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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。
Objective To investigate the relationship between endothelial nitric oxide synthase (eNOS) gene polymorphism and coronary heart disease and smoking. Methods According to the site of exon 7G894T of eNOS gene, primers were amplified by Nested Polymerase Chain Reaction (PCR), digested with restriction endonuclease and analyzed by agarose gel electrophoresis and ultraviolet transmission analyzer The genotype and mutation frequency of 117 CAD patients, smoking CAD patients and 100 healthy controls were all statistically significant by χ2 test. Results There were three genotypes at the 894 site in exon 7 of eNO gene: GG, GT and TT. In the CAD group, G894T mutation occurred in 31 cases, homozygous TT3 cases and heterozygous GT28 cases. In control group, 15 cases of G894T mutation occurred in 100 cases, all of which were heterozygous. The frequencies of gene mutation in CAD group were significantly higher than those in control group. The results of χ2 = 4.265P <0.05 and the allele frequency χ2 = 5.321, P <0.05. Smoking ratio in CAD group was significantly higher than the control group χ2 = 17.921, P <0.01. In addition, the frequency of gene mutation in CAD smoking group was significantly higher than CAD non-smoking group χ2 = 4.966, P <0.05. Conclusion The G → T mutation at 894 site of eNOS gene is closely related to the incidence of smoking and CAD.