Neutrophil lipoxygenase activation and leukosequestration in postischemic myocutaneous flaps: role of ltb4. Kirschner, Richard E., Jerry J. C. Chiao, Billie S. Fyfe, Lloyd A. Hoffman, John Mihran Davis, and Gary A. Fantini. Department of Surgery, The New York Hospital and Cornell University Medical College, New York, New York 10021; and Department of Pathology, The Mount Sinai School of Medicine, New York, New York 10029
APStracts 2:0057H, 1995.
Reperfusion of ischemic tissues leads to eicosanoid- and polymorphonuclear leukocyte (PMN)-dependent injury. The present experiments were undertaken to examine the effect of myocutaneous flap ischemia-reperfusion on neutrophil 5-lipoxygenase (5-LO) activity and to define the role of leukotriene B4 (LTB4) in postischemic PMN infiltration into such composite tissue grafts. Anesthetized Yorkshire pigs underwent six hours of rectus abdominis myocutaneous flap ischemia or sham ischemia, and LTB4 generation was measured in calcium ionophore-stimulated neutrophils isolated from the circulation. At 30 minutes of reperfusion, neutrophil generation of LTB4 increased from a baseline value of 31.0 + 6.8 ng/5 X 106 PMN to 98.5 + 5.1 ng/5 X 106 PMN (P < 0.01) and was significantly greater than that of neutrophils isolated from animals subjected to sham ischemia and reperfusion (54.3 + 4.1 ng/5 X 106 PMN; P < 0.01). Pretreatment of animals with the LTB4 receptor antagonist SC -41930 (n = 5) significantly attenuated reperfusion-associated 5-LO activation (60.3 + 11.6 ng LTB4 /5 X 106 PMN; P < 0.01), suggesting the presence of a positive feedback mechanism for eicosanoid biosynthesis. Graft ischemia in control animals was associated with progressive PMN infiltration at one and four hours of reperfusion (334 + 92 and 667.2 + 198 PMNs/25 HPF, respectively); however, pretreatment with SC-41930 or the 5-LO inhibitor diethylcarbamazine dramatically reduced PMN infiltration at one hour of reperfusion (22 + 3 and 64 + 36 PMNs/25 HPF, respectively; P < 0.01) and at four hours of reperfusion (178 + 31 and 156 + 40 PMNs/25 HPF; P < 0.01). These data indicate that myocutaneous flap ischemia-reperfusion leads to activation of 5-LO in circulating PMNs and suggest that LTB4 plays a critical role in the pathogenesis of reperfusion injury by amplifying the process of neutrophil recruitment into postischemic tissues.

Received 5 April 1994; accepted in final form 28 November 1994.
APS Manuscript Number H310-4.
Article publication pending Am. J. Physiol. (Heart Circ. Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on  7 March 1995.