Neutrophil accumulation in ischemic reperfused rat liver: evidence for a role for superoxide free radicals.
Oxygen-derived free radicals and leukocytes have been implicated in the pathogenesis of ischemia-reperfusion injury. This study aimed at determining, by using biochemical and histochemical techniques, whether an accumulation of neutrophils occurs in the ischemic reperfused rat liver and whether superoxide free radicals play a role in mediating this neutrophil accumulation. Hepatic ischemia was induced by occluding blood supply to the left and median lobes, and reperfusion was reinstituted by releasing the occlusion. Myeloperoxidase activity of the liver was measured with a tetramethylbenzidine-H2O2 assay after removal of glutathione (by dialysis) and in the presence of 3-aminotriazole (catalase inhibitor). A modification of Graham and Karnovsky's method was used to stain neutrophils in liver frozen sections, and the number of neutrophils was counted. Results showed that ischemia-reperfusion of the liver produced a 4.4-fold increase in myeloperoxidase activity (from 0.073 +/- 0.009 to 0.320 +/- 0.017 units/mg liver, means +/- SE), which was proportional to the number of neutrophils (3.1-fold increase from 18 +/- 7 to 57 +/- 4 cells/mm2) in the liver tissue. Pretreatment with long-acting superoxide dismutase significantly attenuated the elevated myeloperoxidase activity and the number of neutrophils. These results indicate that reperfusion after a period of ischemia induces an accumulation of neutrophils in the liver, and superoxide anion free radicals are important mediators in the mechanism of this neutrophil accumulation.
Komatsu
H
H
Department of Medicine, University of Southern California, School of Medicine, Los Angeles 90033.
Koo
A
A
Ghadishah
E
E
Zeng
H
H
Kuhlenkamp
J F
JF
Inoue
M
M
Guth
P H
PH
Kaplowitz
N
N
D000818:Animals; D005609:Free Radicals; D007511:Ischemia / Q000201:enzymology / Q000473:pathology*; D007958:Leukocyte Count; D008099:Liver / Q000201:enzymology; D008102:Liver Circulation; D008297:Male; D009504:Neutrophils / Q000201:enzymology / Q000473:pathology*; D009195:Peroxidase / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D015424:Reperfusion; D013194:Staining and Labeling; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism*
D005609:Free Radicals; D013481:Superoxides; D009195:Peroxidase; D013482:Superoxide Dismutase