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Differential Effects of Interferon- on the Prostaglandin Synthetic Pathway in Bovine Endometrial Cells Treated with Phorbol Ester^*

Department of Animal Sciences, University of Florida, Gainesville 32611

Corresponding author: W. W. Thatcher; e-mail: thatcher{at}animal.ufl.edu.

Rationale for these experiments was to evaluate the dose effects of bovine interferon- (IFN-) on the prostaglandin secretory pathway of immortalized bovine endometrial (BEND) cells in response to phorbol 12, 13-dibutyrate (PdBu) and to characterize similar responses in primary bovine uterine epithelial cells as a biomonitor of embryo-induced antiluteolytic effects on the endometrium. The BEND cells were treated with PdBu (0 or 100 ng/mL) and IFN- (0 or 50 ng/mL) for 6 h. The PdBu stimulated secretions of PGF₂ and prostaglandin E₂ (PGE₂). Co-treatment of cells with IFN- blocked PdBu-induced secretion of both PGF₂ and PGE₂. Treatment with PdBu for 6 h induced expression of prostaglandin H synthase-2 mRNA, prostaglandin H synthase-2 protein, and prostaglandin E synthase mRNA, which were blocked with concurrent addition of IFN-. Doses of IFN- (0.05, 0.5, 1, 5, 10, and 20 µg/mL) were used with PdBu (0 and 100 ng/mL). The IFN- alone failed to stimulate secretion of PGF₂ and PGE₂, whereas IFN- doses <5 µg/mL suppressed PdBu-stimulated secretions of PGF₂ and PGE₂. Uterine epithelial cells were isolated from cows at d 17 after estrus and were cultured to confluence in serum-free medium. Cells were treated with IFN- (0, 50, or 500 ng/mL) and PdBu (0 or 100 ng/mL) before media were collected after 24 h for PGF₂ and PGE₂ analyses. Treatment of primary uterine epithelial cells with PdBu induced PGF₂ secretion, and IFN- (50 and 500 ng/mL) caused a reduction in PGF₂ secretion induced by PdBu. In the absence of PdBu, IFN- increased basal secretion of PGF₂. Concentrations of PGE₂ increased in response to PdBu, and the 50-ng/mL dose of IFN- had a stimulatory effect on PGE₂ concentrations compared with the 500-ng/mL dose in the absence of PdBu. Phorbol ester-induced gene transcription as related to prostaglandin synthesis is regulated by IFN- in vitro.

Key Words: bovine • endometrial cells • interferon- • prostaglandins

Abbreviation key: BEND = immortalized bovine endometrial, GAPDH = glyceraldehyde 3-phosphate dehydrogenase, IFN- = interferon-, IRF-1 = interferon regulatory factor-1, PdBu = phorbol 12, 13-dibutyrate, PGE₂ = prostaglandin E₂, PGES = prostaglandin E synthase, PGHS-2 = prostaglandin H synthase-2, PKC = protein kinase C

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