RESUMEN
This review is to summarize and analyze the currently available knowledge concerning the action of oat (Avena sativa L.) consumption on obesity, as well as possible constituents and extra- and intracellular mediators responsible for its anti-obesity effect. The oat constituents could reduce fat storage via several mediatory mechanisms - brain centers regulating appetite, gastrointestinal functions, gut bacteria, fat synthesis and metabolism and maybe via changes in oxidative processes, steroid hormones receptors and adipose tissue vascularization. Several oat constituents (starch, fiber and beta-glucan) could have anti-obesity properties, whilst one oat constituent (starch or fiber) could affect fat storage via several mechanisms of action.
Asunto(s)
Avena , Fibras de la Dieta , Humanos , Avena/metabolismo , Fibras de la Dieta/metabolismo , Fibras de la Dieta/farmacología , Obesidad/metabolismo , Tejido Adiposo/metabolismo , Almidón/metabolismo , Almidón/farmacologíaRESUMEN
The study aimed to evaluate the involvement of apigenin, microRNA (miR)-152, and their interrelationships in the control of basic ovarian granulosa cell functions. The effects of apigenin (0, 10, and 100 µg/mL), miR-152 analogues or miR-152 inhibitor, and their combinations with apigenin on porcine granulosa cells were examined. Expression levels of miR-152, viability, proliferation, apoptosis, steroid hormones, IGF-I, oxytocin, and prostaglandin E2 release were analyzed. Apigenin increased the expression of miR-152, cell proliferation, and estradiol release and reduced apoptosis, progesterone, and IGF-I output. MicroRNA-152 analogues promoted cell viability and proliferation, as well as the release of progesterone, IGF-I, oxytocin, and prostaglandin E2; however, it inhibited apoptosis and estradiol output. miR-152 inhibitor had the opposite effect. Moreover, miR-152 analogues suppressed the effect of apigenin on cell apoptosis and estradiol release. These observations 1) confirm the involvement of apigenin in the control of basic ovarian cell functions; 2) are the first demonstration of importance of miR-152 in the control of these functions; 3) show the ability of apigenin to promote miR-152 expression and the ability of miR-152 to modify apigenin effects on ovarian cells.
Asunto(s)
MicroARNs , Progesterona , Femenino , Porcinos , Animales , Progesterona/farmacología , Progesterona/metabolismo , Factor I del Crecimiento Similar a la Insulina/metabolismo , Apigenina/farmacología , Apigenina/metabolismo , Oxitocina/farmacología , Dinoprostona/farmacología , Células Cultivadas , Células de la Granulosa/fisiología , Estradiol/farmacología , Estradiol/metabolismo , Proliferación Celular , Apoptosis , MicroARNs/genética , MicroARNs/metabolismoRESUMEN
The present study examined the effect of medicinal plants - ginkgo, tribulus (puncture vine), and yucca - on ovarian functions and their response to the toxic influence of toluene. Therefore, we analyzed the effect of toluene with and without these plant extracts on cultured human ovarian granulosa cells. Cell viability and the release of progesterone, insulin-like growth factor I (IGF I), oxytocin, and prostaglandin F (PGF) were analyzed using the trypan blue test, enzyme immunoassay, and enzyme-linked immunosorbent assay, respectively. The ginkgo, tribulus and yucca were able to suppress ovarian cell viability and alter the release of hormones. Toluene suppressed cell viability and the release of PGF, but not of progesterone, IGF-I, or oxytocin. The negative effect of toluene on cell viability was prevented and even reversed by ginkgo and yucca, whereas its effect on PGF was prevented or inverted by all tested plant extracts. These findings (1) demonstrated the direct toxic effect of toluene on ovarian cells, (2) showed the direct effect of some medicinal plants on ovarian cell functions, and (3) demonstrated the ability of these plants to inhibit the effects of toluene and to act as natural protectors against the suppressive effect of toluene on female reproduction.
