RESUMEN
BACKGROUND: Nicotine, a pharmacologically active component of tobacco adversely affects the male reproductive system and fertility whereas icariin (ICA), the main active ingredient in Epimedium herba has been used in the treatment of several male reproductive problems. This study aimed at evaluating the protective or ameliorative effect of ICA against reproductive toxicity induced by intraperitoneal injection of nicotine in mice. METHODS: Using simple random allocation, forty male mice were randomly divided into 4 groups: control (received 0.35 mL physiological saline via gastric gavage), nicotine (0.75 mg/kg BW/day intraperitoneally), ICA (75 mg/kg BW/day gastric gavage), and nicotine plus ICA (nicotine, 0.75 mg/kg BW/day intraperitoneally + ICA, 75 mg/kg BW/day gastric gavage) group. After 35 days of treatment, the mice were weighed, sacrificed, and their reproductive organs (testis and epididymis) were collected and examined for further studies. RESULTS: The nicotine-treated group showed significantly decreased epididymal sperm density and serum testosterone concentration relative to the control group. Nicotine also caused oxidative damage shown by significant reduction in the activities of antioxidant enzymes and elevation in Malondialdehyde (MDA) levels. ICA on the other hand, improved the reduction in sperm density, hormone levels, and activities of antioxidant enzymes altered in the nicotine treated mice. CONCLUSIONS: These findings indicate that nicotine-induced reproductive toxicity and oxidative damage on male reproductive tissues could be attenuated by ICA.
Asunto(s)
Epidídimo/efectos de los fármacos , Flavonoides/farmacología , Nicotina/toxicidad , Agonistas Nicotínicos/toxicidad , Sustancias Protectoras/farmacología , Espermatozoides/efectos de los fármacos , Testículo/efectos de los fármacos , Testosterona/sangre , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/genética , Animales , Peso Corporal/efectos de los fármacos , Epidídimo/patología , Glutatión Peroxidasa/efectos de los fármacos , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/metabolismo , Masculino , Malondialdehído/metabolismo , Ratones , Tamaño de los Órganos/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Fosfoproteínas/efectos de los fármacos , Fosfoproteínas/genética , Distribución Aleatoria , Recuento de Espermatozoides , Espermatozoides/patología , Superóxido Dismutasa/efectos de los fármacos , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Testículo/metabolismo , Testículo/patologíaRESUMEN
At present, there is no reliable in vitro assembled prepubertal testis-like biomimetic organ culture system designed to assess the functional effects of human gonadotropins on Sertoli and Leydig cells. Spermatogenesis is regulated by endocrine, paracrine, and juxtacrine factors (testicular cross-talk), mainly orchestrated by gonadotropins such as luteinizing hormone (LH) and follicle-stimulating hormone (FSH) that play a pivotal role by stimulating Leydig and Sertoli cells, respectively. The aim of our study was to set up an in vitro prepubertal porcine bioengineered construct as a new model for experimental studies on reassembled Sertoli and Leydig cells. We have evaluated Sertoli and Leydig cells obtained from 15- to 20-day-old neonatal pig testes in terms of purity and function. Subsequently, purified Sertoli and enriched Leydig cells were subjected to coincubation to obtain an in vitro prepubertal porcine testis-like culture system. We performed enzyme-linked immunosorbent assay (ELISA) for anti-Müllerian hormone (AMH), inhibin B, and testosterone secretion in the medium, and Real-Time PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3ß-HSD mRNA expression levels. This in vitro testis-like system was highly responsive to the effects of human gonadotropins and testosterone. AMH mRNA expression and secretion declined, and inhibin-B increased, while FSH-receptor expression was downregulated upon FSH/LH exposure/treatment. Finally, the production of testosterone was increased selectively upon LH treatment. In summary, our proposed model could help to better determine the action of human gonadotropins on Sertoli and Leydig cells. The potential usefulness of the system for shedding light into male infertility-related issues is evident.
Asunto(s)
Hormona Folículo Estimulante/farmacología , Hormonas/farmacología , Células Intersticiales del Testículo/efectos de los fármacos , Hormona Luteinizante/farmacología , Células de Sertoli/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/genética , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Animales Recién Nacidos , Hormona Antimülleriana/genética , Hormona Antimülleriana/metabolismo , Aromatasa/efectos de los fármacos , Aromatasa/genética , Aromatasa/metabolismo , Técnicas de Cultivo de Célula , Ensayo de Inmunoadsorción Enzimática , Técnicas In Vitro , Inhibinas/efectos de los fármacos , Inhibinas/genética , Inhibinas/metabolismo , Células Intersticiales del Testículo/metabolismo , Masculino , Modelos Biológicos , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de HFE/efectos de los fármacos , Receptores de HFE/genética , Receptores de HFE/metabolismo , Receptores de HL/efectos de los fármacos , Receptores de HL/genética , Receptores de HL/metabolismo , Células de Sertoli/metabolismo , Porcinos , Testículo/efectos de los fármacos , Testículo/metabolismo , Testosterona/metabolismoRESUMEN
This study was to investigate the effect of annexin A5 on testosterone secretion from primary rat Leydig cells and the underlying mechanisms. Isolated rat Leydig cells were treated with annexin A5. Testosterone production was detected by chemiluminescence assay. The protein and mRNA of Steroidogenic acute regulatory (StAR), P450scc, 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase (17ß-HSD), and 17α-hydroxylase were examined by Western blotting and semi-quantitative RT-PCR, respectively. Annexin A5 significantly stimulated testosterone secretion from rat Leydig cells in dose- and time-dependent manners and increased mRNA and protein expression of StAR, P450scc, 3ß-HSD, and 17ß-HSD but not 17α-hydroxylase. Annexin A5 knockdown by siRNA significantly decreased the level of testosterone and protein expression of P450scc, 3ß-HSD, and 17ß-HSD. The significant activation of ERK1/2 signaling was observed at 5, 10, and 30 min after annexin A5 treatment. After the pretreatment of Leydig cells with ERK inhibitor PD98059 (50 µmol l-1 ) for 20 min, the effects of annexin A5 on promoting testosterone secretion and increasing the expression of P450scc, 3ß-HSD, and 17ß-HSD were completely abrogated (P < 0.05). Thus, ERK1/2 signaling is involved in the roles of annexin A5 in mediating testosterone production and the expression of P450scc, 3ß-HSD, and 17ß-HSD in Leydig cells.
