In vitro Assessment of Iron on Porcine Ovarian Granulosa Cells: secretory activity, markers of proliferation and apoptosis

SUMMARY 14 15 The obtained data could expand the existing general knowledge concerning direct action of 16 metals on the ovary. Nevertheless, the results of testing of iron compound on porcine 17 ovarian cells should be interpreted carefully because iron is an essential element but on the 18 other hand it could induce changes in cellular processes. The general objective of this in 19 vitro study was to first examine dose–dependent effects of iron on the secretory activity of 20 porcine ovarian granulosa cells, secondly to outline the potential intracellular mediators that 21 mediate these effects. Specifically, we evaluated the effect of FeSO 4. 7H 2 O on the release of 22 insulin-like growth factor I (IGF-I) and progesterone, as well as the expression of markers 23 of proliferation (cyclin B1) and apoptosis (caspase-3) in porcine ovarian granulosa cells. 24 Concentrations of IGF–I and progesterone were determined by RIA, cyclin B1 and caspase-25 3 expression by immunocytochemistry (ICC). Our results show a significantly decreased 26 IGF–I secretion by ovarian granulosa cells after Fe addition at the doses 0.5 and 1.0 mg/ml. 27 The iron additions at doses 0.17 and 1.0 mg/ml had no effect on progesterone secretion. In 28 contrast, iron addition at doses 0.17 and 1.0 mg/ml, resulted in stimulation of cyclin B1 and 29 caspase-3 expression. In conclusion, the present results indicate, (1) a direct effect of Fe on 30 secretion of growth factor IGF-I but not steroid hormone progesterone, (2) expression of 31 markers of proliferation (cyclin B1) and (3) apoptosis (caspase-3) of porcine ovarian 32 granulosa cells. These results support an idea that iron could play a regulatory role in 33 porcine ovarian function: hormone release, proliferation and apoptosis. 34 35