RESUMEN
Most sheep are seasonal estrus, and they breed in autumn when the days get shorter. Seasonal estrus is an important factor that affects the productivity and fertility of sheep. The key point to solve this problem is to explore the regulation mechanism of estrus in sheep. Therefore, in this study, transcriptomic sequencing technology was used to identify differentially expressed mRNAs in the hypothalamus, pituitary and ovary of Small Tail Han sheep (year-round estrus) and tan sheep (seasonal estrus) among luteal, proestrus and estrus stages. There were 256,923,304,156 mRNAs being identified in the hypothalamus, pituitary and ovary, respectively. Functional analysis showed that the photosensor, leucine and isoleucine biosynthesis pathways were enriched significantly. It is speculated that photoperiod may initiate estrus by stimulating the corresponding pathways in hypothalamus. ODC1, PRLH, CRYBB2, SMAD5, OPN1SW, TPH1 are believed to be key genes involved in the estrogen process. In conclusion, this study expanded the database of indigenous sheep breeds, and also provided new candidate genes for future genetic and molecular studies on the seasonal estrus trait in sheep.
Asunto(s)
Estro/genética , Hipotálamo/metabolismo , Células Neuroendocrinas/metabolismo , Ovario/metabolismo , Hipófisis/metabolismo , Transcriptoma/genética , Anestro/genética , Anestro/metabolismo , Animales , Vías Biosintéticas/genética , Cruzamiento/métodos , Estrógenos/genética , Estrógenos/metabolismo , Estro/metabolismo , Femenino , Perfilación de la Expresión Génica/métodos , Isoleucina/genética , Isoleucina/metabolismo , Leucina/genética , Leucina/metabolismo , Fotoperiodo , ARN Mensajero/genética , Estaciones del Año , OvinosRESUMEN
OBJECTIVE: Previous study suggested that estradiol (E2) plays an important role in otolith shedding by regulating the expression of otoconin 90 (OC90). The purpose of this article is to provide further data on the effect and mechanism of E2 on the morphology of otolith. METHODS: The rats receiving bilateral ovariectomy (OVX) were used as animal models. Co-immunoprecipitation was used to observe the relationship between estrogen receptor (ER) and estrogen-related receptor α (ERRα). The morphology of otolith was observed under the scanning electron microscopy. Western blotting and qPCR were used for quantitative analysis of the roles of ER and ERRα in regulating OC90 expression. RESULTS: The looser otoliths were observed in rats receiving bilateral OVX, which could be reversed by supplementation with E2. The level of ERRα was decreased in bilateral OVX rats. ER and ERRα interacted with each other on the regulation of the expression of OC90. CONCLUSION: Our results suggest ER and ERRα are both important downstream receptors involved in regulating OC90 expression in utricles of rats, and ERRα probably functions by interacting with ER. This provides evidence for the mechanism of otolith shedding. And it may be significant for future studies of targeted prevention and therapies for benign paroxysmal positional vertigo.
Asunto(s)
Proteínas de Unión al Calcio/genética , Estrógenos/metabolismo , Membrana Otolítica/metabolismo , Receptores de Estrógenos/genética , Animales , Estradiol/metabolismo , Estrógenos/genética , Femenino , Humanos , Membrana Otolítica/patología , Ovariectomía , Ratas , Receptor Relacionado con Estrógeno ERRalfaRESUMEN
Sparassis crispa (Hanabiratake) is a widely used medicinal mushroom in traditional Chinese medicine because it contains materials with pharmacological activity. Here, we report its 39.0-Mb genome, encoding 13,157 predicted genes, obtained using next-generation sequencing along with RNA-seq mapping data. A phylogenetic analysis by comparison with 25 other fungal genomes revealed that S. crispa diverged from Postia placenta, a brown-rot fungus, 94 million years ago. Several features specific to the genome were found, including the A-mating type locus with the predicted genes for HD1 and HD2 heterodomain transcription factors, the mitochondrial intermediate peptidase (MIP), and the B-mating type locus with seven potential pheromone receptor genes and three potential pheromone precursor genes. To evaluate the benefits of the extract and chemicals from S. crispa, we adopted two approaches: (1) characterization of carbohydrate-active enzyme (CAZyme) genes and ß-glucan synthase genes and the clusters of genes for the synthesis of second metabolites, such as terpenes, indoles and polyketides, and (2) identification of estrogenic activity in its mycelial extract. Two potential ß-glucan synthase genes, ScrFKS1 and ScrFKS2, corresponding to types I and II, respectively, characteristic of Agaricomycetes mushrooms, were newly identified by the search for regions homologous to the reported features of ß-glucan synthase genes; both contained the characteristic transmembrane regions and the regions homologous to the catalytic domain of the yeast ß-glucan synthase gene FKS1. Rapid estrogenic cell-signaling and DNA microarray-based transcriptome analyses revealed the presence of a new category of chemicals with estrogenic activity, silent estrogens, in the extract. The elucidation of the S. crispa genome and its genes will expand the potential of this organism for medicinal and pharmacological purposes.
