Ghrelin regulates secretion and proliferation of the bovine oviductepithelial cells in vitro
2010
Fazekašová, J., Slovak Univ. of Agriculture in Nitra (Slovak Republic) | Makarevich, A.V., Animal Production Research Centre, Nitra (Slovak Republic) | Sirotkin, A.V., Animal Production Research Centre, Nitra (Slovak Republic) | Bulla, J., Slovak Univ. of Agriculture in Nitra (Slovak Republic)
The aim of our in vitro experiments was to investigate the role of ghrelin (Ghr), a newly discovered metabolic hormone in control of proliferation (PCNA, cyclin B1), apoptosis (Bax, p53) and secretory activity (IGF-I and PGF 2 alpha release) of bovine oviductal cells in vitro. The cells were isolated by flushing of the oviducts from slaughtered cows. The cells were pre-cultured in a basal medium (DME/F12 medium + 20% FCS + 1% antibiotic-antimycotic solution) to form a monolayer and then were incubated in presence of Ghr at doses of 0, 1, 10 or 100 ng per ml for 72 hours. Concentrations of insulin-like growth factor I (IGF-I) and prostaglandin F2 alpha (PGF2 alpha ) in cell-conditioned medium were measured by IRMA/RIA kits. Indices of cell apoptosis (apoptotic peptides Bax and p53) or proliferation (the proliferation associated peptides PCNA and cyclin B1) in the bovine oviduct epithelial cells were analyzed by immunocytochemistry (ICC) and Western-blotting (WB). Obtained results show that Ghr stimulated the output of PGF 2 alpha (at all tested doses; p less than 0.05) and IGF-I (at 1 and 100 ng per ml; p less than 0.05) by the oviductal cells. Ghr, given at all doses, increased the proportion of PCNA-positive cells (ICC, WB). However, the proportion of Bax-, cyclin B1- and p53- positive cells (ICC) was not changed significantly after addition of Ghr. These results clearly demonstrate that Ghr is a potent regulator of the secretory activity and proliferation in the oviduct epithelial cells. Elucidation of ghrelin role in apoptosis regulation within the oviduct requires further investigations.
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