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Magnetic resonance study of the transmembrane nitrite diffusion.

A Samouilov¹, Ya Yu Woldman, J L Zweier

¹Center for Biomedical EPR Spectroscopy and Imaging, The Davis Heart and Lung Research Institute, The Department of Internal Medicine, The Ohio State University College of Medicine, Columbus, OH 43210, USA. Alex.Samouilov@osumc.edu

Nitric Oxide : Biology and Chemistry

|February 20, 2007

Summary

Nitrite (NO2-) enters cells as nitrous acid (HNO2), causing internal acidification. This mechanism explains nitrite accumulation and its role in tissue injury, impacting nitric oxide (NOx) levels.

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In Situ Monitoring of Diffusion of Guest Molecules in Porous Media Using Electron Paramagnetic Resonance Imaging

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Area of Science:

Biochemistry
Cell Biology
Physiology

Background:

Nitrite (NO2-) is a metabolite of nitric oxide (NO*) and nitrate (NO3-).
Nitrite can accumulate in tissues and regenerate NO, with implications for ischemia/reperfusion injury.
Mechanisms of intracellular nitrite accumulation are not well understood.

Purpose of the Study:

To investigate the role of undissociated nitrous acid (HNO2) in nitrite (NO2-) penetration through biological membranes.
To elucidate the mechanisms of intracellular nitrite accumulation.
To assess the physiological significance of nitrite transport and its effects on tissue NOx levels.

Main Methods:

Utilized large unilamellar phosphatidylcholine liposomes and spectroscopic techniques (EPR, 31P NMR) to study nitrite transport.
Employed pH-sensitive probes and chemiluminescence to measure intraliposomal acidification and nitrite accumulation.
Perfusion of isolated rat hearts with nitrite-containing buffer to assess tissue penetration and NOx levels.

Main Results:

Demonstrated that nitrous acid (HNO2) transport across liposomal membranes causes intraliposomal acidification.
Confirmed nitrite (NO2-) accumulation within liposomes against a concentration gradient when an alkaline pH gradient was applied.
Observed rapid nitrite penetration into isolated rat hearts, leading to a significant increase in myocardial NOx concentration.

Conclusions:

Nitrite (NO2-) passively penetrates biological membranes as undissociated nitrous acid (HNO2).
This transport is coupled with proton transfer, leading to intraliposomal acidification.
The findings provide a mechanism for intracellular nitrite accumulation and its potential physiological roles in conditions like ischemia/reperfusion injury.