Hyperbaric oxygen decreases ischemia-reperfusion–induced neutrophil/intercellular adhesion molecule-1 adhesion by blocking CD18 polarization. The purpose of this study was to evaluate whether this hyperbaric oxygen effect is nitric oxide dependent and to determine whether nitric oxide synthase is required.
A gracilis muscle flap was raised in nine groups of male Wistar rats. Global ischemic injury was induced by clamping the gracilis muscle pedicle artery and vein for 4 hours. The hyperbaric oxygen treatment consisted of 100% oxygen at 2.5 atm absolute during the last 90 minutes of ischemia. Groups were repeated with and without various nitric oxide synthase inhibitors and carboxy-2-phenyl-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide (C-PTIO), a nitric oxide scavenger. Normal neutrophils were exposed to activated plasma on intercellular adhesion molecule-1–coated coverslips (percentage adherent) and labeled with fluorescein isothiocyanate/antirat-CD11b for confocal microscopy (percentage polarized). The percentage of adherent and polarized cells was reported as mean ± SEM. Statistical analysis was by analysis of variance. A value of p ≤ 0.05 was considered significant.
C-PTIO–treated ischemia-reperfusion/hyperbaric oxygen plasma showed a significant increase in the percentage polarization of CD18 compared with ischemia-reperfusion/hyperbaric oxygen–untreated plasma from 4.1 ± 2.5 percent to 33.7 ± 7.7 percent (p ≤ 0.05). The nitric oxide scavenger C-PTIO also increased the percentage of adherent cells from 1.6 ± 0.4 percent to 20.3 ± 5.9 percent (p ≤ 0.05). Administration of NG-nitro-L-arginine methyl ester and other nitric oxide synthase inhibitors before hyperbaric oxygen treatment restored neutrophil adhesion and CD18 polarization to ischemia-reperfusion control values, significantly greater than ischemia-reperfusion/hyperbaric oxygen alone.
These results suggest that the hyperbaric oxygen reduction of ischemia-reperfusion–induced neutrophil polarization of CD18 and adherence to intercellular adhesion molecule-1 is mediated through a nitric oxide mechanism that requires nitric oxide synthase.