Asunto(s)
Anexina A5/farmacología , Inhibidores Enzimáticos/farmacología , Células Intersticiales del Testículo/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , ARN Mensajero/efectos de los fármacos , Testosterona/biosíntesis , 17-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/genética , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/genética , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Western Blotting , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/efectos de los fármacos , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/metabolismo , Células Intersticiales del Testículo/metabolismo , Masculino , Fosfoproteínas/efectos de los fármacos , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , ARN Mensajero/metabolismo , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Growth differentiation factor-8 (GDF-8) has been recently shown to be expressed in human granulosa cells, and the mature form of GDF-8 protein can be detected in the follicular fluid. However, the biological function and significance of this growth factor in the human ovary remains to be determined. Here, we investigated the effects of GDF-8 on steroidogenic enzyme expression and the potential mechanisms of action in luteinized human granulosa cells. We demonstrated that treatment with GDF-8 did not affect the mRNA levels of P450 side-chain cleavage enzyme and 3ß-hydroxysteroid dehydrogenase, whereas it significantly down-regulated steroidogenic acute regulatory protein (StAR) expression and decreased progesterone production. The suppressive effect of GDF-8 on StAR expression was abolished by the inhibition of the TGF-ß type I receptor. In addition, treatment with GDF-8 activated both Smad2/3 and ERK1/2 signaling pathways. Furthermore, knockdown of activin receptor-like kinase 5 reversed the effects of GDF-8 on Smad2/3 phosphorylation and StAR expression. The inhibition of Smad3 or ERK1/2 signaling pathways attenuated the GDF-8-induced down-regulation of StAR and production of progesterone. Interestingly, the concentrations of GDF-8 were negatively correlated with those of progesterone in human follicular fluid. These results indicate a novel autocrine function of GDF-8 to down-regulate StAR expression and decrease progesterone production in luteinized human granulosa cells, most likely through activin receptor-like kinase 5-mediated Smad3 and ERK1/2 signaling pathways. Our findings suggest that granulosa cells might play a critical role in the regulation of progesterone production to prevent premature luteinization during the final stage of folliculogenesis.
Asunto(s)
Células de la Granulosa/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Miostatina/farmacología , Proteínas Serina-Treonina Quinasas/efectos de los fármacos , ARN Mensajero/efectos de los fármacos , Receptores de Factores de Crecimiento Transformadores beta/efectos de los fármacos , Proteína smad3/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/genética , Línea Celular , Regulación hacia Abajo , Femenino , Líquido Folicular/química , Células de la Granulosa/metabolismo , Humanos , Luteinización , Fosfoproteínas/efectos de los fármacos , Fosfoproteínas/genética , Fosforilación/efectos de los fármacos , Progesterona/biosíntesis , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Mensajero/metabolismo , Receptor Tipo I de Factor de Crecimiento Transformador beta , Receptores de Factores de Crecimiento Transformadores beta/antagonistas & inhibidores , Receptores de Factores de Crecimiento Transformadores beta/metabolismo , Transducción de Señal , Proteína Smad2/efectos de los fármacos , Proteína Smad2/metabolismo , Proteína smad3/metabolismoRESUMEN
Urocortin 2 (UCN2) is a neuropeptide of the CRH family, involved in homeostatic mechanisms, the stress response, and control of anxiety. To elucidate the effects of UCN2 on steroidogenesis, we developed a mouse model that allows a Cre recombinase-determined conditional overexpression of UCN2 (UCN2-COE). In these mice SF1-Cre-driven overexpression of UCN2 was restricted to the adrenal glands, gonads, and parts of the hypothalamus. UCN2-COE animals of both sexes revealed significantly higher plasma UCN2 levels and significantly higher UCN2 expression levels in the adrenals and ovaries. In contrast, the baseline expression of UCN2 was already high in the testes of control mice with no further increase achievable in UCN2-COE animals. Adrenal steroidogenesis of UCN2-COE animals was investigated under baseline conditions, upon an ACTH stimulation test, and following a restraint stress test. A tendency toward lower expression of steroidogenic enzymes was detectable in UCN2-COE animals of both sexes with slight differences between males and females. A similar reduction in the expression levels of the final steps of ovarian steroidogenesis, accompanied by reduced plasma estradiol levels, was observed in female UCN2-COE animals. Thus, adrenal UCN2 overexpression resulted in down-regulation of adrenal steroidogenesis, suggesting a reduction in the stress response in the mouse (stress coping behavior). Similarly, UCN2 overexpression in the ovaries caused a decrease in steroidogenesis and reduction of follicles that had undergone ovulation. Nevertheless, this finding was not associated with reduced fertility.