Asunto(s)
Plantas Medicinales , Femenino , Humanos , Oxitocina , Supervivencia Celular , Progesterona , Extractos Vegetales/farmacologíaRESUMEN
Rooibos (Aspalathus linearis Brum. f) can directly influence female reproduction, but whether rooibos can influence the response of ovarian cells to FSH and whether the rooibos effects are due to the presence of quercetin remain unknown. We compared the influence of rooibos extract and quercetin (both at 10 µg/ml-1) on porcine ovarian granulosa cells cultured with and without FSH (0, 1, 10 or 100 ng/ml-1). The expression of intracellular proliferation (PCNA, cyclin B1) and apoptosis (bax, caspase 3) markers in the cells was detected by immunocytochemistry. The release of progesterone (P), testosterone (T) and estradiol (E) were evaluated with ELISAs. Administration of both rooibos and quercetin reduced the accumulation of proliferation markers and promoted the accumulation of apoptosis markers and the release of T and E. Rooibos stimulated, but quercetin inhibited, P output. Administration of FSH increased the accumulation of proliferation markers, decreased the accumulation of apoptosis markers, promoted the release of P and T, and had a biphasic effect on E output. The addition of both rooibos and quercetin mitigated or prevented the main effects of FSH. The present observations suggest a direct influence of both rooibos and quercetin on basic ovarian functions - proliferation, apoptosis, steroidogenesis and response to FSH. The similarity in the major effects of rooibos and its constituent quercetin indicates that quercetin could be the molecule responsible for the main rooibos effects on the ovary. The potential anti-reproductive effects of rooibos and rooibos constituent quercetin, should be taken into account in animal and human nutrition.
Asunto(s)
Aspalathus , Ovario , Humanos , Femenino , Animales , Porcinos , Quercetina/farmacología , Estradiol/farmacología , Hormona Folículo EstimulanteRESUMEN
The present in vitro experiments aimed to examine the effects of the plant polyphenol quercetin and the environmental contaminant toluene on basic ovarian cell functions, including the ability of quercetin to be a natural protector against the adverse effects of toluene. The influence of toluene, quercetin, and their combination on proliferation (accumulation of PCNA), apoptosis (accumulation of bax) and release of progesterone, testosterone and insulin-like growth factor I (IGFI) by cultured porcine ovarian granulosa cells was investigated. Toluene stimulated cell proliferation and inhibited progesterone, IGF-I and testosterone release but did not affect apoptosis. Quercetin, when administered alone, inhibited cell proliferation, apoptosis, IGF-I and testosterone release and stimulated progesterone output. When administered in combination with toluene, quercetin mitigated toluene's effects on proliferation and on progesterone release and induced toluene to exhibit a pro-apoptotic effect. These observations demonstrate the direct effects of both quercetin and toluene on basic ovarian functions and a protective effect of quercetin against the effects of toluene. Therefore, quercetin-containing plants could be regulators of porcine reproduction and natural protectors against the adverse effects of the environmental contaminant toluene.
Asunto(s)
Progesterona , Quercetina , Femenino , Porcinos , Animales , Progesterona/farmacología , Quercetina/farmacología , Factor I del Crecimiento Similar a la Insulina/metabolismo , Tolueno/toxicidad , Tolueno/metabolismo , Células Cultivadas , Células de la Granulosa , Proliferación Celular , Testosterona/metabolismo , ApoptosisRESUMEN
The action of the medicinal plant Tribulus terrestris (TT) on bovine ovarian cell functions, as well as the protective potential of TT against xylene (X) action, remain unknown. The aim of the present in vitro study was to elucidate the influence of TT, X and their combination on basic bovine ovarian cell functions. For this purpose, we examined the effect of TT (at doses of 0, 1, 10, and 100 ng/mL), X (at 20 ?g/mL) and the combination of TT + X (at these doses) on proliferation, apoptosis and hormone release by cultured bovine ovarian granulosa cells. Markers of proliferation (accumulation of PCNA), apoptosis (accumulation of Bax) and the release of hormones (progesterone, testosterone and insulin-like growth factor I, IGF-I) were analyzed by quantitative immunocytochemistry and RIA, respectively. TT addition was able to stimulate proliferation and testosterone release and inhibit apoptosis and progesterone output. The addition of X alone stimulated proliferation, apoptosis and IGF-I release and inhibited progesterone and testosterone release by ovarian cells. TT was able to modify X effects: it prevented the antiproliferative effect of X, induced the proapoptotic action of X, and promoted X action on progesterone but not testosterone or IGF-I release. Taken together, our observations represent the first demonstration that TT can be a promoter of ovarian cell functions (a stimulator of proliferation and a suppressor of apoptosis) and a regulator of ovarian steroidogenesis. X can increase ovarian cell proliferation and IGF-I release and inhibit ovarian steroidogenesis. These effects could explain its anti-reproductive and cancer actions. The ability of TT to modify X action on proliferation and apoptosis indicates that TT might be a natural protector against some ovarian cell disorders associated with X action on proliferation and apoptosis, but it can also promote its adverse effects on progesterone release.
