The stimulatory effect of alpha-melanotropin on progesterone release from rat granulosa cells is inhibited by interleukin-1beta and by tumour necrosis factor-alpha.

Abstract

Several studies have shown that a variety of peptides and cytokines are involved in ovarian regulatory mechanisms; however, their exact function is still unclear. In this work we study whether the administration of peptide alpha-melanotropin and the cytokines interleukin-1beta (IL-1beta) and tumour necrosis factor-alpha (TNF-alpha) on their own modify the release of progesterone in cultured granulosa cells (GC) from pro-oestrous rats. We also investigate an interaction between these cytokines and alpha-melanotropin in the modulation of progesterone secretion. Granulosa cells were collected from the ovaries of female Wistar rats and cultured for up to 24 h in the presence of different concentrations of alpha-melanotropin, cytokines or a combination of both. Progesterone concentration was measured by radioimmunoassay. The addition of alpha-melanotropin in a dose of 0.01 and 0.1 mm had no effect on progesterone release, whereas a dose of 1 mm significantly increased progesterone release (P < 0.01) compared with the control culture. Progesterone release was not modified when different concentrations of interleukin-1beta or TNF-alpha were added to the cell cultures. However, when interleukin-1beta or TNF-alpha were added simultaneously with 1 microm alpha-melanotropin, a significant reduction (P < 0.01 for interleukin-1beta and P < 0.05 for TNF-alpha) of the steroid release was found with respect to the alpha-melanotropin-treated group. These results lead us to suggest that, although alpha-melanotropin stimulates progesterone release in pre-ovulatory GC, this effect is blocked by the presence of interleukin-1beta or TNF-alpha.