EPR analysis of extra‐ and intracellular nitric oxide in liver biopsies

Purpose To develop an assay that can enable the quantification of intra‐ and extracellular nitric oxide (NO) levels in liver biopsies without application of potentially harmful exogenous NO traps. Theory Electron paramagnetic resonance (EPR) spectroscopy is currently the most appropriate method of m...

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Bibliographic Details
Published in:Magnetic resonance in medicine 2017-06, Vol.77 (6), p.2372-2380
Main Authors: Dumitrescu, Sergiu D., Meszaros, Andras T., Puchner, Stefan, Weidinger, Adelheid, Boros, Mihaly, Redl, Heinz, Kozlov, Andrey V.
Format: Article
Language:English
Subjects:
Animals
Biological samples
Biomarkers - analysis
Biopsy
Blood levels
Cells, Cultured
dinitrosyl–iron complex
Electron paramagnetic resonance
Electron Spin Resonance Spectroscopy - methods
endogenous NO traps
Extracellular Fluid - chemistry
Hemoglobin
Hepatitis - metabolism
Inflammation
Intracellular Fluid - chemistry
Iron
Lipopolysaccharides
Liver
Liver - chemistry
Liver - pathology
Male
mononitrosyl‐hemoglobin complex
Nitric oxide
Nitric Oxide - analysis
Rats
Rats, Sprague-Dawley
Reproducibility of Results
Resonance
Sensitivity and Specificity
Spectroscopy
Traps
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Summary:Purpose To develop an assay that can enable the quantification of intra‐ and extracellular nitric oxide (NO) levels in liver biopsies without application of potentially harmful exogenous NO traps. Theory Electron paramagnetic resonance (EPR) spectroscopy is currently the most appropriate method of measuring NO in biological samples due to the outstanding specificity resulting from the interaction of NO with exogenous NO traps. Because such traps are not allowed in clinical settings, we tested the reliability of endogenous NO traps for the determination of NO levels in blood and liver compartments. Methods Rats were injected with 0–8 mg/kg lipopolysaccharide (LPS) to gradually induce a systemic inflammatory response. Specific features of NO‐hemoglobin and NO–Fe EPR signals were quantified using a specifically developed calibration procedure. Results Whereas both NO–hemoglobin (NO–HbLIVER BLOOD) and NO–Fe (NO–FeLIVER) complexes were detected in nonperfused liver tissue, only NO–Fe complexes were detected in perfused tissue and only NO–Hb complexes were detected in blood (NO–HbBLOOD). The NO concentrations increased in the sequence NO–HbBLOOD 
ISSN:0740-3194
1522-2594
DOI:10.1002/mrm.26291