ESR measurement of endogenous nitric oxide in liver and blood of mice subjected to hepatic ischemia-reperfusion  Fusako Takayama, Toru Egashira, Yasumitsu Yamanaka  Pathophysiology  Volume 6, Issue 1, Pages 45-51 (April 1999) DOI: 10.1016/S0928-4680(98)00037-6

Fig. 1 ESR spectra detected at room temperature in mice liver homogenates. Subcutaneous injection of 0.3 M/kg (DTCS)/Fe was performed 1 h prior to liver resection. Treatment for mice was A, B, and C. A: (DTCS)/Fe injection alone, B: peritoneal administration of 50 mg/kg NMMA and then 2 h later (DTCS)/Fe injection, C: none. The characteristic three-line spectra of (DTCS)/Fe-NO adducts with the central g value of 2.040 and aN of 1.25 mT were detected in all samples from mice injected with (DTCS)/Fe. Pathophysiology 1999 6, 45-51DOI: (10.1016/S0928-4680(98)00037-6)

Fig. 2 ESR spectra of the (DTCS)/Fe-NO adducts in liver of mice at various reperfusion times following liver ischemia. Mice were subjected to 60 min of ischemia followed by reperfusion. A: a sham-operation; B: 60-min reperfusion; C: 6-h reperfusion; D: 12-h reperfusion. Subsequently, 1-h prior to liver resection, all mice received 0.3 M/kg (DTCS)/Fe subcutaneously. The 45 μl-aliquot of each 40% liver homogenate from the ischemic lobes was transferred into a capillary glass tube and put into a quartz for ESR measurement. Pathophysiology 1999 6, 45-51DOI: (10.1016/S0928-4680(98)00037-6)

Fig. 3 Change in the (DTCS)/Fe-NO concentrations in liver and blood samples after liver ischemia-reperfusion. Mice were subjected to 60 min of liver ischemia followed by the reperfusion denoted on the horizontal axis, or to sham-operation. Liver was divided into the right lateral and caudate lobes for the non-ischemic part (shaded square), and the left lateral and median lobes for the ischemic parts (filled square). Blood (shaded circle) was collected through the portal vein. Each value denotes accumulated amounts for 1 h, and represents the mean±S.E. for four mice. Pathophysiology 1999 6, 45-51DOI: (10.1016/S0928-4680(98)00037-6)

Fig. 4 Changes in blood endotoxin concentrations after ischemia-reperfusion. Mice were subjected to 60 min of ischemia followed by reperfusion. (Open circle): sham-operation group, (filled circle): liver ischemia-reperfusion group. Each value represents the mean±S.E. for four mice. *P<0.05, **P<0.01 vs sham operation group (Student’s t-test for unpaired data). Pathophysiology 1999 6, 45-51DOI: (10.1016/S0928-4680(98)00037-6)

Fig. 5 Liver PCOOH levels after liver ischemia-reperfusion. Mice were subjected to 60 min of ischemia followed by reperfusion denoted on the horizontal axis, or to sham-operation. (Open circle): sham-operation group; (open triangle): sham-operation group pretreated with NMMA; (filled circle): liver ischemia-reperfusion; (vertical lined triangle): liver ischemia-reperfusion group pretreated with NMMA. The PCOOH levels in liver tissues were determined by analyzing the lipid extracts from liver homogenates using CL-HPLC. Each value represents the mean±S.E. for four mice, with the treatment denoted on the horizontal axis. *P<0.05, **P<0.02 vs sham-operation group. #P<0.05 vs liver ischemia-reperfusion group without NMMA treatment. The significance of mean differences was analyzed by Student’s t-test for unpaired data. Pathophysiology 1999 6, 45-51DOI: (10.1016/S0928-4680(98)00037-6)