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Stochmal'ova A.,Constantine the Philosopher University | Kadasi A.,Slovak University of Agriculture | Alexa R.,Constantine the Philosopher University | Grossmann R.,Institute of Animal Science | And 2 more authors.
Potravinarstvo | Year: 2014

Yucca shidigera is a medicinal plant native to Mexico. Is a plant widely used in folk medicine to treat a variety of ailmentary disorders, but its action on reproductive processes and possible mechanisms of such action remains unknown. Yucca schidigera extract contains a number of steroidal saponins that, because of their biological activity, have attracted attention from the food industry for many years. Yucca extract is used as a natural feed additive with positive effect to microflora, digestion, metabolism and to improve animal muscle growth. Its extract has been used as a foodstuff and folk medicine to treat a wide variety of diseases for many years. Nevertheless, it remaines unknown, whether consumption of yucca can affect reproductive system. The aim of this study was to examine the effects of yucca on basic ovarian cell functions-proliferation, apoptosis and steroidogenesis. Porcine ovarian granulosa cells were cultured with and without yucca extract (added at doses 0; 1; 10 and 100 μg.mL-1 of medium). Markers of proliferation (% of PCNA-positive cells) and apoptosis (% cells containing bax) were analysed by immunocytochemistry. Release of steroid hormones (progesterone and testosterone) was measured by EIA. It was observed, that addition of yucca inhibited proliferation (expression of PCNA), increased apoptosis (expression of bax), stimulated progesterone and inhibited testosterone release. The ability of yucca to reduce ovarian cell proliferation, to promote ovarian cell apoptosis and affect steroidogenesis demonstrates the direct influence of yucca on female gonads. Furthermore, our observations suggest the multiple sites of action (proliferation, apoptosis, steroidogenesis) of yucca on porcine ovarian cell functions. It is not to be excluded, that consumption of yucca can suppress female reproductive functions. © 2014 Potravinarstvo. Source


Kolesarova A.,Slovak University of Agriculture | Capcarova M.,Slovak University of Agriculture | Medvedova M.,Slovak University of Agriculture | Sirotkin A.V.,Institute for Genetics and Reproduction of Farm Animals | Kovacik J.,Slovak University of Agriculture
Physiological Research | Year: 2011

It would be desirable to expand the existing general knowledge concerning direct action of metals on the ovary. Nevertheless, the results of testing of iron compound on porcine ovarian cells should be interpreted carefully because iron is an essential element which could also induce changes in cellular processes. The aim of this in vitro study was 1) to examine dose-dependent effects of iron on the secretory activity of porcine ovarian granulosa cells, and 2) to outline the potential intracellular mediators mediating these effects. Specifically, we evaluated the effect of iron sulphate on the release of insulin-like growth factor I (IGF-I) and progesterone, as well as the expression of markers of proliferation (cyclin B1) and apoptosis (caspase-3) in porcine ovarian granulosa cells. Concentrations of IGF-I and progesterone were determined by RIA, cyclin B1 and caspase-3 expression by immunocytochemistry (ICC). Our results show a significantly decreased IGF-I secretion by ovarian granulosa cells after iron sulphate addition at the doses 0.5 and 1.0 mg/ml. The iron sulphate additions at doses 0.17 and 1.0 mg/ml had no effect on progesterone secretion. In contrast, iron sulphate addition at doses 0.17-1.0 mg/ml resulted in stimulation of cyclin B1 and caspase-3 expression. In conclusion, the present results indicate a direct effect of iron on 1) secretion of growth factor IGF-I but not steroid hormone progesterone, 2) expression of markers of proliferation (cyclin B1), or 3) apoptosis (caspase-3) of porcine ovarian granulosa cells. These results support an idea that iron could play a regulatory role in porcine ovarian function: hormone release, proliferation and apoptosis. © 2011 Institute of Physiology v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic. Source


Kolesarova A.,Slovak University of Agriculture | Bakova Z.,Slovak University of Agriculture | Capcarova M.,Slovak University of Agriculture | Galik B.,Slovak University of Agriculture | And 4 more authors.
Journal of Animal Physiology and Animal Nutrition | Year: 2013

