TY - JOUR
T1 - Direct detection of tissue nitric oxide in septic mice
AU - James, Philip E.
AU - Liu, Ke J.
AU - Swartz, Harold M.
PY - 1998
Y1 - 1998
N2 - Although nitric oxide (NO) is a central mediator during endotoxin- induced sepsis, direct detection of tissue NO in vivo, has until recently been difficult, and techniques have relied on indirect measurement of bi- products in blood or invasive technology. We have utilized electron paramagnetic resonance (EPR) in conjunction with the spin-trapping technique to detect NO directly, and non-invasively, from the tissue of septic mice. Relative signal intensity arising from NO complexed with iron and diethyldithiocarbamate (DETC) measured directly from the liver and kidney of mice given endotoxin was maximal at 6 hours post endotoxin. We failed to detect an EPR signal from mice given pyrogen-free saline. The quality of the EPR signal obtained (high signal to noise ratio of 15:1) using our experimental set-up and L-band EPR hardware was such that we were able to establish a time course of NO production in tissue following endotoxin, and measurement of NO from other organs (kidney and spleen). Our EPR results probably reflected NO arising from inducible NO-synthase enzymes as a result of endotoxin stimulation. This technique was extended to experiments in which we first implanted an oxygen sensitive material (gloxy) into the liver of mice, and then monitored NO production following endotoxin. Due to the fact that the EPR spectrum from gloxy and that of NO-Fe(DETC)2 do not overlap, we were able to monitor NO production and pO2 simultaneously in tissue, in real time.
AB - Although nitric oxide (NO) is a central mediator during endotoxin- induced sepsis, direct detection of tissue NO in vivo, has until recently been difficult, and techniques have relied on indirect measurement of bi- products in blood or invasive technology. We have utilized electron paramagnetic resonance (EPR) in conjunction with the spin-trapping technique to detect NO directly, and non-invasively, from the tissue of septic mice. Relative signal intensity arising from NO complexed with iron and diethyldithiocarbamate (DETC) measured directly from the liver and kidney of mice given endotoxin was maximal at 6 hours post endotoxin. We failed to detect an EPR signal from mice given pyrogen-free saline. The quality of the EPR signal obtained (high signal to noise ratio of 15:1) using our experimental set-up and L-band EPR hardware was such that we were able to establish a time course of NO production in tissue following endotoxin, and measurement of NO from other organs (kidney and spleen). Our EPR results probably reflected NO arising from inducible NO-synthase enzymes as a result of endotoxin stimulation. This technique was extended to experiments in which we first implanted an oxygen sensitive material (gloxy) into the liver of mice, and then monitored NO production following endotoxin. Due to the fact that the EPR spectrum from gloxy and that of NO-Fe(DETC)2 do not overlap, we were able to monitor NO production and pO2 simultaneously in tissue, in real time.
UR - http://www.scopus.com/inward/record.url?scp=0032439805&partnerID=8YFLogxK
U2 - 10.1007/978-1-4615-4863-8_22
DO - 10.1007/978-1-4615-4863-8_22
M3 - Article
C2 - 9889891
AN - SCOPUS:0032439805
SN - 0065-2598
VL - 454
SP - 181
EP - 187
JO - Advances in Experimental Medicine and Biology
JF - Advances in Experimental Medicine and Biology
ER -