Molecular epidemiology concerned with effect of nitric oxide synthase (NOS) gene polymorphism on NOS protein function in human cultured vascular endothelial cells

血管内皮细胞中一氧化氮合酶（eNOS）基因Glu298Asp多态性与心血管疾病的关系一直存在争议。为了验证eNOS基因Glu298Asp多态性对eNOS基因表达的影响，我们根据eNOS基因Glu298Asp多态性的基因型，利用培养的人血管内皮细胞检测eNOS mRNA和蛋白的表达。分娩后立即无菌取人脐带，经胶原酶处理收集人脐静脉内皮细胞（HUVECs），获得Glu298Glu或Glu298Asp基因型HUVECs。采用real-time PCR/RT-PCR检测eNOS mRNA表达。免疫分析法检测eNOS蛋白表达。含Glu298Asp的HUVECs的eNOS mRNA和蛋白表达倾向于高于含Glu298Glu的，但差异不显著。这些结果表明，Glu298Glu和Glu298Asp基因型差异对eNOS mRNA和蛋白的表达没有影响。

The relationship between cardiovascular disease and Glu298Asp polymorphism of the nitric oxide synthase (eNOS) gene in vascular endothelial cells is controversial due to conflicting results. To demonstrate the effects of Glu298Asp polymorphism of the eNOS gene on eNOS gene expression by excluding the effects of various confounding factors, eNOS mRNA and protein expressions were measured using cultured human vascular endothelial cells according to genotypes of Glu298Asp polymorphism of the eNOS gene. After human umbilical cords were aseptically obtained immediately after delivery, human umbilical vein endothelial cells (HUVECs) were collected by collagenase treatment, and HUVECs with the Glu298Glu or Glu298Asp genotype were obtained. eNOS mRNA expression was measured by real-time PCR/RT-PCR. eNOS protein expression was measured by an immunoassay method. HUVECs with Glu298Asp tended to show higher eNOS mRNA and protein expressions than those with Glu298Glu, but the difference was not significant. These results suggest that the genotypic difference of Glu298Glu and Glu298Asp has no effect on eNOS mRNA and protein expressions.