Asunto(s)
Tribulus , Animales , Apoptosis , Bovinos , Proliferación Celular , Células Cultivadas , Femenino , Células de la Granulosa , Factor I del Crecimiento Similar a la Insulina/metabolismo , Progesterona/metabolismo , Testosterona/metabolismo , Tribulus/metabolismo , Xilenos/metabolismo , Xilenos/farmacologíaRESUMEN
The objective of this study was to examine the direct effects of the medicinal plant fennel (Foeniculumvulgare Mill.) on basic functions of ovarian cells, including proliferation, apoptosis, and response to the physiological hormonal stimulator, ghrelin. In the first series of experiments, porcine ovarian granulosa cells were cultured with (1, 10, 100 µg/ml) or without fennel extract. In the second series of experiments, cells were cultured with (1, 10, 100 ng/ml) or without ghrelin, alone or in combination with fennel extract (10 µg/ml). Expression of the proliferation marker, PCNA, and the apoptosis marker, bax, were analyzed via quantitative immunocytochemical methods. Fennel stimulated the accumulation of the proliferation marker, and suppressed the expression of the apoptosis marker. Ghrelin alone promoted proliferation and apoptosis of ovarian cells. The presence of fennel inhibited these ghrelin effects. These observations provide the first demonstration of (1) effects of fennel on farm animal reproduction, (2) direct effects of fennel on ovarian cells, (3) the ability of fennel to promote ovarian cell proliferation, to inhibit ovarian cell apoptosis, and to enhance the ovarian cell proliferation:apoptosis ratio. Furthermore, our results (4) confirm the involvement of ghrelin in the control of ovarian cell apoptosis and proliferation, and (5) demonstrate the ability of fennel to affect not only ovarian cell proliferation and apoptosis, but also to suppress the responses of ovarian cells to the upstream hormonal regulator ghrelin. Our results indicate the potential applicability of fennel as a bio-stimulator of farm animal reproduction.
Asunto(s)
Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Foeniculum , Ghrelina/farmacología , Células de la Granulosa/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Células Cultivadas , Femenino , Foeniculum/química , Células de la Granulosa/metabolismo , Células de la Granulosa/patología , Extractos Vegetales/aislamiento & purificación , Antígeno Nuclear de Célula en Proliferación/metabolismo , Sus scrofa , Proteína X Asociada a bcl-2/metabolismoRESUMEN
This paper reviews provenance, chemical composition and properties of tea (Camelia sinensis L.) and coffee (Coffee arabica, L. and Coffeacaniphora, L.), their general health effects, as well as the currently available knowledge concerning their action on fat storage, physiological mechanisms of their effects, as well as their safety and recommended dosage for treatment of obesity. Both tea and coffee possess the ability to promote health and to prevent, to mitigate and to treat numerous disorders. This ability can be partially due to presence of caffeine in both plants. Further physiological and medicinal effects could be explained by other molecules (theaflavins, catechins, their metabolites and polyphenols in tea and polyphenol chlorogenic acid in coffee). These plants and plant molecules can be efficient for prevention and treatment of numerous metabolic disorders including metabolic syndrome, cardiovascular diseases, type 2 diabetes and obesity. Both plants and their constituents can reduce fat storage through suppression of adipocyte functions, and support of gut microbiota. In addition, tea can prevent obesity via reduction of appetite, food consumption and food absorption in gastrointestinal system and through the changes in fat metabolism.
