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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 | And 3 more authors.
Journal of Environmental Science and Health - Part A Toxic/Hazardous Substances and Environmental Engineering | Year: 2011

Molybdenum (Mo) is an essential trace element and it plays an important role in cell functions. The mechanism of the action of molybdenumin connection with growth factor IGF-I, proliferation-related peptide cyclin B1 and apoptosis-related peptide caspase-3 has not been examined previously in porcine ovarian granulosa cells. The general objective of this in vitro study was to examine the secretory activity of porcine ovarian granulosa cells after experimental Mo administration and to outline the potential intracellular mediators of its effects. Ovarian granulosa cells were incubated with ammonium molybdate for 18 hours: 1.0 mg/ml; 0.5 mg/mL; 0.33 mg/mL; 0.17 mg/mL and 0.09 mg/mL, while the control group received no Mo. The secretion of IGF-I was assessed by RIA and expression of cyclin B1 and caspase-3 by immunocytochemistry. IGF-I release was decreased by Mo addition at the doses 1.0 mg/mL and 0.5 mg/mL. The expression of cyclin B1 was stimulated by Mo addition at all doses ranging from 1.0 - 0.09 mg/mL. Caspase-3 expression was also stimulated after experimentalMo addition at the doses 1.0 and 0.5 mg/mL. These data contribute to new insights regarding the mechanism of action of Mo on porcine ovarian functions, secretory activity, proliferation and apoptosis of granulosa cells through hormonal and intracellular substances such as are cyclin B1 and caspase-3. Copyright © Taylor & Francis Group, LLC.


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.


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.


Kolesarova A.,Slovak University of Agriculture | Capcarova M.,Slovak University of Agriculture | Sirotkin A.,Institute for Genetics and Reproduction of Farm Animals | Medvedova M.,Slovak University of Agriculture | Kovacik J.,Slovak University of Agriculture
Journal of Environmental Science and Health - Part A Toxic/Hazardous Substances and Environmental Engineering | Year: 2010

Cobalt (Co) is an essential element. The general objective of this in vitro study was to examine dose-dependent changes in the secretory activity of porcine ovarian granulosa cells after experimental Co administration and to outline the potential intracellular mediators of its effects. Concentrations of IGF-I and progesterone were determined by RIA and expression of cyclin B1 and caspase-3 by immunocytochemistry. IGF-I release by granulosa cells was stopped by Co addition at the concentration 1 mg/mL. Progesterone release by granulosa cells was decreased at the lowest Co addition (0.09 mg/mL). In our study the changes of the expression of proliferation related peptide cyclin B1 and apoptosis related peptide caspase-3 in ovarian granulosa cells was observed after experimental Co addition. The molecular pathways stimulated by Co through the expression of cyclin B1 and caspase-3 were found. In conclusion, this study provides novel evidence that cobalt is the factor which can initiate adverse effects in ovarian granulosa cells. These results contribute towards the understanding of mechanisms relating to endocrine disruptor-induced alterations in porcine ovarian granulosa cells. Copyright © Taylor & Francis Group, LLC.


Marina M.,Slovak University of Agriculture | Adriana K.,Slovak University of Agriculture | Marcela C.,Slovak University of Agriculture | Roman L.,Romer Labs Division Holding GmbH | And 3 more authors.
Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural Wastes | Year: 2011