Asunto(s)
Genoma Fúngico/genética , Polyporales/genética , Transcriptoma/genética , Agaricales , Carbohidratos/genética , Mapeo Cromosómico , Estrógenos/genética , Glucosiltransferasas/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Análisis de Secuencia por Matrices de Oligonucleótidos , Filogenia , Polyporales/patogenicidad , Transducción de Señal , beta-Glucanos/metabolismoRESUMEN
Objectives: Menopausal transition in women initiates with declining estrogen levels and is followed by significant changes in their physiological characteristics. These changes often lead to medical conditions, such as obesity, which is correlated with chronic low-grade/subclinical inflammation. Ocimum gratissimum L. is a food spice or traditional herb in many countries; the plant is rich in antioxidants, which possess anti-inflammation activities and multitude of other therapeutic functions. Methods: In this study, we evaluated effects of O. gratissimum extract (OGE) in preventing obesity by using ovariectomized (OVX) animal models to mimic menopausal women. Methods: OVX rats showed increase in body weight and in adipocyte size in perigonadal adipose tissue (p <0.05) and decrease in uterus weight. By contrast, OGE (0.2 mg/ml) significantly reduced body weight gain and adipocyte in OVX rats and showed insignificant changes in uterus weight. Further investigation indicated that OGE exerted no influence on levels of dorsal fat, serum total cholesterol, and serum triacylglycerol and on serum biochemical factors, calcium, phosphorus, and glucose. Conclusion: These findings suggested that OGE dietary supplements may be useful in controlling body weight of menopausal women.
Asunto(s)
Obesidad/dietoterapia , Ocimum/química , Extractos Vegetales/administración & dosificación , Especias , Tejido Adiposo/efectos de los fármacos , Animales , Antioxidantes/administración & dosificación , Antioxidantes/química , Peso Corporal , Estrógenos/deficiencia , Estrógenos/genética , Femenino , Análisis de los Alimentos , Humanos , Menopausia/efectos de los fármacos , Obesidad/patología , Ovariectomía , Extractos Vegetales/química , Ratas , Útero/efectos de los fármacos , Útero/crecimiento & desarrolloRESUMEN
Kisspeptin (Kiss) plays a critical role in mediating gonadal steroid feedback to the gonadotropin-releasing hormone (GnRH) neurons in mammals. However, little information regarding the regulation of kisspeptin gene by sex steroids is available in teleosts. In this study, we examined the direct actions of estradiol (E2) and testosterone (T) on hypothalamic expression of kisspeptin and other key factors involved in reproductive function of half-smooth tongue sole. As a first step, a partial-length cDNA of kiss2 was identified from the brain of tongue sole and kiss2 transcript levels were shown to be widely expressed in various tissues, notably in the ovary. Then, the actions of sex steroids on kiss2 and other reproduction-related genes were evaluated using a primary hypothalamus culture system. Our results showed that neither kiss2 nor its receptor kiss2r mRNA levels were significantly altered by sex steroids. Moreover, sex steroids did not modify hypothalamic expression of gonadotropin-inhibitory hormone (gnih) and its receptor gnihr mRNAs, either. However, E2 markedly stimulated both gnrh2 and gnrh3 mRNAs levels. Overall, this study provides insights into the role of sex steroids in the reproductive function of Pleuronectiform teleosts.
