other recent observations suggest that one or more factors other than the ApoE gene contribute to the pathogenesis of dementia in BD and non-BD vascular dementia.     

构建不同周龄载脂蛋白E基因敲除动脉粥样硬化模型小鼠血脂和病理学观察

背景：载脂蛋白E基因敲除小鼠形成的动脉粥样硬化病变与人类全身动脉粥样硬化好发处相近,是目前建立动脉粥样硬化理想的动物模型。 目的：研究载脂蛋白E基因敲除小鼠不同周龄动脉粥样硬化的病理进程,探讨不同饮食对载脂蛋白E基因敲除小鼠动脉粥样硬化发生发展的影响。 方法：将8周龄雄性载脂蛋白E基因敲除小鼠,随机分为2组,分别给予高脂饮食和普通饮食喂养8,12,16,20,24周。 结果与结论：血清学指标检测显示,不同周龄的高脂饮食组血清中总胆固醇、三酰甘油和低密度脂蛋白胆固醇水平显著高于普通饮食组(P < 0.05),呈时间依赖性。大体和冰冻切片油红O染色结果显示,高脂饮食组动脉粥样硬化管腔斑块面积显著高于普通饮食组(P < 0.05),呈时间依赖性,此时两组各周龄小鼠管腔斑块面积相比均有显著性意义(P < 0.05),小鼠在高脂饮食16周时主动脉可见明显的脂质斑块。结果表明,实验成功构建了载脂蛋白E基因敲除动脉粥样硬化模型小鼠,此模型形成脂质条纹和纤维增生病变的时间较普通饮食组更快。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

The apolipoprotein E gene, attention, and brain function

The epsilon4 allele of the apolipoprotein E (ApoE) gene is associated with alterations in brain function and is a risk factor for Alzheimer's disease (AD). Changes in components of visuospatial attention with ApoE-epsilon4, aging, and AD are described. Healthy middle-aged adults without dementia who have the ApoE-epsilon4 gene show deficits in spatial attention and working memory that are qualitatively similar to those seen in clinically diagnosed AD patients. The findings support an association between ApoE polymorphism and specific components of visuospatial attention. Molecular mechanisms that may mediate the ApoE-attention link by modulating cholinergic neurotransmission to the posterior parietal cortex are discussed. Studies of attention and brain function in ApoE-epsilon4 carriers without dementia can advance knowledge of the genetics of visual attention, may enhance understanding of the preclinical phase of AD, and may lead to better methods for early AD detection.     

基质金属蛋白酶-3基因多态性与脑卒中亚型的相关性

目的 探讨中国北方汉族人群基质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)基因多态性与缺血性脑卒中(ischemic stroke,IS)亚型的相关性.方法 应用病例对照研究,选取289例急性缺血性脑卒中患者(发病≤3d)和175名同期健康体检者.卒中组按急性卒中治疗低分子肝素试验病因分型法分为大动脉粥样硬化性(large artery atherosclerotic,LAA)脑卒中185例,小动脉闭塞性(small artery occlusion,SAO)脑卒中104例.选取MMP-3基因rs3025058(-11715A/6A),rs522616(-709A/G)及rs679620(133A/G)3个常见单核苷酸多态性(single nucleotide polymorphisms,SNPs)位点,应用聚合酶链反应限制性片段长度多态性或直接测序法对MMP-3基因3个SNP位点进行基因分型.结果 3个位点存在不完全连锁不平衡,且基因多态性均与LAA型脑卒中相关(P＜0.05).在显性模型中,rs3025058位点5A5A+5A6A基因型携带者患LAA型脑卒中的风险是6A6A基因型携带者的1.72倍(P=0.017,OR=1.72,95％CI:1.10～2.69);rs522616位点GG+ AG基因型携带者患LAA型脑卒中的风险是AA基因型携带者的0.52倍(P=0.005,OR=0.52,95％CI:0.33～0.82);rs679620位点AA+ GA基因型携带者患LAA型脑卒中的风险是AA基因型携带者的1.55倍(P=0.042,OR=1.55,95％CI:1.01～2.37).但是,3个SNPs基因型和等位基因频率在对照组与SAO型脑卒中之间差异无统计学意义(P＞0.05).另外,LAA组5A-A-A及6A-A-A单倍型高于对照组,差异有统计学意义(P＜0.05),而6A-G-G单倍型显著低于对照组(P＜0.01).结论 MMP-3血清水平在LAA型脑卒中急性期增高明显,SAO型脑卒中次之;中国北方汉族人群MMP-3基因rs3025058,rs522616及rs679620多态性可能与LAA型脑卒中易感性相关.     

