Analysis of the expression of eNOS in erythrocytes under hypoxic conditions (1013.15)

Abstract

Background: Congenital heart diseases are among the most common birth defects, being the second leading cause of infant death in Mexico. Some of these pathologies are characterized by low concentrations of O2 in the blood, resulting in cyanosis. There is evidence that erythrocytes express active and functional eNOS, which catalyzes the production of nitric oxide (NO). NO activity in erythrocytes appears to include the regulation of membrane deformability and inhibition of platelet activation; however, the activities of eNOS, NO export to the vasculature and the relative contribution of erythrocytes to the active NO pool in circulation, have not yet been explored in detail.Objective: To identify differences in eNOS expression and activity in erythrocytes from children with congenital heart disease and cyanosis, compared to healthy controls.Methods: Analysis of eNOS expression and activity by western blot and fluorometry, in erythrocytes of patients diagnosed with congenital heart disease and blood O2 saturation below 80%, at the Children's Hospital‐IMIEM, Toluca, Mexico.Results: 30 samples from patients with congenital heart disease and blood O2 saturation less than 80%, and 6 samples from healthy controls matched for age and sex, have been collected. Erythrocytes of all samples have been isolated and changes in eNOS expression are being analyzed. Enzymatic activity is being analyzed by subjecting the erythrocytes to mechanical stress and by measuring changes in intracellular calcium concentrations and NO production.