Asunto(s)
Estrógenos/genética , Peces Planos/fisiología , Regulación de la Expresión Génica , Hormona Liberadora de Gonadotropina/genética , Hipotálamo/metabolismo , Kisspeptinas/genética , Reproducción/genética , Testosterona/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , ADN Complementario , Femenino , Filogenia , ARN Mensajero/genética , Homología de Secuencia de AminoácidoRESUMEN
Aromatase inhibitors (AIs) used as adjuvant therapy in postmenopausal women with hormone receptor-positive breast cancer cause diverse musculoskeletal side effects that include bone loss and its associated fracture. About half of the 391 patients treated with AIs in the Barcelona-Aromatase induced bone loss in early breast cancer cohort suffered a significant bone loss at lumbar spine (LS) and/or femoral neck (FN) after 2 years on AI-treatment. In contrast, up to one-third (19.6% LS, 38.6% FN) showed no decline or even increased bone density. The present study aimed to determine the genetic basis for this variability. SNPs in candidate genes involved in vitamin D and estrogen hormone-response pathways (CYP11A1, CYP17A1, HSD3B2, HSD17B3, CYP19A1, CYP2C19, CYP2C9, ESR1, DHCR7, GC, CYP2R1, CYP27B1, VDR and CYP24A1) were genotyped for association analysis with AI-related bone loss (AIBL). After multiple testing correction, 3 tag-SNPs (rs4077581, s11632698 and rs900798) located in the CYP11A1 gene were significantly associated (P<0.005) with FN AIBL at 2 years of treatment. Next, CYP11A1 expression in human fresh bone tissue and primary osteoblasts was demonstrated by RT-PCR. Both common isoforms of human cholesterol side-chain cleavage enzyme (encoded by CYP11A1 gene) were detected in osteoblasts by western blot. In conclusion, the genetic association of CYP11A1 gene with AIBL and its expression in bone tissue reveals a potential local function of this enzyme in bone metabolism regulation, offering a new vision of the steroidogenic ability of this tissue and new understanding of AI-induced bone loss.
Asunto(s)
Inhibidores de la Aromatasa/uso terapéutico , Densidad Ósea/efectos de los fármacos , Densidad Ósea/genética , Huesos/efectos de los fármacos , Huesos/metabolismo , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Densidad Ósea/fisiología , Huesos/fisiopatología , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Estrógenos/genética , Femenino , Genotipo , Humanos , Persona de Mediana Edad , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Osteoporosis Posmenopáusica/genética , Osteoporosis Posmenopáusica/metabolismo , Osteoporosis Posmenopáusica/fisiopatología , Polimorfismo de Nucleótido Simple/genética , Vitamina D/genéticaRESUMEN
We summarize updated information about DNA microarray-based gene expression profiling by focusing on its application to estrogenic chemicals. First, estrogenic chemicals, including natural/industrial estrogens and phytoestrogens, and the methods for detection and evaluation of estrogenic chemicals were overviewed along with a comprehensive list of estrogenic chemicals of natural or industrial origin. Second, gene expression profiling of chemicals using a focused microarray containing estrogen-responsive genes is summarized. Third, silent estrogens, a new type of estrogenic chemicals characterized by their estrogenic gene expression profiles without growth stimulative or inhibitory effects, have been identified so far exclusively by DNA microarray assay. Lastly, the prospect of a microarray assay is discussed, including issues such as commercialization, future directions of applications and quality control methods.
