A Comparative ESR Study on Blood and Tissue Nitric Oxide Concentration during Renal Ischemia-Reperfusion Injury |
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| Authors: | J Ren P C W Fung C Chang G X Shen G Lu F H Y Chan K J Liu J Shen |
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| Institution: | (1) Department of Electrical and Electronic Engineering, University of Hong Kong, Hong Kong, China;(2) Department of Medicine, University of Hong Kong, Hong Kong, China;(3) Fu Tian Hospital, Guangdong Medical College, Shenzhen, China;(4) College of Pharmacy, University of New Mexico, Albuquerque, New Mexico, USA;(5) School of Chinese Medicine, University of Hong Kong, Hong Kong, China |
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| Abstract: | Electron paramagnetic resonance (EPR) spin trapping technology is a sensitive and unambiguous method for detection of nitric
oxide (NO). Due to the short lifetime, NO must be trapped before EPR measurement. There are two EPR spin trapping techniques
used currently, including the detections of EPR signals of diethyldithiocarbamate-iron-nitric oxide (DETC2-Fe2+-NO) and nitrosyl hemoglobin (HbNO). In this study, we firstly investigated the kinetics of the EPR signal of DETC2-Fe2+-NO in normal and ischemia-reperfused kidneys. In normal rat kidneys, the signal of DETC2-Fe2+-NO was found at 5 min after the spin trappers Fe2+/DETC were administrated, the peak concentration was at 15 min and the period with relatively stable signal intensity was
at the time range from 15 to 70 min. In the ischemia-reperfused rat kidneys, the signal of DETC2-Fe2+-NO was increased at 30 min of ischemia and decreased at 60 min of ischemia after the occlusion of renal artery (corresponding
to the time course of 60 and 90 min after Fe2+/DETC injection respectively). We then investigated the EPR signal of HbNO in blood. No characteristic HbNO signal was found
in the rats of the sham control and 30 min of ischemia. An HbNO signal occurred in the rats exposed to 60 min of ischemia
and it became pronounced with increased duration of reperfusion. The signal intensity reached a plateau at 150 min of reperfusion.
The results suggest that the DETC2-Fe2+-NO signal can be only suitable for the NO measurement in the short-term ischemia-reperfusion model, whereas the HbNO signal
can be applied to represent NO in the relatively long-term ischemia-reperfusion model. In addition, NG-nitro-L-arginine (L-NAME) and allopurinol were used to identify the source of NO. By detecting the HbNO signal, we demonstrated
that the activation of xanthine oxidase is an important source of NO formation at the long-term period of ischemia and reperfusion.
Authors' address: Jiangang Shen, School of Chinese Medicine, University of Hong Kong, 10 Sassoon Road, Hong Kong SAR, China |
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