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Superoxide generation by neutrophils and Kupffer cells during in vivo reperfusion after hepatic ischemia in rats
Author(s) -
Jaeschke Hartmut,
Bautista Abraham P.,
Spolarics Zoltan,
Spitzer John J.
Publication year - 1992
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.52.4.377
Subject(s) - kupffer cell , superoxide , zymosan , biology , reperfusion injury , ischemia , pronase , endocrinology , medicine , microbiology and biotechnology , immunology , biochemistry , in vitro , enzyme , trypsin
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 die 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 (O 2 − ) 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 O 2 − generation by the small Kupffer cell fraction was observed at any time. Control PMNs generated only small amounts of O 2 − spontaneously (0.25 ± 0.05 nmol O 2 − /h/10 6 cells), but hepatic PMNs generated significantly more superoxide: 1.90 ± 0.58 nmol O 2 − /h/10 6 cells at 5 h and similarly at 24 h of reperfusion. All cell types were significantly primed for enhanced O 2 − formation during reperfusion; the priming effect was consistantly higher for stimulation with opsonized zymosan (receptor‐ mediated signal transduction pathway) compared to phor‐ bol 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 viva.