Asunto(s)
Disruptores Endocrinos/farmacología , Estrógenos/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Fitoquímicos/farmacología , Fitoestrógenos/farmacología , Línea Celular Tumoral , Estrógenos/genética , Humanos , Análisis por Micromatrices , Análisis de Secuencia por Matrices de Oligonucleótidos , Control de Calidad , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Transducción de SeñalRESUMEN
OBJECTIVE: To observe the effect of drug-paste separated moxibustion of "Mingmen" (GV 4) on the levels of serum estrogen (E2) and progesterone (P) and their endometrial receptor mRNA expression in rats with primary dysmenorrhea in order to investigate its mechanism underlying improvement of primary dysmenorrhea. METHODS: A total of 100 female SD rats were randomized into control, model, medication, acupuncture and moxibustion groups, with 20 rats in each group. Primary dysmenorrhea model was established by subcutaneous injection of Benzestrofol for 10 days and intraperitoneal injection of Oxytocin for 1 d. Rats of the medication group were fed with extractum leonuri inspissatum (8 g/100 g) and those of the moxibustion group treated with drug-paste separated moxibustion at "Mingmen" (GV 4). For rats of the acupuncture group, a filiform needle was inserted into GV 4, manipulated for a while and retained for 30 min. The treatment of the latter 3 groups was conducted once daily for 7 days. The rat's body-writhing latency and times during 30 min were recorded. The contents of serum E2 and P were detected by ELISA, and the expression of estrogen receptor (ER) mRNA and progesterone receptor (PR) mRNA in the endometrium was determined by quantitative real-time (RT)-PCR. RESULTS: (1) The body-writhing latency was shorter and the writhing times were more in the model group than in the control group (P < 0.01). Compared with the model group, the body-writhing latency was significantly increased and the writhing times were obviously decreased in the medication, acupuncture and moxibustion groups (P < 0.01). There were no significant differences among the medication, acupuncture and moxibustion groups in the body-writhing latency (P > 0.05), but the body-writhing numbers of the acupuncture and moxibustion groups were markedly lower than that of the medication group (P < 0.01). (2) Compared with the control group, serum E2 content and endometrial ER mRNA expression level were significantly increased, and serum P content and endometrial PR mRNA level evidently decreased in the model group (P < 0.01, P < 0.05). In comparison with the model group, serum E2 contents and endometrial ER mRNA expression levels were considerably down-regulated, and serum P contents and endometrial PR mRNA expression levels markedly up-regulated in the medication, acupuncture and moxibustion groups (P < 0.01, P < 0.05). The effects of the moxibustion group were significantly superior to those of the acupuncture and medication groups, and those of the acupuncture group were also significantly superior to those of the medication group in lowering E2 and endometrial ER mRNA levels, and raising serum P and endometrial PR mRNA expression levels (P < 0.01, P < 0.05). CONCLUSION: Drug-paste separated moxibustion of GV 4 is effective in relieving pain in primary dysmenorrheal rats, which is probably associated with its effects in down-regulating serum E2 content and endometrial ER mRNA expression, and up-regulating serum P and endometrial PR mRNA expression levels.
Asunto(s)
Dismenorrea/genética , Dismenorrea/terapia , Estrógenos/genética , Moxibustión , Progestinas/genética , Receptores de Progesterona/genética , Puntos de Acupuntura , Animales , Modelos Animales de Enfermedad , Dismenorrea/metabolismo , Endometrio/metabolismo , Estrógenos/metabolismo , Femenino , Humanos , Progestinas/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Receptores de Progesterona/metabolismoRESUMEN
Breast cancer is the most common cancer in women worldwide. There are many endocrine adjuvant therapies for breast cancer patients that are categorized according to their mechanisms. Among them, aromatase inhibitors (AIs) that block the synthesis of estrogens have proven superiority compared with tamoxifen and have replaced it as a first-line hormonal therapy. However, AIs also have limitations due to their side effects - increased rate of bone loss and musculoskeletal complaints. We therefore need new candidate AIs with fewer side effects. The extracts of Ginkgo biloba (EGb), which contain phytochemicals from the tree, had biphasic effects for estrogens and osteoporosis-inhibiting activities in our previous experiments. In this study, we explored the possibility of EGb as an AI and their mechanisms. Aromatase activities were inhibited by EGb both in JEG-3 cells and in recombinant CYP19 microsomes. The results of polymerase chain reaction for aromatase from a coding sequence and specific promoter sequences (exon I.a, exon I.c) in JEG-3 cells as well as the results of reporter gene assays showed that EGb dose-dependently decreased the aromatase gene expression. The decreased protein levels were demonstrated by Western blotting. From these results, we concluded that EGb could act as an AI at both the enzyme and transcriptional levels.