Asunto(s)
Glándulas Suprarrenales/metabolismo , Hormona Liberadora de Corticotropina/genética , Ovario/metabolismo , ARN Mensajero/metabolismo , Urocortinas/genética , 17-Hidroxiesteroide Deshidrogenasas/genética , 17-alfa-Hidroxipregnenolona/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/genética , Glándulas Suprarrenales/efectos de los fármacos , Hormona Adrenocorticotrópica/farmacología , Animales , Aromatasa/metabolismo , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/efectos de los fármacos , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Citocromo P-450 CYP11B2/efectos de los fármacos , Citocromo P-450 CYP11B2/genética , Estradiol/metabolismo , Femenino , Técnicas de Sustitución del Gen , Hormonas Esteroides Gonadales , Masculino , Ratones , Ovario/anatomía & histología , Fenotipo , Fosfoproteínas/efectos de los fármacos , Fosfoproteínas/genética , Progesterona/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Esteroide 11-beta-Hidroxilasa/efectos de los fármacos , Esteroide 11-beta-Hidroxilasa/genética , Esteroide 17-alfa-Hidroxilasa/genética , Testículo/metabolismoRESUMEN
The objective of the present study was to investigate the effects of testosterone in recuperation of lead-induced suppressed reproduction in adult male rats. Lead acetate was administered orally to adult male rats (95 ± 5 days) at dosage level of 0.05 and 0.15% for 55 days through drinking water and injected intraperitoneally with either testoviron depot at a dose of 4.16 mg kg(-1) body weight or vehicle alone on days 1, 7 and 14 respectively. At the end of treatment, control and treated males were cohabited with untreated normal-cycling females. After cohabitation for 5 days, all the male rats were killed and weights of reproductive organs were determined. Significant increase in the indices of testis, epididymis, seminal vesicles, vas deferens and prostate glands was observed in testosterone (T)-treated rats when compared to those of lead-exposed rats. Testosterone treatment significantly increased epididymal sperm count, motile spermatozoa, viable spermatozoa and HOS tail-coiled spermatozoa and also the activity levels of testicular 3ß- and 17ß-hydroxysteroid dehydrogenases when compared to those of lead-exposed males. From the results, it can be hypothesised that supplementation of testosterone mitigated lead-induced suppressed reproduction in male rats.
Asunto(s)
Andrógenos/farmacología , Infertilidad Masculina/inducido químicamente , Compuestos Organometálicos/toxicidad , Espermatozoides/efectos de los fármacos , Testículo/efectos de los fármacos , Testosterona/farmacología , 17-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Epidídimo/efectos de los fármacos , Epidídimo/patología , Hormona Folículo Estimulante/sangre , Hormona Luteinizante/sangre , Hormona Luteinizante/efectos de los fármacos , Masculino , Tamaño de los Órganos , Próstata/efectos de los fármacos , Próstata/patología , Ratas , Ratas Wistar , Vesículas Seminales/efectos de los fármacos , Vesículas Seminales/patología , Motilidad Espermática/efectos de los fármacos , Testículo/metabolismo , Testículo/patología , Conducto Deferente/efectos de los fármacos , Conducto Deferente/patologíaRESUMEN
Aberrant androgen receptor (AR) activation is the major driver of castrate resistant prostate cancer (CRPC). CRPC is ultimately fatal and more therapeutic agents are needed to treat this disease. Compounds that target the androgen axis by inhibiting androgen biosynthesis and or AR signaling are potential candidates for use in CRPC treatment and are currently being pursued aggressively. Aldo-keto reductase 1C3 (AKR1C3) plays a pivotal role in androgen biosynthesis within the prostate. It catalyzes the 17-ketoreduction of weak androgen precursors to give testosterone and 5α-dihydrotestosterone. AKR1C3 expression and activity has been implicated in the development of CRPC, making it a rational target. Selective inhibition of AKR1C3 will be important, however, due to the presence of closely related isoforms, AKR1C1 and AKR1C2 that are also involved in androgen inactivation. We examine the evidence that supports the vital role of AKR1C3 in CRPC and recent developments in the discovery of potent and selective AKR1C3 inhibitors. This article is part of a Special Issue entitled 'CSR 2013'.
