Are oestrogen receptors and protein tyrosine kinases involved in phytoestrogen-modulated steroid secretion by porcine adrenocortical cells?

The phytoestrogens genistein and daidzein had been found to affect the function of some tissues via oestrogen receptors (ER). In addition, genistein, but not daidzein, is considered to be a protein tyrosine kinase (PTK) inhibitor. Thus, the involvement of oestrogen receptors and PTK in phytoestrogen action on adrenocortical porcine steroidogenesis was examined in this study. The aims of the experiment were to test the effects of (i) ICI 182, 780 (ICI), an ER antagonist, on genistein- and daidzein-modulated cortisol and androstenedione (A4) secretion by adrenocortical cells isolated during the luteal and follicular phases of the porcine oestrous cycle; (ii) tyrphostin AG 957 (TAG), a nonsteroidal PTK inhibitor, on cortisol and A4 secretion by the cells and (iii) the phase of the porcine oestrous cycle on the mechanism of phytoestrogen action. Adrenals were harvested during the luteal (n = 5 animals) and follicular (n = 5 animals) phases of the oestrous cycle from locally slaughtered crossbred gilts. The isolated adrenocortical cells were incubated for 8 h (37 °C, 95% air, 5% CO2) with genistein (5 or 10 µM) or daidzein (5 or 10 µM) in the presence or absence of ICI (0.5 µM) or TAG (5 or 10 µM). Genistein and daidzein inhibited cortisol secretion and stimulated A4 secretion by porcine adrenocortical cells harvested during both the luteal and follicular phases of the oestrous cycle. The ER antagonist ICI did not eliminate phytoestrogen-induced changes in steroidogenesis. In contrast to genistein, TAG reduced the secretion of A4 and did not affect cortisol secretion. There was no observable effect due to the phase of the cycle. It is suggested that the mechanism of genistein and daidzein action in the adrenocortical cells of pigs is independent of ER and PTK. It is possible that PTK are involved in A4 secretion by porcine adrenocortical cells.

Mechanism of phytoestrogen action in Leydig cells of ganders (Anser anser domesticus): Interaction with estrogen receptors and steroidogenic enzymes.

Phytoestrogens (PE) are plant-derived compounds that have an estrogen-like activity and they can influence male and female reproduction. The possible mechanisms of PE action may be including: the binding to estrogen receptors (ER) and the interaction with the key steroidogenic enzymes. The aim of this study was to investigate if PE has effect on steroidogenesis of gander testicular cells by above-described pathways. The Leydig cells were isolated from testes of White Koluda ganders at the peak of their reproductive activity (March). These Leydig cells (1 × 10(5)per mL) were pre-incubated with the ER inhibitor - ICI 182, 780 (100 nM) for 3 h and then these cells were incubated with PE (5 and 50 µM): genistein, daidzein, equol and coumestrol during next 20 h or untreated control and the Leydig cells that were previously treated (20 h) with genistein (5 and 50 µM) were incubated for next 6 h with steroid intermediates (20 µM) as testosterone (T) precursors: hydroxycholesterol, pregnenolone, progesterone and androstenedione. Concentrations of T in the samples of incubation medium were measured using radioimmunoassay. Genistein, daidzein, and equol (5 and 50 µM) decreased (P < 0.05) T secretion by incubated gander Leydig cells and ICI 182, 780 did not eliminate the inhibitory effect of these PE. After genistein (50 µM) treatment, basal and stimulated with 22R-hydroxycholesterol, pregnenolone, progesterone and androstenedione, T production by testicular cells was decreased (P < 0.05). In contrast, genistein at lower dose (5 µM) did not affect the stimulatory effects of testosterone precursors. In conclusion, the inhibition of testosterone secretion by the phytoestrogens in gander Leydig cells did not depend on estrogen receptors. The suppression of steroidogenesis in these cells may be in part conducted by interaction of phytoestrogens with key steroidogenic enzymes. However, further studies are required to elucidate the phytoestrogen mechanism of action in gander testicular cells.

Effects of dietary meals containing different levels of phytoestrogens on reproductive function in Bilgoraj ganders.

The aim of this study was to investigate how long-term feeding of ganders with diets containing different levels of phytoestrogens affects the morphometry of testes, plasma steroid levels, sperm parameters and reproductive performance. Male Bilgoraj geese were fed diets containing grass meal (with low phytoestrogen levels) and those containing alfalfa meal and soybean meal (with higher levels of phytoestrogens). Testes were obtained from ganders at the peak of mating and the ongoing laying period as well as at the beginning of photorefractoriness. The morphometric parameters were measured in frozen testicular sections. Blood was collected monthly from December to June, and plasma testosterone, androstenedione and corticosterone concentrations were determined. During the mating season, semen was collected and its quantity and quality were investigated. In the flocks (ganders with geese) reproductive output was noted. Seasonal changes were observed in the morphology of testes and in plasma testosterone, androstenedione and corticosterone concentrations in ganders. The feeding of ganders with dietary meals containing higher levels of phytoestrogens did not affect the plasma steroid contents or the majority of the morphometric parameters of testes, except for the height of the seminiferous epithelium, which was lowered. Moreover, diets with higher levels of phytoestrogens decreased the volume of ejaculates and the percentage of normal spermatozoa. However, these changes were not reflected in the fertilisation rates.

