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prof. Ing. Adriana Kolesárová, PhD.
Identifikačné číslo: 1505
Univerzitný e-mail: adriana.kolesarova [at] uniag.sk
 
profesorka CSc./PhD. - Katedra fyziológie živočíchov (FBP)
dekanát - centrum - Fakulta biotechnológie a potravinárstva
Prodekanka - Fakulta biotechnológie a potravinárstva

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Typ práce: Dizertačná práca
Názov práce:Toxické aspekty trichotecénov v ovariálnych bunkách in vitro
Autor: Ing. Nora Maruniaková, PhD.
Pracovisko: Katedra fyziológie živočíchov (FBP)
Vedúci práce: prof. Ing. Adriana Kolesárová, PhD.
Oponent 1:prof. Ing. Róbert Toman, Dr.
Oponent 2:Ing. Alexander Makarevič, DrSc.
Oponent 3:prof. Ángel Carbonell Barrachina
Stav záverečnej práce:Záverečná práca bola úspešne obhájená


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Jazyk spracovania záverečnej práce:anglický jazyk

slovenský jazyk        anglický jazyk

Názov práce:Toxic aspects of trichothecenes on ovarian cells in vitro
Abstrakt:T-2 toxin and its metabolite HT-2 toxin are one of the most toxic mycotoxins of type A trichothecenes which causes a different toxic effect in both animal and human. They may reduce reproductive performance in animals and influence functions of endocrine and reproductive tract. The objective of this in vitro study was to examine changes of secretion activity (RIA and ELISA method) of ovarian cells to produce progesterone (P4) and estradiol (E2) and to outline parameters of cell proliferation and apoptosis (imunocytochemistry, westernblotting method). In porcine ovarian granulosa cells (GCs) we determined secretion activity after addition of a) alone T-2 toxin and HT-2 toxin, b) different time duration of these toxins 12, 24, 48 hours long treatment, c) combination with various substances (growth factor IGF-I, hormones leptin and ghrelin). In rabbit ovarian cells we examined secretion activity after d) alone T-2 toxin and HT-2 toxin e) combination with various substances (growth factor IGF-I, hormones leptin and ghrelin) finally we examined f) markers of proliferation and apoptosis. All experimental groups are compared to control group. We observed significant (P<0.05) stimulation in P4 release by ovarian GCs of non-cyclic gilts after addition of 1000 ng.ml-1 of T-2 toxin and 100, 1000 ng.ml-1 of HT-2 toxin. E2 release no significantly decreased after addition of all used doses of both toxins. In experiments with ovarian GCs of cyclic gilts we observed significant (P<0.05) stimulation in P4 release after addition of T-2 toxin at doses 1 and 10 ng.ml-1 and 1, 10, 100 ng.ml-1 of HT-2 toxin. E2 release by cyclic GCs of gilts was significantly (P<0.05) stimulated after T-2 toxin at 10 ng.ml-1 and 100 ng.ml-1. Experiments with combined effect of toxins showed significant (P<0.05) stimulation in P4 release after addition of 1, 100 ng.ml-1 of T-2 toxin with HT-2 toxin. After 12 hours long treatment of ovarian GCs of gilts with 100 ng.ml-1 of HT-2 toxin we observed significant (P<0.05) stimulation in P4 release. After 24 hours long treatment of ovarian GCs of gilts by T-2 toxin at 1, 10 ng.ml-1 we observed significant (P<0.05) inhibition in P4 release. HT-2 toxin caused significant (P<0.05) stimulation in P4 release at 10 ng.ml-1 of this toxin to ovarian GCs. After 48 hours long treatments of T-2 toxin significant (P<0.05) stimulation was observed at highest used dose 100 ng.ml-1. In experiments with combined effect of toxins with IGF-I, ghrelin and leptin we observed significant (P<0.05) inhibition of P4 release after addition at all used doses of of T-2 toxin (10, 100, 1000 ng.ml-1) with IGF-I (100 ng.ml-1). Similarly addition of combined effect of HT-2 toxin with IGF-I we observed significant (P<0.05) inhibition at (10, 100 ng.ml-1) of toxin plus 100 ng.ml-1 of IGF-I. In experiment with combined effect of toxins with leptin we observed significant (P<0.05) inhibition after addition 100 ng.ml-1 of T-2 toxin with 1000 ng.ml-1 of leptin. Compare to control group we detected significant (P<0.05) stimulation in addition of 1000 ng.ml-1 of HT-2 toxin with 1000 ng.ml-1 of leptin. Combination of T-2 toxin at 10 ng.ml-1 with 500 ng.ml-1 of ghrelin inhibited P4 release by GCs. T-2 toxin and HT-2 toxin caused no significant changes in P4 release by ovarian cells of rabbits. E2 release by rabbit ovarian fragments was significantly (P<0.05) inhibited after addition of 0.01, 10, 100 ng.ml-1 of HT-2 toxin. P4 and E2 release was significantly (P<0.05) inhibited after addition of all used combined doses of T-2 toxin (0.01, 0.1, 1, 10, 100 ng.ml-1) with IGF-I (100 ng.ml-1). E2 release by rabbit ovarian cells was significantly (P<0.05) inhibited after addition of T-2 toxin (10, 100 ng.ml-1) with leptin (1000 ng.ml-1). The percentage of ovarian GCs containing PCNA was significantly (P<0.05) higher after (0.01, 1, 10, 100 ng.ml-1) of T-2 toxin and significantly decreased (P<0.05) after HT-2 toxin (1, 10, 100 ng.ml-1) during 24 hours long treatment. After 12 hours the percentage of ovarian GCs containing cyclin B1 was significantly (P<0.05) higher at 100 ng.ml-1 of HT-2 toxin. The percentage of ovarian GCs containing cyclin B1 significantly (P<0.05) increased after 24 hours long treatment of T-2 toxin at doses 1 and 100 ng.ml-1 and after HT-2 toxin at 100 ng.ml-1. After 24 hours the percentage of ovarian GCs containing caspase-3 significantly (P<0.05) decreased after addition of HT-2 toxin at doses 0.1 and 10 ng.ml-1. The percentage of ovarian GCs containing p-53 significantly (P<0.05) decreased after T-2 toxin at 1 ng.ml-1 after 12 hours long treatment. In conclussion, the results of our in vitro experiments indicate the possible time and dose-dependent effect of T-2 toxin and HT-2 toxin on secretion activity of ovarian GCs as well as up-regulation or down-regulation of proliferation and apoptosis by certain markers. Currently, the study of mycotoxins and their impact on animal health is very required issue because of their frequently occurence in various agricultural commodities and feeds.
Kľúčové slová:HT-2 toxin, T-2 toxin, steroid hormones

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