The effect of deoxynivalenol on the secretion activity, proliferation and apoptosis of porcine ovarian granulosa cells in vitro

Abstract

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.

Keywords:

• Deoxynivalenol
• Insulin-like growth factor
• progesterone
• proliferation
• apoptosis
• granulosa cell

Acknowledgments

The authors are thankful to Romer Labs Division Holding GmbH, Tulln, Austria for providing mycotoxins and colleagues from the Institute for Genetics and Reproduction of Farm Animals, Animal Production Research Centre Nitra, particularly to Mrs. Katarina Tothova and Ing. Zofia Kuklova for their skillful technical assistance and MSc. S. Roychoudhury, PhD. for English linguistic revision. This work was financially supported by the Slovak Research and Development Agency project nos. 0299–06, the Ministry of Education of the Slovak Republic project nos. 1/0790/11, 1/0696/08 and 1/0834/08.