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The aim of the study was to establish the effect of the ovarian steroids: 17β-oestradiol (E2) and testosterone (T4) on OT-stimulated PGF2α and PGE2 secretion by bovine endometrial cells. The epithelial endometrial cells from days 14-18 of the oestrous cycle (10⁵/ml) were incubated in DMEM/Ham's F12 with 10% FCS (38°C, atmosphere of air and 5% CO₂) for 72-96 h and the last 24 h in DMEM/Ham's F12 with 0.1% BSA. In Exp.l, the doses of steroids used to study their effect on the secretion of PGF2α and PGE2 from endometrial cells stimulated by OT were determined. In Exp.2, the cells were pre-incubated for 30 min with selected doses of steroids: P4 (10⁻⁵ M), T4 (10⁻⁵ M), and E2 (10⁻⁸ M) and next for 4 d with: arachidonic acid (AA; 10⁻⁵ M), OT (10⁻⁷ M) and OT with each of these steroids. The concentration of PGE2 and PGFM -metabolite of PGF2α was determined by EIA. P4, T4, and E2 did not affect (P>0.05) the basal secretion of PGF2α and PGE2, but all the steroids inhibited (P<0.01) OT- stimulated PGF2α secretion. The stimulating effect of OT on PGE2 secretion was not affected by P4 and T4 (P>0.05). This data suggests that different cellular mechanisms exist for steroids affecting the secretion of both prostaglandins from endometrial cells. Moreover, we suggest that non-genomic effect of P4 on bovine endometrial cells is non-specific since the other steroids can impair the effect of OT on these cells. This effect of the steroids can directly modulate function of endometrial cells.
The goal of this study was to evaluate the effect of various concentrations of interferon-tau (IFN-τ) with or without steroid hormones, 17ß estradiol or progesterone, on the proliferation of bovine endometrial cells in vitro. Endometrial epithelial and stromal cells were isolated from the uterus of cows during the early estrus cycle (2-3 days) and incubated with different doses of IFN-τ with or without steroid hormones. The proliferation was determined by the MTT test in 48, 96, and 144 h of incubation. An antiproliferative activity of IFN-τ was observed both in epithelial and stromal cells cultured in RPMI 1640 medium supplemented with 10% FBS or serum replacement. However, epithelial cells were more sensitive to antiproliferative action of interferon-tau. It;s activity was dose- and time-dependent. The inhibition of epithelial cell proliferation by 50% (ED50) was achieved at concentrations of 500 U/ml, 340 U/ml, and 8.8 U/ml of IFN-τ after 48, 96, and 144 h of incubation, respectively. None of the doses of IFN-τ (10-10.000 U/ml) used inhibited stromal cell proliferation in 50%. The most effective dose of IFN-τ inhibiting stromal cell proliferation was 10.000 U/ml, which decreased cell growth by 17.08%, 22.87%, and 2.6% after 48, 96, and 144 h of incubation, respectively. Steroid hormones, 17ß estradiol and progesterone, added to the culture of stromal cells with or without IFN-τ did not significantly modulate stromal cell growth. In contrast, a high concentration of progesterone (10-5M) alone significantly enhanced stromal cell growth. Progesterone at low, physiological concentrations (107 - 10-9 M) ameliorated the antiproliferative activity of IFN-τ, especially at the 109 M concentration. At this concentration, the stimulatory effect on stromal cell growth was observed. The mechanisms of such response are not entirely clear but may arise from the influence of IFN-τ on progesterone down regulation of its own receptor. Depicted activity of IFN-τ may find usefulness in therapy of neoplastic disorders.
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