难治性肾病综合征患儿载脂蛋白E基因多态性的研究

目的：研究难治性肾病综合征（steroid-resistant idiopathic nephrotic syndrome,SRINS)患儿载脂蛋白E基因多态性,为临床上正确选择合适的脂质代谢紊乱病例进行降脂治疗提供依据。方法：用酶法测定了60例SRINS患儿及80例健康儿童血脂、脂蛋白、载脂蛋白3种物质共7个脂质代谢指标,用PCR－SSCP法检测载脂蛋白E（apoE)基因型,并行肾穿刺活检术检查肾病综合征患儿病理类型。结果：SRINS患儿存在明显脂质紊乱,与健康儿童比较差异有显著性（P＜0．01）,随诊半年后仍有绝大多数SRINS患儿存在明显脂质代谢紊乱。难治性肾病综合征apoε2等位基因显著多于健康儿童（P＜0．05）。结论：SRINS患儿脂质代谢紊乱持续的时间较长,这类患儿,尤其携带ε2等位基因者,更易发生进行性肾脏损害,动脉粥样硬化及冠心病。应考虑给这类患儿使用降脂药物。     

The gene for apolipoprotein C-ll is closely linked to the gene for apolipoprotein E on chromosome 19

We have used a common TaqI restriction fragment length polymorphism (RFLP) near the human apolipoprotein C-II (apoC-II) gene to study linkage with apolipoprotein E (apoE). The inheritance of the apoC-II RFLP was followed in seven families that were segregating for apoE protein variants. No recombinants were observed in 20 informative meioses, giving an overall lod score of > 4.0 at recombination fraction 0. We have also observed apparent linkage disequilibrium between apoE and the apoC-II RFLP. Taken together these results demonstrate that these two apolipoprotein genes are closely linked and confirm that the gene for apoC-II is on human chromosome 19.     

血管性痴呆患者血浆中Tau蛋白与ApoE基因多态性的关系

目的:探讨血管性痴呆(VD)患者血浆中微管相关蛋白(Tau蛋白)与载脂蛋白E(ApoE)基因型的关系。方法:VD患者组102例,其中男65例,女37例,年龄60～72岁,平均年龄(65.8±5.8)岁。根据MMSE量表划分VD患者严重程度,分为轻度(20～24分)43例,中度(10～19分)38例,重度(10分以下)21例。选取100例健康老年者作为对照组,其中男58例,女42例,年龄58～75岁,平均年龄(64.2±6.7)岁。两组间年龄、性别构成无统计学差异。均排除其他神经系统及其他系统和物质原因所致的痴呆。所采集血标本置EDTA抗凝管中,低温离心取上清,加酸置低温冰箱保存。对所有人应用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)法进行ApoE基因多态性分析,同时应用ELISA法进行Tau蛋白定量分析。结果:经检测分析显示VD患者和ApoEε4等位基因者血浆中Tau蛋白升高,VD患者血浆中Tau蛋白浓度明显高于对照组。伴ApoEε4等位基因的VD患者与不伴ε4等位基因的VD患者相比,Tau蛋白浓度明显升高。结论:血浆中Tau蛋白浓度升高提示携带ε4等位基因VD患者早期易发生神经变性和神经纤维病理学改变;血浆中Tau蛋白浓度的检测可以作为早期VD诊断的生物学指标,尤其是携带ε4等位基因的VD患者。     

SHMT1基因启动子甲基化与缺血性卒中相关

目的探究丝氨酸羟甲基转移酶(SHMT1)基因甲基化与缺血性卒中的关系。方法采用甲基化特异性实时定量PCR测定290名健康对照组和141例缺血性卒中病例组(卒中组)的SHMT1甲基化水平。结果卒中组SHMT1甲基化水平为24.87%(16.97～35.46)高于对照组的6.58%(2.43～15.14)(P<0.05)。在调整相关危险因素后,SHMT1甲基化是卒中的危险因素(OR=1.051, 95%CI=1.034～1.068)。受试者工作特征曲线下面积为0.804,95%CI=0.760～0.849 (P<0.01)。在对照组发现尿酸与SHMT1甲基化相关(r_s=0.17,P<0.01),在卒中组发现三酰甘油与SHMT1甲基化相关(r_s=0.18,P<0.05)。SHMT1甲基化表达与mRNA的表达呈负相关(r=-0.472,P<0.01)。结论缺血性卒中患者中SHMT1基因启动子呈高甲基化状态,SHMT1低表达,且SHMT1高甲基化是卒中的危险因素。