Asunto(s)
Fármacos Antiobesidad/administración & dosificación , Café , Estado de Salud , Obesidad/prevención & control , Fitoquímicos/administración & dosificación , Té , Adiposidad/efectos de los fármacos , Animales , Fármacos Antiobesidad/efectos adversos , Regulación del Apetito/efectos de los fármacos , Café/efectos adversos , Humanos , Metabolismo de los Lípidos/efectos de los fármacos , Obesidad/diagnóstico , Obesidad/fisiopatología , Fitoquímicos/efectos adversos , Té/efectos adversos , Aumento de Peso/efectos de los fármacosRESUMEN
Tribulus terrestris, L. (puncture vine) have been used as a folk medicine for five thousands of years, but its targets, effects, their mechanisms and application requires further studies. This paper reviews the provenance, constituents and properties of Tribulus terrestris, L., its general physiological and health effects, as well as the currently available knowledge concerning its influence on male and female reproductive processes and their dysfunctions. Analysis of the available publications demonstrated the influence of Tribulus terrestris on a wide spectrum of targets and physiological processe and disorders. In particular, Tribulus terrestris can be a stimulator of male and female reproductive processes at the level of central nervous system, sexual behaviour, pituitary and gonadal hormones and their receptors, gonadal functions (including ovarian follicullogenesis and spermatogenesis), improvement of the quality and quantity of gametes (at least of sperm) and fecundity. This ability of puncture vine is applicable for the improvement of man's sexual desire and sperm quality in vivo and in vitro, as well as of women's libido, activation of women's reproductive organs, fecundity, and treatment of infertility, especially that related to the polycystic ovarian syndrome.
Asunto(s)
Tribulus , Femenino , Humanos , Libido , Masculino , Extractos Vegetales , Reproducción , Conducta SexualRESUMEN
The aim of the present study was to examine the effect of high ambient temperature on rabbit feed consumption, growth, viability, and fecundity, as well as the morphology and endocrine function of gonadal and adrenal cells. Adult does and their offspring were kept at either a comfortable (20°C; control) or high (36°C) temperature throughout pregnancy and up until weaning of pups. Doe mortality and fecundity, and plasma concentrations of hormones were evaluated. In addition, granulosa cells were cultured with and without FSH to assess progesterone production. In the offspring, we assessed mortality, total feed consumption, feed efficiency, growth, plasma hormone concentrations, as well as the microstructure in ovarian granulosa cells, testicular Leydig cells, and adrenocortical cells. We observed greater mortality of both adult animals and offspring at the higher ambient temperature compared with the control. The higher ambient temperature suppressed feed consumption, feed efficiency, and growth of pups. Adult and young females exposed to a high temperature had lower circulating concentrations of progesterone, but not of estradiol, compared with controls. Young males exposed to a high ambient temperature had greater circulating concentrations of testosterone, but not progesterone, compared with controls. High ambient temperature reduced circulating IGF-I concentrations in all the animals. Corticosterone level was increased in plasma of young but not of adult animals. Granulosa cells isolated from the ovaries of does subjected to high temperatures released less progesterone, and they had poorer response to the stimulatory action of FSH than the cells from control does. High temperatures induced fragmentation of nucleoli in ovarian granulosa cells, but they did not alter the state of other organelles in ovarian, testicular, or adrenocortical cells. A negative influence of high temperature on rabbit feed consumption, growth, viability, and fecundity was observed. Taken together, these changes could be due to a decrease in IGF-I and/or progesterone secretion, destruction of ovarian cell nucleoli, and/or impaired ovarian cell response to FSH.
Asunto(s)
Conducta Alimentaria/fisiología , Calor , Conejos/crecimiento & desarrollo , Conejos/fisiología , Sobrevida , Envejecimiento , Animales , Femenino , Hormona Folículo Estimulante/farmacología , Células de la Granulosa/efectos de los fármacos , Células de la Granulosa/fisiología , Masculino , Progesterona/metabolismoRESUMEN
The existing knowledge of the direct action of kisspeptin on the ovary needs to be expanded. In our study, the direct effects of kisspeptin on basic ovarian cell functions and their response to FSH were examined. We studied the effect of kisspeptin alone (0, 1, 10, and 100 ng/mL) and of kisspeptin (1, 10, and 100 ng/mL) in combination with FSH (10 ng/mL) on cultured porcine granulosa cells. Markers of viability, proliferation (accumulation of proliferating cell nuclear antigen [PCNA] and cyclin B1), and apoptosis (accumulation of bax and caspase 3), as well as the release of steroid hormones and IGF-I were analyzed using the trypan blue exclusion test, quantitative immunocytochemistry, and ELISA. Addition of kisspeptin at lower doses (1 and 10 ng/mL) increased cell viability, the accumulation of PCNA and cyclin B1, decreased the accumulation of bax and caspase 3, and promoted release of progesterone, estradiol, and IGF-I, but not testosterone. A high dose (100 ng/mL) of kisspeptin had the opposite, inhibitory effect. The addition of FSH increased cell viability, proliferation, decreased apoptosis, and promoted progesterone, testosterone, estradiol, and IGF-I release. Kisspeptin at lower doses supported the stimulatory action of FSH on viability, PCNA and cyclin B1 accumulation, and release of progesterone and estradiol, promoted its inhibitory action on bax and caspase 3 accumulation, but did not modify its action on testosterone and IGF-I release. On the contrary, kisspeptin at a high dose inhibited and even reversed the FSH effect. FSH mimicked and promoted both the stimulatory and inhibitory action of kisspeptin on all examined ovarian functions besides IGF-I release. These observations show that kisspeptin can directly regulate basal ovarian cell functions. Furthermore, they demonstrate the functional interrelationships between kisspeptin and FSH in direct regulation of ovarian functions.
