Production of Prostaglandin F2α by Cultured Bovine Endometrial Cells in Response to Tumor Necrosis Factor α: Cell Type Specificity and Intracellular Mechanisms1

Abstract

Tumor necrosis factor alpha (TNFalpha) has been shown to be a potent stimulator of prostaglandin (PG) F(2alpha) secretion in the bovine endometrium. The aims of the present study were to determine the cell types in the endometrium (epithelial or stromal cells) responsible for the secretion of PGF(2alpha) in response to TNFalpha, and the intracellular mechanisms of TNFalpha action. Cultured bovine epithelial and stromal cells were exposed to TNFalpha (0.006-6 nM) or oxytocin (100 nM) for 4 h. TNFalpha resulted in a dose-dependent increase of PGF(2alpha) production in the stromal cells (P < 0.001) but not in the epithelial cells. On the other hand, oxytocin stimulated PGF(2alpha) output in the epithelial cells but not in the stromal cells. When the stromal cells were incubated for 24 h with TNFalpha and inhibitors of phospholipase (PL) C or PLA(2), only PLA(2) inhibitor completely stopped the actions of TNFalpha (P < 0.001). When the stromal cells were exposed to TNFalpha and arachidonic acid, the action of TNFalpha was augmented (P < 0.001). When the stromal cells were incubated for 24 h with a nitric oxide (NO) donor (S-NAP), S-NAP stimulated the PGF(2alpha) production dose-dependently. Although an NO synthase (NOS) inhibitor (L-NAME) reduced TNFalpha-stimulated PGF(2alpha) production, an inhibitor of phosphodiesterase augmented the actions of TNFalpha and S-NAP (P < 0. 05). The overall results indicate that the target of TNFalpha for stimulation of PGF(2alpha) production in cattle is the endometrial stromal cells, and that the actions of TNFalpha are mediated via the activation of PLA(2) and arachidonic acid conversion. Moreover, TNFalpha may exert a stimulatory effect on PGF(2alpha) production via the induction of NOS and the subsequent NO-cGMP formation.

The bovine endometrial epithelial cells promote the differentiation of trophoblast stem-like cells to binucleate trophoblast cells

“…However, EECs derived by trypsinization could not survive by passaging, and the stromal cell contamination was obtained by tissues cutting. Finally, we adopted the method described before (Horn et al 1998;Skarzynski et al 2000), but some modifications have been made. Compared with previously published methods (Horn et al 1998) of EEC derivation, our method simplified the process of digestion by reducing digestion time and adjusted the enzyme solutions from two (enzyme solution I: sterile 0.5 % pancreatin solution containing 4.8 mg/ml dispase II, enzyme solution II: 4.8 mg/ml dispase II in HBSS without Ca 2?…”

“…Primary bovine EECs were isolated and cultured as previously described (Horn et al 1998;Skarzynski et al 2000) with some modifications. Briefly, the healthy normal whole uteruses were removed from cows at a local abattoir within 30 min of death.…”

“…Therefore, the development of in vitro models is required to study implantation processes. Bovine endometrial epithelial cells (EECs) are often used as a bovine in vitro model for studying the biological characteristics, cellular and molecular mechanisms, and responses to infection or damage of the bovine uterine epithelium, as well as for investigating the impacts of bovine embryo implantation and development of interactions between the embryo and mother (Bai et al 2014;Fortier et al 1988;Hou et al 2015;Skarzynski et al 2000;Zhang et al 2013). Coculture of bovine EECs and bovine trophoblast cells can be used to investigate the maturation of trophoblasts and the formation of binucleate cells.…”

“…The isolation and culture of primary bovine EECs have been previously described in several reports (Horn et al 1998;Skarzynski et al 2000;Sakurai et al 2012). But the purity of endometrial cells has not been analyzed.…”

Inflammatory Mediators and Hormonal Changes in Subclinical and Clinically Affected Mastitis Cows

“…Severe cases of coliform mastitis are accompanied by the highest increase in blood plasma concentrations of both TNF-α and NO (Kahl et al, 1997;Hirvonen et al, 1999;Blum et al, 2000;Komine et al, 2004). Elevated TNF α in blood of mastitis animals Ohtuska et al, 2001) can increase PGF 2 α synthesis (Starzynski et al, 2000) and suppress LH surge leading to inhibition of fertilization and development of embryos (Hansen et al,2004).…”