The aim of this in vitro study was to examine the secretion activity, markers of proliferation and apoptosis in porcine ovarian granulosa cells (GCs) after deoxynivalenol (DON) addition. Ovarian granulosa cells were incubated with DON for 24h: 10, 100 and 1000 ng/mL, while the control group received no DON. The secretion of insulin-like growth factor I (IGF-I) and progesterone was determined by radioimmunoassay (RIA) and expression of cyclin B1, PCNA and caspase-3 by immunocytochemistry. IGF-I release by GCs was inhibited by DON, while progesterone release and the expression of cyclin B1 was stimulated by DON (at 1000 ng/mL but not at 10 and 100 ng/mL). PCNA expression was stimulated by DON (at 100 and 1000 ng/mL but not at 10 ng/mL). Caspase-3 expression was not influenced by DON treatment (at all doses). In conclusion, our results indicate, (1) a direct effect of DON on secretion of growth factor IGF-I and steroid hormone progesterone, (2) expression of markers of proliferation (cyclin B1 and PCNA) but not on the (3) expression of marker of apoptosis (caspase-3) in porcine ovarian granulosa cells. This in vitro study suggests the dose-dependent association of DON on porcine ovarian functions. ©Taylor & Francis Group, LLC.


Adriana K.,Slovak University of Agriculture | Capcarova M.,Slovak University of Agriculture | Bakova Z.,Slovak University of Agriculture | Branislav G.,Slovak University of Agriculture | And 3 more authors.
Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural Wastes | Year: 2011

The general objective of this in vitro study was to examine the effect of bee pollen on the release of insulin-like growth factor I (IGF-I) and steroid hormone progesterone, and expression of markers of proliferation (PCNA) and apoptosis (caspase-3) in porcine ovarian granulosa cells. Concentrations of IGF-I and progesterone were determined by RIA method and expression of PCNA and caspase-3 by immunocytochemistry. Bee pollen addition at the dose of 10 ng/mL significantly (P < 0.05) inhibited IGF-I release by porcine ovarian granulosa cells. This growth factor was not influenced by 100 and 1000 ng/mL doses of bee pollen. Progesterone release by cells was not influenced by bee pollen addition at the doses of 10, 100 and 1000 ng/mL as used in our study. Similarly expression of PCNA and caspase-3 was not affected by bee pollen addition. The present study shows dose-dependent regulation of IGF-I by experimental bee pollen addition in vitro. Progesterone release, expression of PCNA and caspase-3 in porcine ovarian granulosa cells was not induced by pollen. Our results contribute to new insights regarding the possible effect of bee pollen on IGF-I release, which is important for regulation of porcine ovarian functions. © Taylor & Francis Group, LLC.


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.


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.


Maruniakova N.,Slovak University of Agriculture | Kadasi A.,Slovak University of Agriculture | Kadasi A.,Institute for Genetics and Reproduction of Farm Animals | Sirotkin A.V.,Institute for Genetics and Reproduction of Farm Animals | And 2 more authors.
Journal of Environmental Science and Health - Part A Toxic/Hazardous Substances and Environmental Engineering | Year: 2014

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, occurence 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. © 2014 Copyright Taylor and Francis Group, LLC.


PubMed | Institute of Animal Science, Institute for Genetics and Reproduction of Farm Animals, Assam University and Slovak University of Agriculture
Type: Journal Article | Journal: International journal of impotence research | Year: 2015

Ricinus communis L. has ethnopharmacological contraceptive reputation but its stem bark has unexplored mechanisms of action in female reproductive system. In the present study, the effect of methanolic and aqueous extracts from the stem bark of the plant was examined on basic porcine ovarian granulosa cell functions and its response to Luteinising hormone (LH)-the upstream hormonal regulator. Systemic treatment of methanolic and aqueous extracts stimulated cell proliferation (proliferating cell nuclear antigen, PCNA) and also promoted cell apoptosis (caspase-3). Aqueous extract has inverted the stimulatory effect of LH on PCNA but not on caspase-3. Methanolic extract stimulated as well as inhibited progesterone release and stimulated testosterone secretion. Whereas aqueous extract inhibited both steroid releases and suppressed the stimulatory effect of LH on progesterone release and promoted the inhibitory effect of LH on testosterone release. In conclusion, the present study unveils the mechanism of action of R. communis stem bark in in vitro condition. These suggest its possible contraceptive efficacy by exerting its regulatory role over LH and on basic ovarian cell functions and secretion activity.

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