Asunto(s)
Hormona Folículo Estimulante/administración & dosificación , Células de la Granulosa/fisiología , Kisspeptinas/administración & dosificación , Ovario/fisiología , Sus scrofa/fisiología , Animales , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Hormonas Esteroides Gonadales/metabolismo , Células de la Granulosa/efectos de los fármacos , Ovario/efectos de los fármacosRESUMEN
The involvement of the mTOR system/enzyme sirtuin 1 (SIRT1) intracellular signaling system in the control of ovarian functions and its role in mediating hormonal action on the ovary has been proposed, but this hypothesis should be supported by a demonstrated influence of hormones on mTOR/SIRT1. Therefore, the aim of our in vitro experiments was to examine the effect of the known hormonal regulators of ovarian functions, such as follicle-stimulating hormone (FSH), oxytocin (OT) and insulin-like growth factor I (IGF-I), on mTOR/SIRT1. The accumulation of SIRT1 in porcine ovarian granulosa cells cultured with and without these hormones (at doses of 1, 10 or 100 ng.ml-1) was evaluated using immunocytochemistry. It was observed that the addition of FSH (at 10 ng.ml-1 but not at 1 or 100 ng/ml) and OT (at all tested doses) increased the expression of SIRT1 in ovarian cells. In addition, 100 ng.ml-1, but not at 1 or 10 ng.ml-1, of IGF-I decreased SIRT1 accumulation. Our observations are the first demonstration that hormones can directly regulate the ovarian mTOR/SIRT1 system and that this system could mediate the action of hormonal regulators on the ovary.
Asunto(s)
Hormona Folículo Estimulante/farmacología , Células de la Granulosa/metabolismo , Factor I del Crecimiento Similar a la Insulina/farmacología , Ovario/metabolismo , Oxitocina/farmacología , Sirtuina 1/biosíntesis , Animales , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Femenino , Células de la Granulosa/citología , Células de la Granulosa/efectos de los fármacos , Ovario/citología , Ovario/efectos de los fármacos , Oxitócicos/farmacología , Sirtuina 1/genética , Sirtuina 1/metabolismo , PorcinosRESUMEN
The influence of environmental contaminant toluene and of plant fennel (Foeniculum vulgare Mill.) on reproduction are reported, but the mechanisms of their action and the protective effect of fennel on contaminant influence remain to be elucidated. In this study, we hypothesized that toluene and fennel directly affects basic ovarian cell functions, and that fennel can be used as an appropriate natural protective agent against the potential adverse effects of toluene. This study aimed to examine the action of toluene (20 µg/mL) and fennel extract (0, 1, 10, 100 µg/mL), and assess their combination on viability, proliferation, apoptosis, and hormone release by cultured healthy mare ovarian granulosa cells. Viability, proliferation (percentage of PCNA-positive cells), apoptosis and release of progesterone, oxytocin and prostaglandin F were evaluated by using Trypan blue exclusion tests, immunocytochemistry and enzyme immunoassays, respectively. Toluene, when given alone, inhibited viability, proliferation, apoptosis, progesterone, prostaglandin F and IGF-I. However, it did not affect oxytocin release. Moreover, Fennel, when given alone, inhibited viability, progesterone, and prostaglandin F release, as well as stimulating proliferation and oxytocin release. In addition, Fennel did not affect apoptosis. When given in combination with toluene, fennel was able to suppress, and even invert, the effects of toluene on viability, proliferation, apoptosis, prostaglandin F, and IGF-I. However, it did not alter its effect on progesterone release. Moreover, fennel induced the inhibitory effect of toluene on oxytocin output. The findings of our study suggest direct adverse effects of toluene on the basic ovarian functions of mares. Lastly, we also observed the direct influence of fennel on these functions, as well as its ability to be a natural protector against the action of toluene on the ovarian functions of mares.
