Interleukin-1α and -β modulation of luteinized human granulosa cell oestrogen and progesterone biosynthesis

Abstract

This study was designed to test the hypothesis that interleukin-1 alpha (IL-1 alpha) and beta directly affect progesterone, and oestradiol production in cultures of purified human granulosa cells. Luteinized granulosa cells were obtained from women during in-vitro fertilization cycles. Granulosa cells with and without associated white blood cells were cultured in the presence of IL-1 alpha and IL-1 beta (0.5-50 ng/ml) for 48 h. Media were changed at 24 h intervals and assayed for progesterone and oestradiol. In separate experiments, granulosa cell viability was assessed with the tetrazolium salt reduction assay, haemocytometer cell counts, and Trypan blue dye exclusion. Our results indicate that progesterone synthesis by basal and human chorionic gonadotrophin (HCG)-stimulated granulosa cells co-cultured with white blood cells was inhibited by 5.0 ng/ml of IL-1 alpha and IL-1 beta at 48 h of culture. In the presence of white blood cells, granulosa cell oestradiol synthesis was inhibited by IL-1 beta but not IL-1 alpha. Oestradiol was inhibited after both 24 and 48 h of culture and was maximally affected by 5.0 ng/ml of IL-1 beta. In contrast, basal and HCG-stimulated oestradiol production by granulosa cells cultured free of white blood cells was inhibited only by IL-1 alpha. IL-1 alpha at 5.0 ng/ml produced maximal inhibition of basal oestradiol (57%) and HCG-stimulated oestradiol (41%) production at 48 h of culture. Gonadal steroid inhibition by IL-1 alpha and IL-1 beta was not mediated through cytotoxic or antiproliferative effects on granulosa cells. Specificity of the granulosa cell response to IL-1 alpha and IL-1 beta was demonstrated by abrogation of steroid inhibition with anti-IL-1 alpha and IL-1 beta neutralizing antibodies. In conclusion, IL-1 alpha directly inhibited the production of oestradiol by human ovarian granulosa cells. IL-1 alpha and IL-1 beta also exerted indirect effects on steroid production via white blood cells that are usually present in granulosa cell cultures if steps are not taken to remove them. These data support the hypothesis that cytokines play an important role in intra-ovarian regulation of steroid biosynthesis.

High doses of dietary zinc induce cytokines, chemokines, and apoptosis in reproductive tissues during regression

“…In our earlier studies, we have noticed a reduction of estrogen and progesterone in birds stimulated to molt by high doses of zinc (Anish et al 2007). In mammals, proinflammatory cytokines have been found to inhibit ovarian steroidogenesis by inhibiting the gonadotropin-support (Bornstein et al 2004;Breard et al 1998aBreard et al , 1998bHurwitz et al 1991;Best and Hill 1995;Yasuda et al 1990;Young et al 1997). Therefore, the effect of zinc on ovarian steroids might be mediated through cytokines.…”

Effects of acute psychological stress on adhesion molecules, interleukins and sex hormones: implications for coronary heart disease

“…IL-1 application, on the other hand, inhibits estradiol production (Best and Hill 1995). Since the IL-1 dependent induction of vascular cell adhesion molecules was inhibited by estradiol (Caulin-Glaser et al 1996), stress-induced reductions in estradiol concentrations may disinhibit the expression of cell adhesion molecules.…”

Polycystic ovary syndrome and circulating inflammatory markers

“…Interleukin-1 alpha and beta directly affect progesterone and oestradiol production in cultures of purified human granulosa cells (17). IL-1 alpha directly inhibites the production of oestradiol by human ovarian granulosa cells (18). IL-1β stimulates basal progesterone secretion in the human granulosa and theca cells and in small and large follicles in vitro (19).…”