Cellular mechanisms mediating the stimulation of ovine endometrial secretion of prostaglandin F2 alpha in response to oxytocin: role of phospholipase A2.

Four experiments were conducted to determine whether phospholipase (PL) A2 mediates the stimulatory effect of oxytocin on the release of prostaglandin (PG) F2 alpha from ovine endometrial tissue. Caruncular endometrial tissue was collected from ovariectomized ewes on the day after a steroid replacement protocol had been completed. The replacement protocol consisted of progesterone for 10 days (12 mg/day) followed by oestradiol on days 10 and 11 (100 micrograms/day) and had been shown previously to provide endometrial tissue that would release PGF2 alpha in response to oxytocin in vitro. In experiment 1, oxytocin (10(-7) M) and melittin (1.76 x 10(-6) M; a stimulator of PLA2) stimulated release of PGF2 alpha from tissue explants (P < 0.05). Aristolochic acid (10(-4) M; an inhibitor of PLA2) decreased oxytocin- and melittin-induced release of PGF2 alpha by 77% and 71% respectively (P < 0.05). Experiment 2 was conducted to establish the minimum inhibitory dose of aristolochic acid. Basal release of PGF2 alpha was inhibited at 10(-5) M aristolochic acid, but 10(-4) M was required to block the stimulatory effect of oxytocin. Experiment 3 was carried out to identify the precise intracellular locus at which aristolochic acid was exerting its effect. Oxytocin (10(-7) M), AlF4- (5 x 10(-2) M NaF, 10(-5) M AlCl3), melittin (1.76 x 10(-6) M) and arachidonic acid (AA; 20 micrograms/ml) stimulated release of PGF2 alpha (P < 0.05). Aristolochic acid (10(-4) M) blocked the release of PGF2 alpha stimulated by oxytocin, AlF4- or melittin by > 80% (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)