Abstract
Fatty acids have been involved in the proliferation and differentiation of numerous cells, as mediated via peroxisome proliferator-activated receptors (PPARs) or lipid metabolites (prostaglandins, diacylglycerol). In the present study, we have investigated the effect of arachidonic acid (AA), docosahexaenoic acid (DHA) and its precursor eicosapentaenoic acid (EPA) on the differentiation of a rat uterine stromal cell line, UIII. As markers of decidualization, we have investigated morphological changes, monitored by inverted light and scanning electron microscopy. The induction of 3 proteins, desmin, hsp-25 and prolactin, which are all considered to be markers of decidualization, were analyzed by immunocytochemistry or Western blotting. Addition of AA (30 microM) to the medium of cultured cells for 48h induced cell spreading and flattening. Cells became enlarged (x 2.5) and some of them were binucleated. Using scanning electron microscopy, we confirmed these morphological changes and showed that the enlargement of the cells was followed by numerous extracellular processes, leading to an increase in cell surface area and intercellular communications. Immunocytochemistry showed that this treatment also induced the expression of desmin, which seems to direct morphological changes, beginning as a perinuclear ring and extending to the cell membrane. The time course of desmin expression was studied by Western blotting. No desmin expression was present before 4h of AA treatment. Desmin induction was maximum at 24h of treatment and plateaued thereafter. DHA and EPA (30 microM), added to the medium, failed to induce any change. However, in cells previously differentiated with AA and expressing desmin, treatment with DHA or EPA (30microM) reversed partially the action of AA, EPA being the most effective. AA also induced hsp-25, though all cells did not express this protein. A prolactin (PRL)-like factor was induced by AA, as recognized by an antibody against pituitary rPRL, and migrated as the standard. Moreover, a fragment of 16 kDa was also revealed by this antibody, suggesting that the PRL-like factor cleaved, was similar to PRL and that the PRL-like factor could be identical to PRL. In conclusion, these results show that AA is able to specifically induce the decidualization of uterine stromal cells in vitro.
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