冠心病患者vWF基因A1381T多态性分析

Analysis of vWF Gene A1381T Polymorphism in Patients with Coronary Heart Disease

本研究探讨冠心病患者vWF基因A1381T多态性。采用酶联免疫吸附法测定104名40-75岁(平均59岁)连续住院的冠心病患者血浆vWF:Ag水平,同时测定96名39-70岁(平均56岁)同期门诊体检的人群血浆vWF:Ag水平作为对照组;采用PCR产物酶切片段长度分析vWF基因A1381T单核苷酸多态性,测序验证。实验数据根据对照组和冠心病组性别、血型或/和基因型进行分组,采用t检验、方差分析、χ2检验等统计学处理。结果表明:vWF基因A1381T多态性GG基因型的频率在冠心病组为62.5%,对照组为67.7%,而AG基因型在冠心病组为37.5%,对照组为32.3%。卡方检验显示,AG基因型与冠心病没有相关性,其优势比OR=1.258(95%CI=0.702-2.255,χ2=0.595,p=0.440)。冠心病组血浆vWF含量明显高于对照组(p<0.001),冠心病组AG和GG基因型的个体血浆vWF含量均明显高于对照组AG和GG型(p<0.001)。基因型AG与GG比较,在冠心病组中差异有统计学意义(p<0.05),即冠心病组AG个体血浆vWF含量明显高于GG个体,而对照组AG个体血浆vWF含量虽略有增高但无统计学意义(p>0.05)。在对照组中,O型与A、B和AB型比较其血浆vWF含量明显降低(p<0.05);而A、B及AB型之间血浆vWF含量差异没有统计学意义(p>0.05)。在冠心病组中,O、A、B和AB型之间相互比较血浆vWF含量之间均没有差异(p>0.05);经过两组相同血型的比较,冠心病组的B、AB、O血型对应的血浆vWF含量均高于对照组(p=0.000、0.007和0.000),而A血型的血浆vWF含量也高于对照组,但没有统计学意义(p=0.06)。双因素方差分析结果表明,血型和A1381T基因多态性对血浆vWF水平的影响没有交互作用,但冠心病组O血型的AG基因型个体与GG基因型个体相比,血浆vWF含量明显增高(p<0.05),其他血型AG基因型个体与GG基因型个体相比,血浆vWF含量没有明显差异(p>0.05)。结论 :vWF基因A1381T多态性与冠心病的易感性没有相关性,冠心病组血浆vWF水平随ABO血型和vWF基因A1381T多态性不同而有明显的不同,尤其AG基因型血浆vWF增高最明显,冠心病组O血型A1381T多态性的AG基因型血浆vWF水平明显高于GG基因型。这些结果对于进一步研究A1381T多态性是否影响vWF基因表达及vWF活性具有一定的参考意义。

This study was purposed to investigate the vWF gene A1381T polymorphism in patients with coronary heart disease （CHD）. A case-control study was designed, including 104 continuously hospitalized patients with CHD, aging from 40 to 75 years （ average 59） and 96 persons underwent physical examination in outpatient department as controls, aging from 39 to 70 years （ average 56）. The plasma vWF： Ag level of CHD patients and control persons was detected by ILISA. vWF gene A1381 T polymorphism was analyzed by the polymerase chain reaction-restriction fragment length polymorphism and sequencing when it is necessary. The data were grouped by gender, blood group and/or genotype in CHD group and control groups. The difference of plasma vWF level between male and female was analyzed by independent sample t test; one way ANOVA was used to analyze the difference of vWF level between different blood group genotypes, while the factorial design ANOVA was used to test the difference of vWF level in plasma between A1381T genotype and/or ABO blood groups. χ2 Crosstabs were used to test the CHD susceptibility. The results showed that the frequencies of GG genotype （wild type） of vWF gene A1381T polymorphism were 62.5% in CHD group and 67.7% in control group, and the frequencies of AG genotype （heterozygous variant） were 37.5% in CHD group and 32.3% in control group. χ2 Crosstabs showed no significant correlation between vWF gene A1381T polymorphism （AG） and CHD （ OR = 1. 258, 95% CI = 0. 702 - 2. 255, χ2 = 0. 595 ,p = 0. 440）. The plasma vWF level in CHD group was statistically very higher than that in control group（p 〈 0. 001 ）, even though the relationship of vWF A1381T polymorphism （rs216311 ） and susceptibility of CHD in CHD group was not found. The plasma vWF level of AG or GG genotype was higher in CHD group than in control group（p 〈 0. 001 ）. The plasma vWF level of AG genotype was higher than that of GG in CHD group （p 〈 0.05 ）, but not in control group. The plasma vWF of O blood group was lower than that of A, B and AB blood groups（p 〈0.05）, while among A, B, AB blood groups, the vWF level was not different （p 〉 0.05 ）. Among O, A, B, AB blood groups in CHD group , vWF level was not different （p 〉 0.05 ）. Although the two-way analysis of variance ANOVA showed no interaction of A1381T genotype and ABO blood groups on plasma vWF level, the plasma vWF level in AG mutant of vWF A1381T gene polymorphism with O blood group was higher than that of GG mutant（p =0.023 ） in CHD group, not different in other blood groups. It is concluded that there is no association between vWF gene A1381 T polymorphism and CHD susceptibility. The plasma vWF level in CHD group interrelated with ABO blood group and A1381T polymorphism, in which the plasma vWF level in AG genotype increase mostly. Plasma vWF level in vWF gene A1381 T polymorphism with AG mutant was significantly much higher than GG mutant in CHD. This change may be beneficial to further study the effect of A1381 T polymorphism on vWF gene expression and activity.