The aim of this study was to examine possible effects of bee pollen added to the feed mixture (FM) on rat ovarian functions (secretion activity and apoptosis). We evaluated the bee pollen effect on the release of insulin-like growth factor I (IGF-I) and steroid hormones (progesterone and estradiol), as well as on the expression of markers of apoptosis (Bcl-2, Bax and caspase-3) in rat ovarian fragments. Female rats (n=15) were fed during 90days by FM without or with rape seed bee pollen in dose either 3kg/1000kg FM or 5kg/1000kg FM. Fragments of ovaries isolated from rats of each group (totally 72 pieces) were incubated for 24h. Hormonal secretion into the culture medium was detected by RIA. The markers of apoptosis were evaluated by Western blotting. It was observed that IGF-I release by rat ovarian fragments was significantly (p<0.05) decreased; on the other hand, progesterone and estradiol secretion was increased after bee pollen treatment at dose 5kg/1000kg FM but not at 3kg/1000 FM. Accumulation of Bcl-2 was increased by bee pollen added at 3kg/1000kg FM, but not at higher dose. Accumulation of Bax was increased in ovaries of rats fed by bee pollen at doses either 3 or 5kg/1000kg FM, whilst accumulation of caspase-3 increased after feeding with bee pollen at dose 5kg/1000kg FM, but not at 3kg/1000kg FM. Our results contribute to new insights regarding the effect of bee pollen on both secretion activity (release of growth factor IGF-I and steroid hormones progesterone and estradiol) and apoptosis (anti- and pro-apoptotic markers Bcl-2, Bax and caspase-3). Bee pollen is shown to be a potent regulator of rat ovarian functions. © 2012 Blackwell Verlag GmbH. Source


Kolesarova A.,Slovak University of Agriculture | Capcarova M.,Slovak University of Agriculture | Sirotkin A.V.,Institute for Genetics and Reproduction of Farm Animals | Medvedova M.,Slovak University of Agriculture | Kovacik J.,Slovak University of Agriculture
Journal of Trace Elements in Medicine and Biology | Year: 2011

The general objective of this in vitro study was to examine the secretory activity (insulin-like growth factor I, IGF-I) of porcine ovarian granulosa cells after Ag addition and to outline the potential intracellular mediators (cyclin B1 and caspase-3) of its effects. Ovarian granulosa cells were incubated with silver nitrate (AgNO 3) at the doses 0.09, 0.17, 0.33, 0.5 and 1.0mg/mL for 18h and compared to the control group without metal addition. The release of IGF-I by granulosa cells was assessed by RIA and expression of cyclin B1 and caspase-3 immunocytochemistry. Our observations show that IGF-I release by granulosa cells was significantly (P<0.05) stimulated by AgNO 3 addition at the doses (0.09-1.0mg/mL). Similarly to IGF-I the cyclin B1 and caspase-3 expression in ovarian granulosa cells was stimulated by Ag addition (0.09-1.0mg/mL). In conclusion, the present results indicate, a direct effect of Ag on (1) secretion of growth factor IGF-I, (2) expression of markers of proliferation (cyclin B1) and apoptosis (caspase-3) of porcine ovarian granulosa cells and (3) that the effect of Ag on ovarian cell proliferation could be mediated by IGF-I and cyclin B1. Obtained data indicate the interference of Ag in the pathways of proliferation and apoptosis of porcine ovarian granulosa cells through hormonal and intracellular peptides such as are cyclin B1 and caspase-3. © 2011 Elsevier GmbH. Source


Kolesarova A.,Slovak University of Agriculture | Capcarova M.,Slovak University of Agriculture | Maruniakova N.,Slovak University of Agriculture | Lukac N.,Slovak University of Agriculture | And 2 more authors.
Journal of Environmental Science and Health - Part A Toxic/Hazardous Substances and Environmental Engineering | Year: 2012

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. Copyright © Taylor & Francis Group, LLC. Source

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