Asunto(s)
3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , Hidroxiprostaglandina Deshidrogenasas/efectos de los fármacos , Neoplasias de la Próstata/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Miembro C3 de la Familia 1 de las Aldo-Ceto Reductasas , Humanos , Hidroxiprostaglandina Deshidrogenasas/metabolismo , Masculino , OrquiectomíaRESUMEN
The objective of the present study was to investigate the effects of genistein and equol on 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and 17beta-hydroxysteroid dehydrogenase 3 (17beta-HSD3) in human and rat testis microsomes. These enzymes (3beta-HSD and 17beta-HSD3), along with two others (cytochrome P450 side-chain cleavage enzyme and cytochrome P450 17alpha-hydroxylase/17-20 lyase), catalyze the reactions that convert the steroid cholesterol into the sex hormone testosterone. Genistein inhibited 3beta-HSD activity (0.2 micromol L(-1) pregnenolone) with half-maximal inhibition or a half-maximal inhibitory concentration (IC(50)) of 87 +/- 15 (human) and 636 +/- 155 nmol L(-1) (rat). Genistein's mode of action on 3beta-HSD activity was competitive for the substrate pregnenolonrge and noncompetitive for the cofactor NAD(+). There was no difference in genistein's potency of 3beta-HSD inhibition between intact rat Leydig cells and testis microsomes. In contrast to its potent inhibition of 3beta-HSD, genistein had lesser effects on human and rat 17beta-HSD3 (0.1 micromol L(-1) androstenedione), with an IC(50) >or= 100 micromol L(-1). On the other hand, equol only inhibited human 3beta-HSD by 42%, and had no effect on 3beta-HSD and 17beta-HSD3 in rat tissues. These observations imply that the ability of soy isoflavones to regulate androgen biosynthesis in Leydig cells is due in part to action on Leydig cell 3beta-HSD activity. Given the increasing intake of soy-based food products and their potential effect on blood androgen levels, these findings are greatly relevant to public health.
Asunto(s)
17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Genisteína/farmacología , Isoflavonas/farmacología , Testículo/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , Animales , Equol , Humanos , Cinética , Células Intersticiales del Testículo/enzimología , Masculino , Microsomas/efectos de los fármacos , Microsomas/enzimología , Ratas , Ratas Sprague-Dawley , Testículo/enzimologíaRESUMEN
OBJECTIVE: To compare the antifertility effects of 50% ethanolic extract of the root bark of Cananga odorata with gossypol. DESIGN: Controlled research laboratory study. SETTING: University research laboratory. ANIMAL(S): Male albino rats (Sprague Dawley, body weight 150 +/- 5 g) bred in university animal house. INTERVENTION(S): A 50% ethanolic extract of the root bark of Cananga odorata and gossypol was administered orally for 60 days. On day 61, one third of the animals in each group were killed for various analyses. One half of the remaining animals were used for the evaluation of fertility index, and other half was maintained for 15 days more on normal diet. MAIN OUTCOME MEASURE(S): The epididymal sperm motility, morphology, and count, the testicular activities of HMG CoA reductase, 3beta-hydroxy steroid dehydrogenase, glucose-6-phosphate dehydrogenase, and testicular cholesterol and serum testosterone were assessed. RESULT(S): Differences were not observed in the sperm count and fertility index of the gossypol group in comparison with the C odorata group. But statistically significant alterations were noted in the sperm morphology as well as in the activity of HMG CoA reductase, 3beta-hydroxy steroid dehydrogenase, glucose-6-phosphate dehydrogenase, cholesterol, and protein of the testis and in serum testosterone. On withdrawal of the drugs, sperms in C odorata group became completely motile but not in the gossypol group. The active component is a 52 kd protein. CONCLUSION(S): The ethanolic extract of Cananga odorata possesses antifertility effects.
Asunto(s)
Cananga , Anticonceptivos Masculinos/farmacología , Gosipol , Extractos Vegetales/farmacología , Espermicidas/farmacología , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Colesterol/metabolismo , Epidídimo/efectos de los fármacos , Epidídimo/fisiología , Etanol , Fertilidad/efectos de los fármacos , Fertilidad/fisiología , Hidroximetilglutaril-CoA Reductasas/efectos de los fármacos , Hidroximetilglutaril-CoA Reductasas/metabolismo , Masculino , Ratas , Ratas Sprague-Dawley , Recuento de Espermatozoides , Motilidad Espermática/efectos de los fármacos , Testículo/efectos de los fármacos , Testículo/enzimología , Testículo/metabolismo , Testosterona/sangreRESUMEN
We here present an optical method for monitoring the activity of the inducible aldo-keto reductases AKR1C2 and AKR1C3 in living human cells. The induction of these enzymes is regulated by the antioxidant response element (ARE), as demonstrated in recent literature, which in turn is dependent on the transcription factor Nrf2. The activation of ARE leads to the transcription of a coalition of cytoprotective enzymes and thus represents an important target for the development of new therapies in the area of neurodegenerative diseases and cancer. Through the use of Coumberone, a metabolic fluorogenic probe, and isoform-selective inhibitors, the upregulation of cellular stress markers AKR1C2 and AKR1C3 can be quantitatively measured in the presence of ARE activator compounds, via either a fluorimetric assay or fluorescence microscopy imaging of intact cells. The method has both high sensitivity and broad dynamic range, as demonstrated by induction studies in three cell lines with dramatically different metabolic capabilities (transfected monkey kidney COS-1 cells, human neuroblastoma IMR-32 cells, and human liver HepG2 cells). We applied the new method to examine a number of neurotrophic natural products (spirotenuipesine A, spirotenuipesine B, scabronine G-methylester, and panaxytriol), and discovered that panaxytriol, an active component of red ginseng extracts, is a potent ARE inducer. The upregulation of AKR1C enzymes, induced by chemically homogeneous panaxytriol, was partially dependent on PKC and PI3K kinases as demonstrated by the application of selective inhibitors. This cellular mechanism may account for panaxytriol's neurotrophic, neuroprotective, and anticancer properties. The protective effects of ARE inducers against tumorgenesis and neurodegeneration fuel the growing interest in this area of research and the method described here will greatly enable these endeavors.
