Abstract

The broad, long-term goal of this project is to investigate the diffusion and reaction of nitric oxide (NO) in blood, particularly its interaction with red blood cells (RBCs). It is hypothesized that RBCs possess specific mechanisms that regulate the NO consumption rate through modulation of membrane permeability to NO. Specifically the following aims will be pursued.
Specific Aim 1 : Is NO consumption by RBC regulated by transmembrane diffusion? Specific Aim 2: Do any specific intra- erythrocytic molecules participate in the regulation of NO quenching? Specific Aim 3: How does the regulation of NO consumption by RBCs affect vessel regulation? The first two aims will be addressed by use of a competitive experiment and a differential membrane bioreactor specifically designed to measure the NO-RBC reaction rate. Kinetic models will be used to analyze the data. Biophysical (EPR and fluorescence) and biochemical (characterization of enzymes, metabolites, and lipids) techniques will be applied to RBCs, RBC ghosts, and synthetic liposomes in order to answer these questions. The last aim will be addressed using isolated porcine coronary microvessels as a bio-assay to determine the functional role of NO quenching and its regulation. The hypotheses proposed above are a significant departure from the current understanding that NO consumption is not regulated and that the RBC membrane is """"""""completely permeable"""""""" to NO. In addition to its contribution to fundamental physiology, the proposed work directly impacts multiple aspects of clinical medicine, including NO inhalation therapy and the design of blood substitutes. Furthermore, the proposed mechanism might contribute to the pathology of several diseases, such as essential and pulmonary hypertension, peripheral vascular disease associated with diabetes mellitus, sickle cell anemia, and other hereditary RBC disorders. In these situations, altered RBC membrane consumption by RBCs is essential to the development of clinical intervention and understanding of the complex roles that NO plays under physiological and pathological conditions.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL065741-03
Application #
6527652
Study Section
Surgery and Bioengineering Study Section (SB)
Program Officer
Qasba, Pankaj
Project Start
2000-09-27
Project End
2004-07-31
Budget Start
2002-08-01
Budget End
2003-07-31
Support Year
3
Fiscal Year
2002
Total Cost
$293,250
Indirect Cost

  Institution

Name
University of California Los Angeles
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095

  Publications

Wang, Wei; Hein, Travis W; Zhang, Cuihua et al. (2011) Oxidized low-density lipoprotein inhibits nitric oxide-mediated coronary arteriolar dilation by up-regulating endothelial arginase I. Microcirculation 18:36-45
Chou, Katherine J; Dodd, Joanna; Liao, James C (2008) Interactions of nitrosylhemoglobin and carboxyhemoglobin with erythrocyte. Nitric Oxide 18:122-35
Hyduke, Daniel R; Jarboe, Laura R; Tran, Linh M et al. (2007) Integrated network analysis identifies nitric oxide response networks and dihydroxyacid dehydratase as a crucial target in Escherichia coli. Proc Natl Acad Sci U S A 104:8484-9
Han, Tae H; Pelling, Andrew; Jeon, Tae-Joon et al. (2005) Erythrocyte nitric oxide transport reduced by a submembrane cytoskeletal barrier. Biochim Biophys Acta 1723:135-42
Hyduke, Daniel R; Liao, James C (2005) Analysis of nitric oxide donor effectiveness in resistance vessels. Am J Physiol Heart Circ Physiol 288:H2390-9
Han, Tae H; Fukuto, Jon M; Liao, James C (2004) Reductive nitrosylation and S-nitrosation of hemoglobin in inhomogeneous nitric oxide solutions. Nitric Oxide 10:74-82
Han, Tae H; Qamirani, Erion; Nelson, Allyson G et al. (2003) Regulation of nitric oxide consumption by hypoxic red blood cells. Proc Natl Acad Sci U S A 100:12504-9
Han, Tae H; Hyduke, Daniel R; Vaughn, Mark W et al. (2002) Nitric oxide reaction with red blood cells and hemoglobin under heterogeneous conditions. Proc Natl Acad Sci U S A 99:7763-8
Joshi, Mahesh S; Ferguson Jr, T Bruce; Han, Tae H et al. (2002) Nitric oxide is consumed, rather than conserved, by reaction with oxyhemoglobin under physiological conditions. Proc Natl Acad Sci U S A 99:10341-6
Huang, K T; Han, T H; Hyduke, D R et al. (2001) Modulation of nitric oxide bioavailability by erythrocytes. Proc Natl Acad Sci U S A 98:11771-6