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in Bovine Endometrial Cells1
a Centre de recherche en reproduction animale, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Quebec, Canada J2S 7C6
Oxytocin (OT) is responsible for the episodic release of luteolytic prostaglandin (PG) F2
from the uterus in ruminants. The attenuation of OT-stimulated uterine PGF2 secretion by interferon-
(IFN-) is essential for prevention of luteolysis during pregnancy in cows. To better understand the mechanisms involved, the effect of recombinant bovine IFN- (rbIFN-) on OT-induced PG production and cyclooxygenase-2 (COX-2) and PGF synthase (PGFS) expression in cultured endometrial epithelial cells was investigated. Cells were obtained from cows at Days 1–3 of the estrous cycle and cultured to confluence in RPMI medium supplemented with 5% steroid-free fetal calf serum. The cells were then incubated in the presence or absence of either 100 ng/ml OT or OT+100 ng/ml rbIFN- for 3, 6, 12, and 24 h. OT significantly increased PGF2 and PGE2 secretion at all time points (p < 0.01), while rbIFN- inhibited the OT-induced PG production and reduced OT receptor binding in a time-dependent manner. OT increased the steady-state level of COX-2 mRNA, measured by Northern blot, which was maximal at 3 h (9-fold increase) and then decreased with time (p < 0.01). OT also caused an increase in COX-2 protein, which peaked at 12 h (11-fold increase), as measured by Western blot. Addition of rbIFN- suppressed the induction of COX-2 mRNA (89%, p < 0.01) and COX-2 protein (50%, p < 0.01) by OT. OT also increased PGFS mRNA, and this stimulation was attenuated by rbIFN- (p < 0.01). To ensure that the decrease in COX-2 was not solely due to down-regulation of the OT receptor, cells were stimulated with a phorbol ester (phorbol 12-myristate 13-acetate; PMA) in the presence and absence of rbIFN-. The results showed that rbIFN- also decreased PMA-stimulated PG production and COX-2 protein. It can be concluded that rbIFN- inhibition of OT-stimulated PG production is due to down-regulation of OT receptor, COX-2, and PGFS.
2 Correspondence: Alan K. Goff, Centre de recherche en reproduction animale, Faculté de médecine vétérinaire, Université de Montréal, 3200 rue Sicotte, St-Hyacinthe, PQ, Canada J2S 7C6. FAX: 450 778 8103; goffak{at}medvet.umontreal.ca
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