Asunto(s)
3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , Enediinos/farmacología , Alcoholes Grasos/farmacología , Colorantes Fluorescentes/farmacología , Compuestos Heterocíclicos de 4 o más Anillos/farmacología , Hidroxiprostaglandina Deshidrogenasas/efectos de los fármacos , Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , Panax/química , 3-Hidroxiesteroide Deshidrogenasas/química , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Miembro C3 de la Familia 1 de las Aldo-Ceto Reductasas , Animales , Antineoplásicos/farmacología , Células COS , Línea Celular Tumoral , Chlorocebus aethiops , Citoprotección , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Enediinos/química , Activación Enzimática/efectos de los fármacos , Inducción Enzimática/efectos de los fármacos , Alcoholes Grasos/química , Colorantes Fluorescentes/química , Colorantes Fluorescentes/metabolismo , Compuestos Heterocíclicos de 4 o más Anillos/química , Compuestos Heterocíclicos de 4 o más Anillos/metabolismo , Humanos , Hidroxiprostaglandina Deshidrogenasas/química , Hidroxiprostaglandina Deshidrogenasas/metabolismo , Hidroxiesteroide Deshidrogenasas/química , Hidroxiesteroide Deshidrogenasas/metabolismo , Indometacina/farmacología , Microscopía Fluorescente/métodos , Modelos Biológicos , Estructura Molecular , Degeneración Nerviosa/prevención & control , Fármacos Neuroprotectores/farmacología , Sensibilidad y Especificidad , Regulación hacia Arriba/efectos de los fármacos , Ácido Ursodesoxicólico/farmacologíaRESUMEN
Methoxychlor, an organochlorine pesticide, has been reported to induce reproductive abnormalities in male reproductive tract. To get more insight into the mechanism(s) of gonadal toxicity provoked by methoxychlor, we investigated whether treatment with methoxychlor at low observed adverse effect level (LOAEL) would alter the activities of steroidogenic enzymes such as Delta(5)3beta-hydroxysteroid dehydrogenase (3beta-HSD) and Delta(5)17beta-hydroxysteroid dehydrogenase (17beta-HSD), the expression levels of steroidogenic acute regulatory (StAR) protein and androgen binding protein (ABP) in the testis of adult male rats. The experimental rats were exposed to a single dose of methoxychlor (50 mg/kg body weight) orally. The rats were killed at 0, 3, 6, 12, 24 and 72 h following treatment using anesthetic ether and testes were collected, processed and used to measure the activities of 3beta-HSD, 17beta-HSD, levels of hydrogen peroxide produced and the expression levels of StAR protein, and ABP. Methoxychlor administration resulted in a sequential reduction in the expression of StAR protein and activities of 3beta-HSD, 17beta-HSD with concomitant increase in the levels of hydrogen peroxide in the testis. These changes were significant between 6-12 h following treatment. The levels of ABP declined at 6-12 h following exposure to methoxychlor. The present study demonstrates transient effect of methoxychlor at LOAEL on testicular steroidogenesis and the possible role of hydrogen peroxide in mediating these effects.
Asunto(s)
Regulación de la Expresión Génica/efectos de los fármacos , Insecticidas/toxicidad , Metoxicloro/toxicidad , Testículo/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Administración Oral , Proteína de Unión a Andrógenos/efectos de los fármacos , Proteína de Unión a Andrógenos/metabolismo , Animales , Peróxido de Hidrógeno/metabolismo , Insecticidas/administración & dosificación , Masculino , Metoxicloro/administración & dosificación , Fosfoproteínas/efectos de los fármacos , Fosfoproteínas/metabolismo , Ratas , Ratas Wistar , Testículo/metabolismo , Factores de TiempoRESUMEN
Although it is known that glucocorticoids induce differentiation of growth hormone (GH)-producing cells in rodents and birds, the effect of mineralocorticoids on GH mRNA expression and the origin of corticosteroids affecting somatotrope differentiation have not been elucidated. In this study, we therefore carried out experiments to determine the effect of mineralocorticoids on GH mRNA expression in the chicken anterior pituitary gland in vitro and to determine whether corticosteroids are synthesized in the chicken embryonic pituitary gland. In a pituitary culture experiment with E11 embryos, both corticosterone and aldosterone stimulated GH mRNA expression and increased the number of GH cells in both lobes of the pituitary gland in a dose-dependent manner. These effects of the corticosteroids were significantly reversed by pretreatment with mifepristone, a glucocorticoid receptor (GR) antagonist, or spironolactone, a mineralocorticoid receptor (MR) antagonist. Interestingly, an in vitro serum-free culture experiment with an E11 pituitary gland showed that the GH mRNA level spontaneously increased during cultivation for 2 days without any extra stimulation, and this increase in GH mRNA level was completely suppressed by metyrapone, a corticosterone-producing enzyme P450C11 inhibitor. Moreover, progesterone, the corticosterone precursor, also stimulated GH mRNA expression in the cultured chicken pituitary gland, and this effect was blocked by pretreatment with metyrapone. We also detected mRNA expression of enzymes of cytochrome P450 cholesterol side chain cleavage (P450scc) and 3beta-hydroxysteroid dehydrogenase1 (3beta-HSD1) in the developmental chicken pituitary gland from E14 and E18, respectively. These results suggest that mineralocorticoids as well as glucocorticoids can stimulate GH mRNA expression and that corticosteroids generated in the embryonic pituitary gland by intrinsic steroidogenic enzymes stimulate somatotrope differentiation.
