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Endothelial modulation of nitric oxide synthase by the no donor (RuBPY)

Grant number: 16/25300-5
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): April 01, 2017
Effective date (End): March 31, 2018
Field of knowledge:Biological Sciences - Pharmacology - Cardiorenal Pharmacology
Principal researcher:Lusiane Maria Bendhack
Grantee:Jessica Tamiris Romano
Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil


The endothelial cells produce and release relaxing dependent endothelial factors (EDRFs) and contractile factors. The major EDRF is nitric oxide (NO) that is produced by endothelial nitric oxide synthase (eNOS). Uncoupled eNO produces oxygen reactive species (ROS). eNOS can be expressed as dimer (active enzyme) or monomer (inactive enzyme). NO donors can release NO in the endothelial cells and in the vascular smooth muscle cells (VSMC). The hypothesis of this study is that the NO donor (RuBPY) positively modulates eNOS activity, increasing NO production in normotensive rat vessels that do not present endothelial dysfunction. Therefore, this project aims to study the relaxation induced by RuBPY in intact endothelium rat aorta in order to evaluate the positive modulation of this compound on eNOS activity. In rat isolated aortas, we will study the vascular reactivity, flow citometry and Western Blot. The responses to RuBPY will be evaluated in the presence of eNOS inhibitor (L-NAME), its substrate (L-Arginine) and superoxide scavenger (Tiron). We will study the relaxation induced by acetylcholine and its modulation by RuBPY. By using flow citometry, we will measure the cytosolic NO concentration ([NO]c) in the human umbilical vein endothelial cells (HUVECs) by the NO sensitive dye DAF-2DA and the ROS production by DHE. By using Western Blot, we will verify if the enzyme eNOS is active (dimer) or if it is uncoupled and inactive (monomer). (AU)

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