The aim of this in vitro study was to examine the release of progesterone by porcine ovarian granulosa cells (GCs) after exposure to toxic concentrations of deoxynivalenol (DON), resveratrol (RSV), and their combination (DON with RSV). Ovarian granulosa cells were incubated without (control) or with treatments of natural substances at various doses for 24 h: RSV (10, 30 and 50 μg/mL) / DON (2000, 3000 and 5000 ng/mL), and their combination (10 μg/mL of RSV with 2000 ng/mL of DON; 30 μg/mL of RSV with 3000 ng/mL of DON; 50 μg/mL of RSV with 5000 ng/mL of DON). Progesterone was determined by radioimmunoassay (RIA). Progesterone release was significantly (P < 0.05) stimulated by RSV at the doses 50 μg/mL but not at 30 and 10 μg/mL and by DON treatment at all used doses (2000, 3000 and 5000 ng/mL). RSV in combination with DON stimulated significantly (P < 0.05) the progesterone release by GCs at the highest doses (50 μg/mL of RSV with 5000 ng/mL of DON). On the other hand, the stimulatory effect of RSV in combination with DON was significantly (P < 0.05) lower in comparison with alone DON effect. In conclusion, our results indicate, (1) the dose-depended stimulatory effects of RSV, DON and combination of RSV with DON on release of steroid hormone progesterone and (2) reduction of the stimulatory effect of DON by RSV. Our in vitro results suggest that reproductive toxicity of animals induced by a mycotoxin - deoxynivalenol can be inhibited by a protective natural substance - resveratrol.
The aim of this study was to examine the effect of A-trichothecenes T-2 and HT-2 toxins combined with insulin-like growth factor I (IGF-I) on the release of steroid hormone progesterone (P4) by porcine ovarian granulosa cells (GCs). The cells were incubated without (control) or with treatments of A-trichothecenes T-2 (100 and 1000 ng/mL)/ HT-2 (100 and 1000 ng/mL) combined with IGF-I (1, 10 and 100 ng/mL) for 24 h. Progesterone secretion was determined by RIA. The release of P4 by GCs after addition of T-2 toxin (at 100 ng/mL) combined with IGF-I (at 10 but not at 1 and 100 ng/mL) and HT-2 toxin (at 100 ng/mL) combined with IGF-I (at all doses) was significantly (P < 0.05) inhibited. On the other hand the release of P4 after addition of T-2/ HT-2 toxin (at 1000 ng/mL) combined with IGF-I (at all doses) was significantly (P < 0.05) stimulated. Alone IGF-I addition (at 10, 100 but not at 1 ng/mL) significantly (P < 0.05) stimulated P4 release by GCs. The results of our in vitro study indicate the T-2 and HT-2 toxins combined with IGF-I could modify progesterone secretion by porcine ovarian granulosa cells and potentially regulate process of steroidogenesis in the ovaries. Currently, occurrence of mycotoxins in food and feed is a worldwide problem and therefore study of these toxins as well as their interaction with different substances such as growth factors could be beneficial for better understanding of mechanism of their toxic effects in organism.
The possible effects of a natural substance amygdalin and its combination with the mycotoxin deoxynivalenol (DON) on the steroid hormone secretion (progesterone and 17-β-estradiol) by porcine ovarian granulosa cells (GCs) were examined in this in vitro study. Ovarian GCs were incubated without (control group) and with amygdalin (1, 10, 100, 1,000 and 10,000 μg mL(1)), or its combination with DON (1 μg mL(1)) for 24 h. The release of steroid hormones was determined by ELISA. The progesterone secretion by porcine ovarian GCs was not affected by amygdalin in comparison to the control. However, the highest amygdalin dose (10,000 μg mL(1)) caused a significant stimulation of the 17-β-estradiol release. A combination of amygdalin with DON significantly (P < 0.05) increased the progesterone release at all concentrations. Similarly, a stimulatory effect of amygdalin co-administered with DON was detected with respect to the 17-β-estradiol secretion at the highest dose (10,000 μg mL(1)) of amygdalin and 1 μg mL(1) of DON. Noticeable differences between the effects of amygdalin alone and its combination with DON on the progesterone release were detected. In contrast, no differences between the stimulatory effects of amygdalin and its combination with DON on the 17-β-estradiol synthesis by porcine GCs were observed. Findings from this in vitro study did not confirm the expected protective effect of amygdalin on mycotoxin induced reprotoxicity. Our results indicate that the stimulatory effect of amygdalin combined with DON on the progesterone release was clearly caused by the DON addition, not by the presence amygdalin per se. On the other hand, the stimulation of 17-β-estradiol production was solely caused by the presence of amygdalin addition. These findings suggest a possible involvement of both natural substances into the processes of steroidogenesis and appear to be endocrine modulators of porcine ovaries.
This report provides information about the impact of chosen natural plant extracts on basic ovarian functions. This article summarizes our results concerning the effect of selected plant extracts on proliferation, apoptosis and hormone secretion – release of progesterone (P4), testosterone (T) and leptin (L) on porcine granulosa cells (GC), We analyzed effects of ginkgo (GB), rooibos (RB), flaxseed (FL), green tea polyphenols (GTPP), green tea - epigallocatechin-3-gallate (EGCG), resveratrol (RSV) and curcumin (CURC) (0; 1; 10 and 100 μg.ml-1) on markers of proliferation, apoptosis and secretory activity of porcine ovarian granulosa cells by using immunocytochemistry and EIA. It was demonstrated, that all these natural plants and plant molecules inhibited the accumulation of proliferation-related peptide (PCNA) and apoptosis-associated peptide (Bax) in cultured. Furthermore, it was observed that natural plant extracts altered progesterone, testosterone and leptin release in porcine ovarian cells. It is concluded, that GB, RB, FL, RSV, CURC, GTPP and EGCG can directly affect ovarian cells and therefore they could potentially influence ovarian functions.
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