Asunto(s)
Corticoesteroides/farmacología , Diferenciación Celular/efectos de los fármacos , Hipófisis/citología , Hipófisis/efectos de los fármacos , Somatotrofos/citología , Somatotrofos/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/genética , Aldosterona/farmacología , Animales , Embrión de Pollo , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/efectos de los fármacos , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Hormona del Crecimiento/efectos de los fármacos , Hormona del Crecimiento/genética , Hibridación in Situ , Metirapona/farmacología , Técnicas de Cultivo de Órganos , Hipófisis/metabolismo , Progesterona/farmacología , ARN Mensajero/efectos de los fármacos , ARN Mensajero/genética , Receptores de Mineralocorticoides/efectos de los fármacos , Receptores de Mineralocorticoides/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The H295R cell bioassay was used to evaluate the potential endocrine disrupting effects of 18 of the most commonly used pharmaceuticals in the United States. Exposures for 48 h with single pharmaceuticals and binary mixtures were conducted; the expression of five steroidogenic genes, 3betaHSD2, CYP11beta1, CYP11beta2, CYP17 and CYP19, was quantified by Q-RT-PCR. Production of the steroid hormones estradiol (E2), testosterone (T) and progesterone (P) was also evaluated. Antibiotics were shown to modulate gene expression and hormone production. Amoxicillin up-regulated the expression of CYP11beta2 and CYP19 by more than 2-fold and induced estradiol production up to almost 3-fold. Erythromycin significantly increased CYP11beta2 expression and the production of P and E2 by 3.5- and 2.4-fold, respectively, while production of T was significantly decreased. The beta-blocker salbutamol caused the greatest induction of CYP17, more than 13-fold, and significantly decreased E2 production. The binary mixture of cyproterone and salbutamol significantly down-regulated expression of CYP19, while a mixture of ethynylestradiol and trenbolone, increased E2 production 3.7-fold. Estradiol production was significantly affected by changes in concentrations of trenbolone, cyproterone, and ethynylestradiol. Exposures with individual pharmaceuticals showed the possible secondary effects that drugs may exert on steroid production. Results from binary mixture exposures suggested the possible type of interactions that may occur between drugs and the joint effects product of such interactions. Dose-response results indicated that although two chemicals may share a common mechanism of action the concentration effects observed may be significantly different.
Asunto(s)
Carcinoma Corticosuprarrenal/metabolismo , Sistema Enzimático del Citocromo P-450/efectos de los fármacos , Disruptores Endocrinos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Hormonas Esteroides Gonadales/biosíntesis , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/genética , Aromatasa/efectos de los fármacos , Aromatasa/genética , Línea Celular Tumoral , Citocromo P-450 CYP11B2/efectos de los fármacos , Citocromo P-450 CYP11B2/genética , Sistema Enzimático del Citocromo P-450/genética , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Estradiol/biosíntesis , Humanos , Progesterona/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Esteroide 11-beta-Hidroxilasa/efectos de los fármacos , Esteroide 11-beta-Hidroxilasa/genética , Esteroide 17-alfa-Hidroxilasa/efectos de los fármacos , Esteroide 17-alfa-Hidroxilasa/genética , Testosterona/biosíntesisRESUMEN
Pregnant Wistar strain albino rats were administered hydroxyprogesterone and the steroidogenic potential of the testis was analysed in the next generation adult male rats. In utero exposure to supranormal levels (10 mg/kg body weight or 25 mg/kg body weight) of hydroxyprogesterone decreased the activity levels of marker steroidogenic enzymes (3beta-hydroxysteroid dehydrogenase and 17beta-hydroxysteroid dehydrogenase) of adult (mature) rat testis. Hydroxyprogesterone mediation in the suppression of rat testicular steroidogenesis is suggested, which can be correlated to the observed decrease in male reproductive potential in mammals exposed to female hormones during embryonic development.
