Superoxide generation by neutrophils and Kupffer cells during in vivo reperfusion after hepatic ischemia in rats.
Kupffer cells and polymorphonuclear leukocytes (PMNs) contribute to the severe reperfusion injury of the liver after ischemia at different time points. The objective of this study was to identify the cellular source(s) of reactive oxygen formation during the PMN-induced injury phase. Kupffer cells and PMNs were isolated from the liver after 45 min of ischemia and 5 h or 24 h of reperfusion using collagenase-pronase digestion and a centrifugal elutriation method. Spontaneous superoxide anion (O2-) formation by large Kupffer cells (basal value 0.65 +/- 0.16 nmol/h/10(6) cells) was increased (up to 550%) during the entire reperfusion period. No enhanced O2- generation by the small Kupffer cell fraction was observed at any time. Control PMNs generated only small amounts of O2- spontaneously (0.25 +/- 0.05 nmol O2-/h/10(6) cells), but hepatic PMNs generated significantly more superoxide: 1.90 +/- 0.58 nmol O2-/h/10(6) cells at 5 h and similarly at 24 h of reperfusion. All cell types were significantly primed for enhanced O2- formation during reperfusion; the priming effect was consistently higher for stimulation with opsonized zymosan (receptor-mediated signal transduction pathway) compared to phorbol myristate acetate (protein kinase C activation). Our data support the hypothesis that PMNs and large Kupffer cells are predominantly responsible for the postischemic oxidant stress during the later reperfusion injury phase after hepatic ischemia in vivo.
Jaeschke
H
H
Department of Medicine, Baylor College of Medicine, Houston, TX 77030.
Bautista
A P
AP
Spolarics
Z
Z
Spitzer
J J
JJ
D000818:Animals; D007511:Ischemia / Q000378:metabolism*; D007728:Kupffer Cells / Q000187:drug effects / Q000378:metabolism*; D008099:Liver / Q000098:blood supply* / Q000473:pathology; D008297:Male; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D011916:Rats, Inbred F344; D017382:Reactive Oxygen Species / Q000378:metabolism; D015427:Reperfusion Injury / Q000378:metabolism*; D013268:Stimulation, Chemical; D013481:Superoxides / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D015054:Zymosan / Q000494:pharmacology
D017382:Reactive Oxygen Species; D013481:Superoxides; D015054:Zymosan; D013755:Tetradecanoylphorbol Acetate