Asunto(s)
Foeniculum/química , Células de la Granulosa/efectos de los fármacos , Tolueno/toxicidad , Animales , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Femenino , Caballos , Factor I del Crecimiento Similar a la Insulina/metabolismo , Oxitocina/farmacología , Extractos Vegetales/farmacología , Progesterona/farmacología , Prostaglandinas F/metabolismoRESUMEN
The present in vitro study was conducted to examine the direct action of the plant steroidal sapogenin, diosgenin, on basic farm animal ovarian cell functions. As models, we used cultured porcine ovarian granulosa cells, porcine whole follicles, and rabbit ovarian fragments. The effects of diosgenin (0, 1, 10, or 100 µg/mL medium) on the markers of proliferation, cytoplasmic apoptosis, steroid (progesterone: P4, testosterone: T, and estradiol: E2) release, and peptide hormone (insulin-like growth factor I: IGF-I) release were analyzed by quantitative immunocytochemistry and radioimmunoassay. Diosgenin promoted proliferation, apoptosis, and T and E2 release and inhibited P4 output in cultured porcine granulosa cells. Similarly, cultured porcine ovarian follicles showed diosgenin-induced inhibition of P4 and stimulation of T release. In cultured rabbit ovarian fragments, diosgenin stimulated P4 and IGF-I release. This is the first study showing that diosgenin can promote basic ovarian cell functions such as proliferation, apoptosis, and steroid and peptide hormone release. The similar effects of diosgenin on porcine granulosa cells and ovarian follicles suggest that granulosa cells are the primary ovarian target of diosgenin. The contrasting effects of diosgenin on porcine and rabbit ovarian P4 output suggest that diosgenin functions in a species-specific manner. These observations indicate that diosgenin has potential applications for improving female reproduction.
Asunto(s)
Diosgenina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Ovario/efectos de los fármacos , Conejos , Porcinos , Animales , Femenino , Inmunohistoquímica/veterinaria , Factor I del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Ovario/metabolismo , Progesterona/genética , Progesterona/metabolismoRESUMEN
The aim of the present study was to examine the role of cAMP response element-binding protein (CREB) and its phosphorylation in the regulation of ovarian cell proliferation and apoptosis, and of the response of proliferation and apoptosis to the upstream hormonal stimulators FSH and insulin-like growth factor (IGF) 1. In the first series of experiments, porcine ovarian granulosa cells, transfected or not with a gene construct encoding wild-type CREB1 (CREB1WT), were cultured with and without FSH (0, 1, 10 or 100ngmL-1). In the second series of experiments, these cells were transfected or not with CREB1WT or non-phosphorylatable mutant CREB1 (CREB1M1) and cultured with and without FSH (0, 1, 10 or 100ngmL-1) or IGF1 (0, 1, 10 and 100ngmL-1). Levels of total and phosphorylated (p-) CREB1, proliferating cell nuclear antigen (PCNA), a marker of proliferation, and BAX, a marker of apoptosis, were evaluated by western immunoblotting and immunocytochemical analysis. Transfection of cells with CREB1WT promoted accumulation of total CREB1 within cells, but p-CREB1 was not detected in any cell group. Both CREB1WT and CREB1M1 reduced cell proliferation and apoptosis. Addition of 10 and 100ngmL-1 FSH to non-transfected cells promoted CREB1 accumulation and apoptosis, whereas cell proliferation was promoted by all concentrations of FSH tested. FSH activity was not modified in cells transfected with either CREB1WT or CREB1M1. IGF1 at 100ngmL-1 promoted cell proliferation, whereas all concentrations of IGF1 tested reduced apoptosis. Transfection with either CREB1WT or CREB1M1 did not modify the effects of either FSH or IGF1, although CREB1M1 reversed the effect of IGF1 on apoptosis from inhibitory to stimulatory. These observations suggest that CREB1 is involved in the downregulation of porcine ovarian cell proliferation and apoptosis. The absence of visible CREB1 phosphorylation and the similarity between the effects of CREB1WT and CREB1M1 transfection indicate that phosphorylation is not necessary for CREB1 action on these processes. Furthermore, the observations suggest that FSH promotes both ovarian cell proliferation and apoptosis, whereas IGF1 has proliferation-promoting and antiapoptotic properties. The effect of FSH on CREB1 accumulation and the ability of CREB1M1 to reverse the effects of IGF1 on apoptosis indicate that CREB1 is a mediator of hormonal activity, but the inability of either CREB1WT or CREBM1transfection to modify the primary effects of FSH and IGF1 suggest that CREB1 and its phosphorylation do not mediate the action of these hormones on ovarian cell proliferation and apoptosis.
