Effect of Interleukin-10 and Laminar Shear Stress on Endothelial Nitric Oxide Synthase and Nitric Oxide in African American Human Umbilical Vein Endothelial Cells

Abstract

<p><strong>Background: </strong>African Americans have a pre­disposition to heightened systemic inflamma­tion and a high prevalence of hypertension.</p><p><strong>Objective: </strong>The purpose of this study was to evaluate the influence of interleukin-10 (IL- 10) and laminar shear stress (LSS) on African American endothelial cells by measuring to­tal endothelial nitric oxide synthase (eNOS) protein expression and its phosphorylated form (p-eNOS) at Serine 1177, and nitric oxide (NO) levels, in response to IL-10 incubation and high physiological levels of LSS, used as an <em>in vitro </em>mimetic for aerobic exercise training (AEXT).</p><p><strong>Design: </strong>Human umbilical vein endothelial cells (HUVEC) from an African American donor were cultured. The experimental conditions included <em>Static</em>, <em>Static with IL-10 Incubation, LSS at 20 dynes/cm</em><em>2</em><em>, and LSS at 20 dynes/cm</em><em>2 </em><em>with IL-10 Incubation</em>. West­ern blotting was used to measure eNOS and p-eNOS protein expression in the cells. A modified Griess assay was used to measure NO metabolites in the cell culture media.</p><p><strong>Results: </strong>There were significant increases in p-eNOS, eNOS, and NO in the <em>LSS at 20 dynes/cm</em><em>2 </em>and <em>LSS at 20 dynes/cm</em><em>2 </em><em>with IL-10 Incubation </em>experimental conditions when compared to the <em>Static </em>experimental condition. There were no other statistically significant differences demonstrating that IL- 10 did not have an additive effect on eNOS activity in our study.</p><p><strong>Conclusion: </strong>The significant increases in p-eNOS, eNOS, and NO as a result of LSS in African American HUVECs suggest that AEXT may be a viable, nonpharmacologic method to improve vascular inflamma­tion status and vasodilation, and thereby contribute to hypertension reduction in the African American population. <em>Ethn Dis. </em>2015;25(4):413-418; doi:10.18865/ ed.25.4.413</p>

Endothelial Dysfunction in Chronic Kidney Disease, from Biology to Clinical Outcomes: A 2020 Update

“…Chronic exposure to increased shear stress enhances eNOS activity through upregulation of the phosphorylation of this enzyme, resulting in higher production of NO. This mechanism might explain the beneficial effects of aerobic exercise training on endothelial function [40,45].…”

“…Increased endothelial shear stress induced by an increase in blood flow may enhance the activity of eNOS in a calcium-independent way [38,39]. In human coronary circulation, an acute increase of shear stress promotes the generation of BK, which subsequently upregulates the function of eNOS via a Gq-dependent pathway [11,39,40]. Moreover, immediate exposure to shear forces may increase the permeability of human glomerular endothelial cells through upregulation of eNOS activity [41].…”

Irisin Lowers Blood Pressure by Improvement of Endothelial Dysfunction via AMPK‐Akt‐eNOS‐NO Pathway in the Spontaneously Hypertensive Rat

“…Concentrations of NO metabolites nitrite and nitrate in the cell‐culture supernatant were determined using an assay based on the enzymatic conversion of nitrate to nitrite by nitrate reductase, followed by colorimetric detection of nitrite as an azo‐dye product of the Griess reaction (R&D Systems; Minneapolis, MN). All samples were centrifuged to remove particulates at 16 000 g for 20 minutes at 4°C . One hundred microliters of each supernatant were mixed with 100 μL of the Griess reagent for 10 minutes at 37°C, and absorbance was recorded on a 96‐well plate using Thermo Scientific Varioskan Flash (Thermo LabSystems, Inc., Philadelphia, PA) at 540 nm .…”

“…All samples were centrifuged to remove particulates at 16 000g for 20 minutes at 4°C. 33 One hundred microliters of each supernatant were mixed with 100 lL of the Griess reagent for 10 minutes at 37°C, and absorbance was recorded on a 96-well plate using Thermo Scientific Varioskan Flash (Thermo LabSystems, Inc., Philadelphia, PA) at 540 nm. 34 Total nitrite levels were determined using a standard curve.…”

TNF gene polymorphism as a risk factor that can cause arterial hypertension in patients suffering from gastroesophageal reflux disease

“…Помимо традиционных факторов риска возникновения и прогрессирования АГ и ГЭРБ, активно изучается патогенетическая роль медиаторов воспаления, обсуждается и ассоциация провоспалительных цитокинов с развитием эндотелиальной дисфункции и увеличением жесткости сосудистой стенки [6,7].…”