Asunto(s)
17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Hidroxiprogesteronas/farmacología , Efectos Tardíos de la Exposición Prenatal , 17-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , Animales , Femenino , Cinética , Masculino , NAD/metabolismo , Embarazo , RatasRESUMEN
EDS alkylating agent has been shown to selectively and temporarily kill LCs in adult rats. The first newly formed single LCs appeared at 14th day post ESD and showed detectable activity for 3beta-HSD and NADH2-diaphorase, which became progressively stronger with time after treatment The ultrastructural study revealed that the progenitor LCs differentiated into immature LCs within a week, and two weeks later they were transformed into mature LCs. Therefore, the restoration of new LC population after EDS treatment repeated the dynamics of normal LC development within a similar time range. The dynamics of enzyme activity correlated with structural differentiation of the new LC population.
Asunto(s)
Diferenciación Celular/efectos de los fármacos , Células Intersticiales del Testículo/efectos de los fármacos , Células Intersticiales del Testículo/enzimología , Mesilatos/farmacología , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Dihidrolipoamida Deshidrogenasa/efectos de los fármacos , Dihidrolipoamida Deshidrogenasa/metabolismo , Células Intersticiales del Testículo/patología , Masculino , Ratas , Ratas Wistar , Esteroides/biosíntesisRESUMEN
In the present study, it was hypothesized that the adrenocorticotrophin hormone receptor (ACTH-R) would be up-regulated in the adrenal gland of the sheep fetus following infusion of physiological amounts of ACTH, as shown for adrenal cortical cells in culture. In chronically catheterized sheep, an intravenous infusion of ACTH(1-24) was given to 6 fetuses for 24 h at a rate of 0.5 microg h(-1), starting on Day 126 or 127 of gestation (term approximately 147 days). Four control fetuses received an infusion of vehicle (saline). Total RNA was extracted from the fetal adrenal glands by the guanidinium thiocyanate method. Expression of specific mRNAs was determined by ribonuclease protection assay using cRNA probes directed against: ACTH-R; the steroid enzymes side-chain cleavage (P450scc), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), 17apha-hydroxylase (P450c17) and 21beta-hydroxylase (P450c21); and beta-actin. Ratios of mRNA expression to beta-actin mRNA expression (arbitrary units) were calculated to correct for differences in RNA quality between samples. The concentration (mean +/- SEM) of immunoreactive cortisol in fetal plasma was greater after ACTH infusion than after vehicle infusion (47 +/- 3 v. 13 +/- 2 ng mL(-1) respectively; P<0.001). Adrenal expression of P450scc and P450c21 mRNA increased after ACTH infusion (P<0.05), whereas expression of P450c17 and 3beta-HSD mRNA was unchanged. There was no difference in ACTH-R mRNA expression between ACTH- and vehicle-infused fetuses (254 +/- 48 v. 305 +/- 76 arbitrary units respectively). It was concluded that ACTH is able to increase plasma cortisol concentrations in the sheep fetus by up-regulating cortisol synthesis in the adrenal gland, but that in vivo this does not require up-regulation of ACTH-R mRNA.
Asunto(s)
Corteza Suprarrenal/embriología , Corteza Suprarrenal/fisiología , Hormona Adrenocorticotrópica/metabolismo , Receptores de Corticotropina/genética , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/genética , Equilibrio Ácido-Base , Corteza Suprarrenal/efectos de los fármacos , Hormona Adrenocorticotrópica/farmacología , Animales , Análisis de los Gases de la Sangre , Presión Sanguínea/efectos de los fármacos , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/efectos de los fármacos , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Sistema Enzimático del Citocromo P-450/efectos de los fármacos , Sistema Enzimático del Citocromo P-450/genética , Femenino , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Frecuencia Cardíaca Fetal/efectos de los fármacos , Hidrocortisona/sangre , Hidrocortisona/inmunología , Infusiones Intravenosas , Embarazo , ARN Mensajero/análisis , Receptores de Corticotropina/efectos de los fármacos , Receptores de Corticotropina/metabolismo , Reproducibilidad de los Resultados , Ovinos , Esteroide 17-alfa-Hidroxilasa/efectos de los fármacos , Esteroide 17-alfa-Hidroxilasa/genética , Esteroide 21-Hidroxilasa , Regulación hacia ArribaRESUMEN
Lead acetate (Pb) decreases the expression of steroidogenic acute regulatory (StAR) protein and the enzymatic activities of cytochrome P-450 side-chain cleavage (P450scc) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) in a concentration-dependent manner in Leydig cells at 2 h, the duration of submaximal inhibition. This study was undertaken at 3 h of Pb incubation to compare the effects at maximal metal inhibition of steroidogenesis. Quantitatively a 3-h Pb incubation with MA-10 cells resulted in higher decreases in human chorionic gonadotropin (hCG)-stimulated progesterone production, expression of StAR protein, and the activity of 3beta-HSD compared to 2 h. In contrast, lead inhibited dibutyryl cAMP (dbcAMP)-stimulated progesterone production but lacked this effect at 2 h. Surprisingly, Pb at 3 h of incubation did not affect P450scc enzyme activity, yet this enzymatic activity was inhibited at 2 h. Data indicate that incubation time is a factor in Pb-induced alterations in MA- 10 cell steroidogenesis.