Asunto(s)
Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Hormona Folículo Estimulante/farmacología , Factor I del Crecimiento Similar a la Insulina/farmacología , Ovario/efectos de los fármacos , Animales , Apoptosis/fisiología , Proliferación Celular/fisiología , Femenino , Ovario/metabolismo , Fosforilación/efectos de los fármacos , PorcinosRESUMEN
The body condition score (BCS) of cows affects their reproductive efficiency, but the underlying mechanism is unclear. We examined the effect of BCS on the basic ovarian cell functions and their responses to gonadotropic and metabolic hormones. We isolated ovarian cells from cows with a tendency toward emaciation (BCS2) and those with an average body condition (BCS3), and we compared their hormonal release and responses to FSH, leptin, ghrelin, and neuropeptide Y (NPY) added at doses of 0, 1, 10, or 100â¯ng/mL. Progesterone, testosterone, estradiol, and insulin-like growth factor I (IGF-I) release were evaluated by RIA. No differences were found in progesterone or testosterone release between BCS2 and BCS3 cells; however, ovarian cells from BCS2 cows released more estradiol and IGF-I than cells from BCS3 cows. FSH, ghrelin, and NPY promoted progesterone release in BCS2 cells but had no stimulatory or inhibitory effect on BCS3 cells. In contrast, leptin promoted progesterone release in BCS3 cells and inhibited progesterone release in BCS2 cells. FSH also promoted testosterone release in both BCS2 and BCS3 cells but inhibited progesterone at a low dose in BCS3 cells. Leptin inhibited testosterone release in BCS3 cells but not in BCS2 cells. Estradiol release was promoted by leptin and ghrelin in BCS3 cells; however, it was unaffected by leptin and inhibited by ghrelin in BCS2 cells. IGF-I production was promoted by FSH and inhibited by leptin in both groups. Ghrelin suppressed IGF-I release in BCS2 cells and increased IGF-I release in BCS3 cells. NPY promoted IGF-I release in BCS2 cells but not in BCS3 cells. Our results demonstrate the effects of BCS on ovarian cell estradiol and IGF-I (but not progesterone or testosterone) release, as well as on the responses of ovarian cells to FSH, leptin, ghrelin, and NPY.
Asunto(s)
Constitución Corporal/fisiología , Bovinos , Ghrelina/farmacología , Hormonas Esteroides Gonadales/farmacología , Leptina/farmacología , Ovario/efectos de los fármacos , Ovario/metabolismo , Animales , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Estradiol/farmacología , Femenino , Células de la Granulosa/efectos de los fármacos , Células de la Granulosa/metabolismo , Factor I del Crecimiento Similar a la Insulina/farmacología , Ovario/citología , Cultivo Primario de Células , Progesterona/farmacología , Testosterona/farmacologíaRESUMEN
The polyphenol-rich plants rooibos and ginkgo are widely used in folk medicine and in preparation of foods and drinks, but their effect on reproduction has not been properly studied yet. The aim of our in vitro experiments was to examine the possible direct effect of rooibos and ginkgo on the basic ovarian cell functions-proliferation, apoptosis and release of hormones progesterone (P4) and leptin (L). Porcine ovarian granulosa cells were cultured in the presence of rooibos or ginkgo extract (0, 1, 10 and 100 µg/ml of medium). The accumulation of markers of proliferation (PCNA and cyclin B1) and apoptosis (bax) and their mRNAs was analysed using immunocytochemistry and by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Release of P4 and L was evaluated by radioimmunoassay. It was observed that rooibos or ginkgo addition was able to inhibit proliferation (down-regulates PCNA, cyclin B1 and their mRNAs), to promote apoptosis (accumulation of bax) and to suppress both L and P4 release by ovarian cells. These data suggest a direct inhibitory (anti-proliferative, pro-apoptotic and hormone-suppressing) effect of polyphenol-containing plants rooibos and ginkgo on ovarian functions. The potential anti-reproductive effect of these medical plants is to be taken into account by their consumption.
