内皮型NO合酶基因多态性与冠心病、吸烟的关系

A Smoking-dependent Risk of Coronary Artery Disease Associated With A Polymorphism of The Endothelial Nitric Oxide Synthase Gene

目的探讨内皮型一氧化氮合酶(endothelialnitricoxidesynthase,eNOS)基因多态性与冠心病、吸烟的关系。方法依据eNOS基因外显子7G894T位点设计引物,通过巢式聚合酶链反应(PCR)扩增目的片段,限制性内切酶消化目的片段,琼脂糖凝胶电泳,紫外透射分析仪检测,计数117例CAD患者和有吸烟的CAD患者及100例健康者基因型及突变基因频率,通过χ2检验有无统计学意义。结果eNO基因外显子7的894位点有3种基因型:GG、GT、TT。CAD组117例中31例发生G894T突变,纯合子TT3例,杂合子GT28例。对照组100例中15例发生G894T突变,均为杂合子。CAD组基因突变频数明显高于对照组,两组GT+TT型χ2检验结果:χ2=4.265P<0.05,等位基因频数χ2=5.321,P<0.05。吸烟比例在CAD组明显高于对照组χ2=17.921,P<0.01。另外,基因突变频数在CAD吸烟组明显高于CAD非吸烟组χ2=4.966,P<0.05。结论eNOS基因894位点G→T突变与吸烟和CAD发病密切相关。

Aim To investigate the relationship between the gene polymorphism of the endothelial nitric oxide synthase (eNOS) and coronary artery disease (CAD) concerning with smoking. Mthods By using the designed primers based on the 7G894T sites of the eNOS exon. The target fragments were amplified by nested PCR and digested by restriction enzyme, then the agaroae gel electroghoresis and ultraviolet transmitting analysis were used for detection. By counting the genotype and mutation frequencies of 117 patients with or without smoking habit as well as 100 healthy people, the statistic significance was assessed with chi square test. Rsults three genotypes were identified as GG, GT, and TT on the 894 site of the eNOS gene exon 7. In CAD group, 31 out of 137 patients were identified as 7G894T mutation including 3 homozygotes and 28 heterozygotes .In control group 15 out of 100 patients had G894T mutation, who are all heterozygotes. Frequency of gene mutation in CAD group were significantly higher than the control group, the results of chi square test for the two groups of GT and TT were as follows:χ 2=4.265, P< 0.05,The frequency of allele χ 2= 5.321 P< 0.05, the frequency of smoking in CAD group were significant higher than the control group χ 2= 17.921, (P< 0.01). In addition, the frequency of gene mutation in smoking CAD group was significantly higher than nonsmoking one.χ 2=4.966, P< 0.05. <P>Cnclusion The G→ T mutation of G894T site in eNOS gene is associated with CAD concerning with smoking.<P>