Elevated expression of p47phox and p67phox proteins in neutrophils from burned rats.

Abstract

The molecular control of neutrophil respiratory burst in burn injury was investigated through quantitation of protein factors, p47phox and p67phox, which are required for the activation of the phagocyte plasma membrane NADPH-oxidase. Circulating neutrophils were isolated from rats with 30% body surface area covered with full thickness burns. Neutrophil O2- generation, and p47phox and p67phox expressions, respectively, were determined using spectrophotometric and immunoblot techniques. Formylmethionyl-leucyl-phenylalanine stimulated superoxide anion generation was approximately 50% higher in neutrophils from rats 24 and 72 h after burns compared with that in sham control rats. The level of superoxide production was .47 +/- .05 nanomoles per minute per 10(6) cells (mean +/- SE, n = 6) at 24 h and .45 +/- .05 (n = 6) at 72 h postburn, whereas in sham control animals it was .32 +/- .02 (n = 8). Compared with the sham group p47phox levels, p47phox expression was 5.7-fold, 4.4-fold, and 4.5-fold higher, respectively, at 24, 36 and 72 h postburn. The levels of p67phox in burned animals were 2-fold higher than in the sham group, (p < .05) at 24 h postburn, and approximately 50% higher than sham at 36 h after the burn. The p67phox levels in rats 72 h after the burn were not significantly different from the sham values. These data support the occurrence of an up-regulation of p47phox and p67phox expressions accompanying the enhanced neutrophil respiratory burst activity during the early stages of burn injury. The up-regulation of p47phox and p67phox could be responsible for the priming of neutrophil O2- production leading to host tissue injury.