Asunto(s)
Aspalathus/química , Ginkgo biloba/química , Células de la Granulosa/efectos de los fármacos , Extractos Vegetales/farmacología , Polifenoles/química , Porcinos/fisiología , Animales , Biomarcadores , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Células de la Granulosa/fisiología , Extractos Vegetales/administración & dosificación , Extractos Vegetales/químicaRESUMEN
The present study investigated whether dietary turmeric (Curcuma longa L.) can improve rabbit reproduction, ovarian function, growth, or viability. Female New Zealand White rabbits were either fed a standard diet (n=15) or a diet enriched with 5 g (group E1) or 20 g (group E2) turmeric powder per 100 kg feed mixture (n=16 or 15, respectively). After 295 days, weight gain, conception and kindling rates, pup and mother viability, ovarian macro- and micro-morphometric indices, release of leptin in response to the addition LH, and the release of progesterone, testosterone and leptin by isolated ovarian fragments were analyzed. Dietary turmeric failed to affect ovarian length and weight but did increase the number of primary follicles (E2: 32.5% greater than control group), as well as the diameter of primary (E1: +19.4%, E2: +21.1%), secondary (E2: +41.4%), and tertiary (E1: +97.1%, E2: +205.1%) follicles. Turmeric also increased the number of liveborn (E1: +21.0%) and weaned (E1: +25.0%) pups and decreased the number of stillborn pups (E2: -87.5%) but did not affect weight gain, conception, or kindling rate. Furthermore, dietary turmeric decreased doe mortality during the first reproductive cycle (13.3% in control; 0% in E1; and 6.7% in E2) but not during the second cycle. In vitro, the ovaries of the turmeric-treated rabbits released more progesterone (E1: +85.7%, E2: +90.0%) and less testosterone (E2: -87.0%) and leptin (E2: -29.0%) than the ovaries of control rabbits. Moreover, LH decreased the leptin output of control rabbits but increased that of experimental rabbits. Therefore, it is likely that dietary turmeric improves pup viability and that it could promote rabbit fecundity by either (1) promoting the production of primary ovarian follicles or (2) stimulating the growth of follicles at all stages of folliculogenesis.
Asunto(s)
Curcuma , Fertilidad , Folículo Ovárico , Conejos , Animales , Curcuma/química , Estradiol , Femenino , Fertilidad/efectos de los fármacos , Hormona Folículo Estimulante , Hormona Luteinizante/fisiología , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/crecimiento & desarrollo , Ovario , Progesterona , Conejos/fisiologíaRESUMEN
T-2 toxin and its metabolite HT-2 toxin are one of the most toxic mycotoxins of type A-trichothecenes, which are produced mainly by Fusarium species. Therefore, study of Fusarium toxins T-2 toxin and HT-2 toxin is an essential issue because they could also play role in failures of reproductive functions as well as endocrine system of domestic animals. Assessment of the effect of A-trichothecene mycotoxin HT-2 toxin alone or combined with insulin-like growth factor (IGF-I), leptin and ghrelin on estradiol secretion by rabbit ovarian fragments in vitro was done. Rabbit ovarian fragments were incubated without (control group) or with HT-2 toxin, or its combinations with IGF-I, leptin and ghrelin at various concentrations for 24 h. Secretion of 17beta-estradiol was determined by ELISA. Firstly, HT-2 toxin at the doses 10 and 100 ng.ml(-1), but not at 1 ng.ml(-1) decreased 17beta-estradiol secretion by ovarian fragments. Secondly, 17beta-estradiol secretion was not affected by HT-2 toxin exposure combined with growth factor IGF-I, metabolic hormones leptin and ghrelin. In conclusion, HT-2 toxin has potent direct dose-dependent effects on ovarian steroidogenesis in rabbits. These direct effects of HT-2 mycotoxin on ovarian steroidogenesis could impact negatively on the reproductive performance of rabbits.