Reperfused gut elaborates PAF that chemoattracts and primes neutrophils.
Our in vivo model of mesenteric ischemia/reperfusion (I/R) has shown that the gut serves as a priming bed for neutrophils (PMN). Activation of phospholipase A2 (PLA2) during ischemia temporally precedes PMN sequestration in the gut and the appearance of primed PMN in the portal circulation. Therefore, we hypothesized that reperfused gut secretes platelet activating factor (PAF) via PLA2 activation that is responsible for increased PMN chemotaxis and priming for superoxide (O2-) generation. Sprague-Dawley rats underwent gut ischemia/reperfusion (45 min SMA occlusion/2 hr reperfusion) or sham laparotomy. Distal ileum was harvested, rinsed with bacteriostatic saline/neomycin, and incubated for 1 hr at 37 degrees C in RPMI 1640 and the cell-free supernatant was collected. Normal human PMNs, isolated by plasma-Percoll gradients, were pretreated with or without a PAF receptor antagonist (WEB 2170). Chemotaxis toward gut supernatant was then measured by the agarose method. Additionally, PMNs were preincubated with or without WEB 2170 and their O2- release in response to 1 microM FMLP was measured by the Vmax of SOD-inhibitable cytochrome c reduction. Reperfused gut produced a chemotactic index of 2.1 +/- 0.1 compared to 0.2 +/- 0.9 following sham laparotomy (P < 0.05); this was reduced to 0.4 +/- 0.9 with PAF receptor blockade. Similarly, gut I/R supernatant primed PMNs for O2- (P < 0.05) compared to laparotomy, and this effect was abrogated by a PAF antagonist. These data suggest that reperfused gut can elaborate PAF which chemoattracts and primes PMNs for O2- generation.
Kim
F J
FJ
Department of Surgery, Denver General Hospital, Colorado 80204, USA.
Moore
E E
EE
Moore
F A
FA
Biffl
W L
WL
Fontes
B
B
Banerjee
A
A
D000818:Animals; D002634:Chemotaxis, Leukocyte; D007422:Intestines / Q000098:blood supply*; D008297:Male; D009504:Neutrophils / Q000276:immunology / Q000502:physiology*; D010741:Phospholipases A / Q000502:physiology; D054467:Phospholipases A2; D010972:Platelet Activating Factor / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015424:Reperfusion; D013481:Superoxides / Q000378:metabolism
D010972:Platelet Activating Factor; D013481:Superoxides; D010741:Phospholipases A; D054467:Phospholipases A2