DETECTION OF NITRIC OXIDE REDUCTION DURING ISCHAEMIA-REPERFUSION BY EPR SPECTROSCOPY

Abstract

Introduction: Acute renal failure is a common consequence of sepisis due to concurrent renal ischaemia. The role of nitric oxide (NO) in endotoxaemia and in ischaemic injury in the kidney is not well defined. Material and Methods: In this study we have used an animal model of sepsis induced by injection of bacterial lipopolysaccharide (LPS) in the rat and measured renal nitric oxide by X-band electron paramagnetic resonance (EPR) spectroscopy using the spin trap Fe2+-N-methyl-D-glucamine dithiocarbamate [Fe(MGD)2] given by intravenous injection 6 minutes before sacrifice. Results: The characteristic EPR spectrum of [Fe(NO)(MGD)2] was observed in kidneys of rats treated with LPS for 5h. Rat kidneys subjected to 20 min ischaemia and 5 min reperfusion had lower concentrations of [Fe(NO)(MGD)2] (1.0 ± 0.6 (M) compared to the contralateral nonischaemic kidneys (1.5 ± 0.9 (M, P<0.05). Conclusion: This study shows reduced levels of NO after renal ischaemia in vivo.

Keywords

• Nitric oxide
• Electron paramagnetic resonance
• Kidney
• Ischaemia. reperfusion