A potential physiological involvement of nesfatin-1 in regulating swine granulosa cell functions

Nesfatin-1 has recently been indicated as a pleiotropic molecule, mainly involved in the metabolic regulation of reproductive functions acting at hypothalamic level. The aim of the present study was to explore a local action of nesfatin-1 in swine ovarian follicle. Firstly, the expression of NUCB-2 by qRT-PCR was verified in swine granulosa cells from different size follicles and nesfatin-1 was localized by immunohistochemistry in sections of the whole porcine ovary. In addition, the in vitro effect of different nesfatin-1 concentrations was tested on cell growth, steroidogenesis and redox status of granulosa cells. Finally, the effect of nesfatin-1 was also studied in an angiogenesis bioassay since vessel growth is essential for ovarian follicle function. Immunohistochemistry revealed intense positivity for nesfatin-1 in swine granulosa cells in all developmental stage follicles. The precursor protein nucleobindin-2 (NUCB-2) showed a higher expression in granulosa cells from large follicles compared to medium and small follicles. In addition, our results demonstrated that nesfatin-1 stimulates cell proliferation and progesterone production and interferes with redox status by modifying NO production and non enzymatic scavenging activity in granulosa cells from large follicles. Moreover, nesfatin-1 displays a stimulatory effects on angiogenesis. Present study demonstrates, for the first time, that nesfatin-1 is physiologically present in the swine ovarian follicle where it could impair granulosa cell functions.