Neutrophilic cell-free exudate induces antinociception mediate by the protein S100A9.

Calcium-binding protein S100A9 (MRP-14) induces antinociceptive effect in an experimental model of painful sensibility and participates of antinociception observed during neutrophilic peritonitis induced by glycogen or carrageenan in mice. In this study, the direct antinociceptive role of the protein S100A9 in neutrophilic cell-free exudates obtained of mice injected with glycogen was investigated. Mice were intraperitoneally injected with a glycogen solution, and after 4, 8, 24, and 48 hours, either the pattern of cell migration of the peritoneal exudate or the nociceptive response of animals was evaluated. The glycogen-induced neutrophilic peritonitis evoked antinociception 4 and 8 hours after inoculation of the irritant. Peritoneal cell-free exudates, collected in different times after the irritant injection, were transferred to naive animals which were submitted to the nociceptive test. The transference of exudates also induced antinociceptive effect, and neutralization of S100A9 activity by anti-S100A9 monoclonal antibody totally reverted this response. This effect was not observed when experiments were made 24 or 48 hours after glycogen injection. These results clearly indicate that S100A9 is secreted during glycogen-induced neutrophilic peritonitis, and that this protein is responsible by antinociception observed in the initial phase of inflammatory reaction. Thus, these data reinforce the hypothesis that the calcium-binding protein S100A9 participates of the endogenous control of inflammatory pain.

Evidence that glutamine is involved in neutrophil function.

Phosphate-dependent glutaminase (PDG) activity, a key enzyme of glutamine metabolism, was determined in neutrophils obtained from the intra-peritoneal cavity (PC) or bronchoalveolar space (BAS) after administration of 1 ml or 100 microl, respectively of saline, glycogen solution (1%) or lipopolysaccharide (LPS 0.1 mg (100 microl)(-1)). Neutrophils were obtained by lavage of both sites with 20 ml saline 24 h after the administration of the stimuli. Glycogen and LPS, depending on the site the cells were obtained from, differently modulated PDG activity. Cells from BAS stimulated by glycogen or LPS had raised PDG activity to 30.5 +/- 5.2 and 42.7 +/- 12.1 nmol min(-1) mg(-1) protein, respectively, when compared with saline (9.1 +/- 0.9 nmol min(-1) mg(-1) protein); mean +/- SEM. On the other hand, cells from PC showed different PDG activity: 52.0 +/- 12.6 nmol min(-1) mg(-1) for saline, 36.5 +/- 9.5 nmol min(-1) mg(-1) for glycogen, and 76.6 +/- 11.2 nmol min(-1) mg(-1) for LPS; mean +/- SEM. Therefore, PDG activity varies with the site from which neutrophils are obtained and the stimulus imposed. The effect of glutamine on nitric oxide (NO) and tumour necrosis factor (TNF) production by peritoneal neutrophils, obtained after glycogen administration, cultured in the presence of LPS (0.5 microg ml(-1)) was also examined. The addition of glutamine at concentrations varying from 2 to 20 mM did not markedly affect NO production. Glutamine alone at 2 mM did not modify the production of TNF but in the presence of LPS caused a significant decrease. So, glutamine may preserve the function of neutrophils during infections and injuries.

Endometritis, salpingitis and fertilisation rates after mating mares with a history of intrauterine lumenal fluid accumulation.

The occurrence of uterine and oviductal inflammation, and fertilisation rates, were measured on Day 3 post ovulation in inseminated mares that had either exhibited intrauterine lumenal fluid during a previous dioestrus (Experiment 1) or had acute endometritis induced by intrauterine infusion of 1% glycogen (Experiment 2). Endometritis was assessed by uterine cytology and histology whereas oviductal inflammation was measured histologically. Fertilisation rates were calculated from the percentage of cleaved ova recovered by retrograde flushing of the oviducts. Mares with or without pre-existing uterine fluid during dioestrus that were inseminated showed a higher incidence of endometritis than control mares without pre-existing uterine fluid that were not inseminated (n = 7 mares/group). However, inseminated mares with uterine fluid did not show a higher incidence of endometritis than inseminated mares without uterine fluid. Mares with or without pre-existing uterine fluid showed a higher incidence of endometritis than salpingitis and these 2 groups of mares showed equivalent rates of fertilisation and oviductal oocyte recovery. Mares inseminated with semen alone or semen following 1% glycogen treatment had a higher incidence of endometritis than control noninseminated mares (n = 17 mares/group) but mares that received semen plus 1% glycogen did not show a higher incidence of endometritis than mares that received semen alone. Both these groups of mares showed a higher incidence of endometritis than salpingitis and those that received semen plus 1% glycogen showed an equal recovery rate of recently ovulated ova but a lower fertilisation rate than the mares that received semen alone.