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

血管内皮细胞一氧化氮合酶（eNOS）基因Glu 298 Asp多态性与心血管疾病的关系存在争议。为了排除各种混杂因素的影响，证明eNOS基因的Glu 298 Asp多态性对eNOS基因表达的影响，根据eNOS基因的Glu 298 Asp多态性的基因型，使用培养的人血管内皮细胞测量eNOS mRNA和蛋白质表达。在无菌条件下获得人脐带后，立即通过胶原酶处理收集人脐静脉内皮细胞（HUVECs），并获得具有Glu 298 Glu或Glu 298 Asp基因型的HUVECs。实时定量PCR/RT-PCR检测eNOS mRNA表达。用免疫分析法测定eNOS蛋白表达。Glu 298 Asp转染的HUVECs eNOS mRNA和蛋白表达均高于Glu 298 Glu转染的HUVECs，但差异无统计学意义。提示Glu 298 Glu和Glu 298 Asp基因型差异对eNOS mRNA和蛋白表达无影响。

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.