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by Cultured Bovine Endometrial Cells in Response to Tumor Necrosis Factor : Cell Type Specificity and Intracellular Mechanisms1
a Laboratory of Reproductive Endocrinology, Faculty of Agriculture, Okayama University, Okayama 700–8530, Japan
Tumor necrosis factor 
(TNF) has been shown to be a potent stimulator of prostaglandin (PG) F2 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 PGF2 in response to TNF, and the intracellular mechanisms of TNF action. Cultured bovine epithelial and stromal cells were exposed to TNF (0.006–6 nM) or oxytocin (100 nM) for 4 h. TNF resulted in a dose-dependent increase of PGF2 production in the stromal cells (P < 0.001) but not in the epithelial cells. On the other hand, oxytocin stimulated PGF2 output in the epithelial cells but not in the stromal cells. When the stromal cells were incubated for 24 h with TNF and inhibitors of phospholipase (PL) C or PLA2, only PLA2 inhibitor completely stopped the actions of TNF (P < 0.001). When the stromal cells were exposed to TNF and arachidonic acid, the action of TNF 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 PGF2 production dose-dependently. Although an NO synthase (NOS) inhibitor (L-NAME) reduced TNF-stimulated PGF2 production, an inhibitor of phosphodiesterase augmented the actions of TNF and S-NAP (P < 0.05). The overall results indicate that the target of TNF for stimulation of PGF2 production in cattle is the endometrial stromal cells, and that the actions of TNF are mediated via the activation of PLA2 and arachidonic acid conversion. Moreover, TNF may exert a stimulatory effect on PGF2 production via the induction of NOS and the subsequent NO-cGMP formation.
1 This research was supported by Grants-in-Aid for Scientific Research (No. 11556054 and 11460129) from the Ministry of Education, Science, Sports and Culture of Japan. D.J.S. was a postdoctoral fellow supported by the Japan Society for the Promotion of Science (JSPS). Y.M. is a research fellow of JSPS (No. 07809).
2 Correspondence. FAX: 81 86 251 8388; kokuda{at}cc.okayama-u.ac.jp
3 Current address: Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10–718 Olsztyn-Kortowo, Poland.
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