Effect of Interleukin‐10 and Laminar Shear Stress on African American Human Umbilical Vein Endothelial Cells

Abstract

African Americans (AA) have a predisposition to heightened systemic inflammation and a high prevalence of hypertension. The purpose of this study was to evaluate the influence of interleukin‐10 (IL‐10) and laminar shear stress (LSS) on AA endothelial cells by measuring total endothelial nitric oxide synthase (eNOS) protein expression and its activated, phosphorylated form (p‐eNOS) at Serine 1177, as well as nitric oxide (NO) levels, in response to IL‐10 incubation and high physiological levels of LSS, used as a mimetic for aerobic exercise training (AEXT). Human umbilical vein endothelial cells (HUVEC) from an AA donor were cultured. The experimental conditions included Static, Static with IL‐10 Incubation, LSS at 20 dynes/cm2, and LSS at 20 dynes/cm2 with IL‐10 Incubation. Western blotting was used to measure eNOS and p‐eNOS protein expression in the cells. A modified Griess assay was used to measure NO production in the cell culture media. There were significant increases in p‐eNOS, eNOS, and NO in the LSS at 20 dynes/cm2 and LSS at 20 dynes/cm2 with IL‐10 Incubation experimental conditions when compared to the Static experimental condition. There were no other statistically significant differences, demonstrating that IL‐10 did not have an additive effect on eNOS activity in the present study. The significant increases in p‐eNOS, eNOS, and NO as a result of LSS in AA HUVECs suggest that AEXT may be a viable, nonpharmacologic method to improve vascular inflammation status and vasodilation, and thereby contribute to reduction of hypertension in the AA population.AHA Scientist Development Grants 12SDG12070327NIH Grant RO1 [HL085497]