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Interferon- Induces Degradation of Prostaglandin H Synthase-2 Messenger RNA in Bovine Endometrial Cells Through a Transcription-Dependent Mechanism¹

Department of Animal Sciences³ Department of Microbiology and Cell Science,⁴ University of Florida, Gainesville, Florida 32611-0910

A series of experiments were undertaken to examine the effects of interferon (IFN)- on regulation of prostaglandin H synthase (PGHS)-2 mRNA in bovine endometrial (BEND) cells as a means to elucidate the actions of IFN- to maintain pregnancy. The objective was to determine if IFN- mediates posttranscriptional regulation of PGHS-2 mRNA. Cells were treated with phorbol 12,13-dibutyrate (PdBu) for 3 h to induce PGHS-2 mRNA expression. Actinomycin D (0 or 1 µg/ml) or the p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580 (1 µM), were added at 3 h, followed by addition of IFN- (0 or 50 ng/ml) at 3.5 h and extraction of RNA at 4.5 h. The concentrations of PGHS-2 mRNA were stable between 3 and 4.5 h regardless of actinomycin D. Simultaneous treatment of PdBu-treated cells with actinomycin D and SB203580 (1 µM) decreased PGHS-2 mRNA. Addition of IFN- (50 ng/ml) reduced PGHS-2 mRNA, which was not observed when actinomycin D was present. Concurrent treatments of cells with SB203580 and IFN- (5 ng/ml) decreased concentrations of PGHS-2 mRNA in an additive manner. Although IFN- reduced PGHS-2 mRNA concentrations, phosphorylation of p38 MAPK was induced by IFN-, PdBu, and PdBu combined with IFN- after 10 min of treatment. Both the p38 MAPK inhibitor and IFN- decreased prostaglandin F₂ secretion, and decreases were additive when the two were given together. In summary, activation of p38 MAPK by PdBu is required for continued presence of PGHS-2 mRNA and secretion of prostaglandin F₂ in BEND cells. Interferon- mediates a transcription-dependent mechanism, which induces degradation of PGHS-2 mRNA. However, the consequences of an IFN--induced activation of p38 MAPK warrant further investigation, because inhibition of p38 MAPK caused a degradation of PGHS-2 mRNA.

¹ Supported in part by the NRI Competitive Grant Program/USDA, grant 98-35203-6367. This is Florida Agricultural Experimental Station Journal Series R-10035.

² Correspondence: William W. Thatcher, Department of Animal Sciences, P.O. Box 110920, Gainesville, FL 32611-0910. FAX: 352 392 5595; thatcher{at}animal.ufl.edu

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