Asunto(s)
Plomo/administración & dosificación , Plomo/toxicidad , Progesterona/biosíntesis , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Gonadotropina Coriónica/efectos de los fármacos , Gonadotropina Coriónica/metabolismo , Relación Dosis-Respuesta a Droga , Hidroxicolesteroles/farmacología , Immunoblotting , Plomo/farmacocinética , Tumor de Células de Leydig , Células Intersticiales del Testículo/efectos de los fármacos , Células Intersticiales del Testículo/metabolismo , Masculino , Ratones , Mitocondrias/metabolismo , Fosfoproteínas/efectos de los fármacos , Fosfoproteínas/metabolismo , Pregnenolona/farmacología , Radioinmunoensayo , Transducción de Señal/efectos de los fármacos , Factores de Tiempo , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Impaired testicular function was observed after an exposure of Swiss albino mice (30 +/- 2 g) to mercuric chloride. A sublethal chronic exposure (0.5 ppm for 21 days) resulted in regressed histological and histochemical properties of the testis. The changes observed were degenerated tunica albuginea, abnormal configurations of seminiferous tubules, deformed primary and secondary spermatocytes, hypertrophy and vacuolization in interstitial cells and Sertoli cells. The 3 beta and 17 beta hydroxy steroid dehydrogenase enzyme and the level of testosterone hormone were significantly (p < 0.001) reduced. The diameter of different spermatogenic cells were significantly (p < 0.001) reduced.
Asunto(s)
Cloruro de Mercurio/toxicidad , Espermatogénesis/efectos de los fármacos , Testículo/efectos de los fármacos , Contaminantes del Agua/toxicidad , 17-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , 3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , Animales , Masculino , Ratones , Testículo/citología , Testículo/enzimología , Testículo/patología , Testosterona/sangreRESUMEN
The molecular pathways involved in estrogen-mediated induction of hair growth in androgenetic alopecia are unknown. Some authors found that estradiol (E) inhibited 5alpha-reductase (5alpha-R) activity and therefore we addressed the question whether 17alpha- or 17beta-E are able to modulate the activity of 5alpha-R, 3beta-hydroxysteroid dehydrogenase (3beta-HSD) or 17beta-hydroxysteroid dehydrogenase (17beta-HSD) in isolated compartments of human hair follicles. For this purpose, scalp biopsies from volunteers were taken and from each biopsy root sheaths, connective tissue sheaths and dermal papillae (DP) were dissected and incubated in the presence of 3H-testosterone (T) and, in addition, either 17alpha-E, 17beta-E, progesterone or finasteride for up to 48 hrs. Thereafter high-performance liquid chromatography analysis of culture supernatants was performed to detect T-metabolites. At the tested concentrations, finasteride was found to be a major inhibitor of dihydrotestosterone (DHT) formation. Even 1 nM finasteride inhibited DHT synthesis in DP by 86% and 1 nM progesterone by 75%. Estrogens were less able to inhibit the synthesis of DHT in DP (e.g. 100 nM 17alpha-E: 20%; 100 nM 17beta-E: 60%). Whether E directly inhibits 5alpha-R in DP's or whether the effect of estrogens might be explained by an increased conversion of T to the weaker androgens such as androstendione (via 17beta-HSD), androstenediol (via 3beta-HSD) or 17beta-E (via aromatase), thereby diminishing the amount of T available for the conversion to DHT, remains to be shown.
Asunto(s)
3-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , Inhibidores de 5-alfa-Reductasa , Alopecia/tratamiento farmacológico , Alopecia/metabolismo , Dihidrotestosterona/antagonistas & inhibidores , Estradiol/farmacología , Finasterida/farmacología , Folículo Piloso/metabolismo , Progesterona/farmacología , Cromatografía Líquida de Alta Presión , Evaluación Preclínica de Medicamentos , Femenino , Folículo Piloso/crecimiento & desarrollo , Humanos , MasculinoRESUMEN
Lindane, the gamma isomer of hexachlorocyclohexane (HCH), is one of the oldest synthetic pesticides still in use worldwide. Numerous reports have shown that this pesticide adversely affects reproductive function in animals. Although the pathogenesis of reproductive dysfunction is not yet fully understood, recent reports indicate that lindane can directly inhibit adrenal and gonadal steroidogenesis. Because Leydig cells play a pivotal role in male reproductive function through the production of testosterone, the mouse MA-10 Leydig tumor cell line was used to assess the potential effects of gamma-HCH and its isomers, alpha-HCH and delta-HCH, on steroid production, steroidogenic enzyme expression and activity, and steroidogenic acute regulatory (StAR) protein expression. StAR mediates the rate-limiting and acutely regulated step in hormone-stimulated steroidogenesis, the intramitochondrial transfer of cholesterol to the P450(scc) enzyme. Our studies demonstrate that alpha-, delta-, and gamma-HCH inhibited dibutyryl ([Bu](2)) cAMP-stimulated progesterone production in MA-10 cells in a dosage-dependent manner without affecting general protein synthesis; and protein kinase A or steroidogenic enzyme expression, activity, or both. In contrast, each of these isomers dramatically reduced (Bu)(2)cAMP-stimulated StAR protein levels. Therefore, our results are consistent with the hypothesis that alpha-, delta-, and gamma-HCH inhibited steroidogenesis by reducing StAR protein expression, an action that may contribute to the pathogenesis of lindane-induced reproductive dysfunction.