Phytoestrogens alter cortisol and androstenedione secretion by porcine adrenocortical cells.

The effect of plant-derived, nonsteroidal estrogens (genistein, daidzein and biochanin A) on cortisol and androstenedione secretion by porcine adrenocortical cells was examined. Adrenals were harvested from locally slaughtered mature gilts on days 5-9 of the oestrous cycle. Adrenocortical cells were incubated with or without genistein, daidzein, biochanin A (0.5, 5, 10 or 50 microM), oestradiol (10, 50, 100 or 500 pg/mL) and ACTH (5 nM--positive control). All incubations were performed for 8 h (95% air and 5% CO2, 37 degrees C). ACTH increased cortisol and androstenedione secretion. Genistein, daidzein and biochanin A suppressed cortisol output, whereas androstenedione secretion was enhanced by these phytoestrogens. In contrast, oestradiol did not alter steroid secretion by porcine adrenocortical cells, which may suggest that phytoestrogens have a non-oestrogenic mechanism of action. Therefore, phytoestrogens present in commercial forage may influence adrenocortical function in pigs by decreasing cortisol and increasing androstenedione secretion.

Seasonal changes in the hypothalamo-pituitary-gonadal axis in birds.

Seasonal reproduction in birds is the adaptation to breed at the time of the greatest survival of young. This exact moment is mainly imposed by the photoperiod which stimulates the hypothalamo-pituitary-gonadal axis and starts breeding season. This article summarizes present knowledge concerning: 1/ perception and transduction of light into the biological signal; 2/ model of the avian photoperiodic response; 3/ seasonal changes in hypothalamic secretion of gonadotropin-releasing hormone (GnRH), vasoactive intestinal polipeptide (VIP), as well as the gonadotropin inhibitory hormone (GnIH); 4/ seasonal interactions between pituitary hormones; 5/ seasonal morphological and functional changes within the avian gonads.

Mechanism of phytoestrogens action in reproductive processes of mammals and birds.

Phytoestrogens are polyphenolic compounds that occur ubiquitously in food of plant origin and they have a variety of biological effects in numerous animal cell systems in vivo as well in vitro. Results of studies conducted on animals have shown that effects of phytoestrogens vary depending on species, sex, routes of administration, dose and exposure time. This review summarizes the results of ours studies concerning: 1/ molecular mechanism of phytoestrogen action in porcine granulosa cells, 2/ the involvement of phytoestrogens in immunological regulations of bovine corpus luteum function during luteolysis, 3/ genistein action on metabotropic hormones and lipid-carbohydrate metabolism in rats during pregnancy, 4/ the effects of phytoestrogens on reproductive processes in males of bilgoraj goose.

Phytoestrogen action in Leydig cells of Bilgoraj ganders (Anser anser).

The mechanism of phytoestrogen action in gonadal cells of ganders has not been elucidated. The aim of the study was to investigate in Bilgoraj ganders the possibility of phytoestrogen action via estrogen or androgen receptors or via protein tyrosine kinase pathways in Leydig cells. Genistein and daidzein (5 and 50 microM) as well as equol (50 microM) inhibited testosterone (T) secretion by incubated Leydig cells (1x10(5)/ml; 20 h; 37 degrees C). The effects of hydroxytamoxifen (estrogen receptor inhibitor) and cyproterone acetate (androgen receptor antagonist) on phytoestrogen inhibition of T release by Leydig cells were not observed. Lavendustin A (protein tyrosine kinases inhibitor) did not change T production. The influence of phytoestrogens seems not to be conducted via estrogen and androgen receptors or protein tyrosine kinases system in these cells, but further studies are required to completely examine the mechanism of phytoestrogens action in testes of ganders.

Genistein affects testosterone secretion by Leydig cells in roosters (Gallus gallus domesticus).

Genistein is one of non-steroidal phytoestrogens present in soya and soybean products as well as in other legumes. Phytoestrogens possess estrogen-like biological activity and may influence human and animal reproduction. The aim of this study was to examine the effect of genistein on testosterone (T) secretion by isolated Leydig cells in roosters. Genistein (5-50 microM) inhibited (p<0.05) in vitro basal and LH-stimulated T secretion by Leydig cells in a dose dependent manner. No significant effect of lavendustin C (inhibitor of PTK, a non-phytoestrogen) on the T production was observed. In conclusion, genistein, present in commercial poultry feeds, may influence testicular steroidogenesis but its effect on reproductive performance of roosters requires further examinations.