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

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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.