Asunto(s)
Inhibidores de la Aromatasa/farmacología , Aromatasa/metabolismo , Ginkgo biloba/química , Extractos Vegetales/farmacología , Línea Celular Tumoral , Coriocarcinoma/enzimología , Coriocarcinoma/patología , Estrógenos/genética , Estrógenos/farmacología , Estrógenos/uso terapéutico , Femenino , Humanos , Microsomas/efectos de los fármacos , Microsomas/enzimología , Regiones Promotoras Genéticas , Tamoxifeno/farmacología , Tamoxifeno/uso terapéutico , Neoplasias Uterinas/enzimología , Neoplasias Uterinas/patologíaRESUMEN
The fruits of Ligustrum lucidum (FLL) has long been used for the treatment of osteoporosis in China, but the antiosteoporotic compounds in FLL are still poorly understood. In this study, the alkaline phosphatase (ALP) activity-guided isolation of osteogenic components from FLL was carried out by using osteoblast-like UMR-106 cells. Eight compounds, namely tyrosol (1), tyrosyl acetate (2), hydroxytyrosol (3), salidroside (4), oleoside dimethyl ester (5), oleoside-7-ethyl-11-methyl ester (6), nuzhenide (7), and G13 (8), were isolated and identified. Further study showed that compounds 3, 4, 7, and 8 increased ALP activity in UMR-106 cells. Compounds 5, 6, and 7 promoted the proliferation of UMR-106 cells. The aqueous extract of FLL-activated ERα/ß-mediated gene transcription, whereas the isolated compounds were inactive. All eight isolated compounds also exhibited antioxidative activity, with compounds 1, 2, and 3 being the most potent. These results indicate that the antiosteoporotic effect of FLL is derived from different compounds together with different mechanisms such as ER-dependent or independent pathways and antioxidative effects. Salidroside (4) and nuzhenide (7) warrant further investigation as new pharmaceutical tools for the prevention and treatment of osteoporosis.
Asunto(s)
Fosfatasa Alcalina/metabolismo , Conservadores de la Densidad Ósea/farmacología , Frutas/química , Ligustrum/química , Osteoporosis/tratamiento farmacológico , Extractos Vegetales/farmacología , Animales , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Conservadores de la Densidad Ósea/aislamiento & purificación , Estrógenos/agonistas , Estrógenos/genética , Glucósidos/aislamiento & purificación , Glucósidos/farmacología , Células HeLa/efectos de los fármacos , Células HeLa/fisiología , Humanos , Osteoblastos/efectos de los fármacos , Osteoblastos/enzimología , Osteoporosis/enzimología , Fenoles/aislamiento & purificación , Fenoles/farmacología , Fitoterapia , Piranos/aislamiento & purificación , Piranos/farmacología , Ratas , Receptores de Estrógenos/genéticaRESUMEN
Sirtinol is a purported specific inhibitor of the nicotinamide adenine dinucleotide (NAD)-dependent type III histone deacetylase (also known as sirtuin). Sirtinol has been used extensively to identify chemopreventive/chemotherapeutic agents that modulate the sirtuins. However, the molecular effect of sirtinol other than serving as sirtuin inhibitor in cells is less clear. The present study addressed this deficiency in the literature. Based on structural similarity with plant-derived cancer preventive/therapeutic compounds such as 3', 3'-diindolylmethane, resveratrol, and genistein, we hypothesized that sirtinol may act on pathways similar to that affected by these compounds in the human prostate cancer cell LNCaP. We found that treatment of LNCaP cells with sirtinol led to concentration-dependent effects on multiple pathways. Sirtinol inhibited LNCaP cell cycle and growth that was correlated with up-regulation of cyclin-dependent kinase inhibitor 1A mRNA and protein levels. This effect of sirtinol may due in part to modulation of androgen, estrogen, and insulin-like growth factor-1 mediated pathways as sirtinol treatment led to inhibition of mRNA and protein expression of marker genes involved in these pathways. We also found sirtinol activates aryl hydrocarbon-dependent pathways in LNCaP cells. The effects of sirtinol were observed at 25 µM, a concentration lower than Ki (38 µM) for sirtuin activity. Based on these results we reasoned that sirtinol exerts pleiotropic effects in cells and that biological effects of sirtinol may not be due solely to inhibition of sirtuin.
Asunto(s)
Andrógenos/metabolismo , Benzamidas/farmacología , Naftoles/farmacología , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/metabolismo , Receptores Citoplasmáticos y Nucleares/metabolismo , Transducción de Señal/efectos de los fármacos , Sirtuinas/antagonistas & inhibidores , Andrógenos/genética , Anticarcinógenos/farmacología , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Estrógenos/genética , Estrógenos/metabolismo , Humanos , Factor I del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Masculino , NAD(P)H Deshidrogenasa (Quinona)/genética , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Fitoterapia , Neoplasias de la Próstata/genética , Receptores Citoplasmáticos y Nucleares/genética , Transducción de Señal/genética , Sirtuinas/genética , Sirtuinas/metabolismo , Regulación hacia Arriba/efectos de los fármacosRESUMEN
The mechanisms through which estradiol (E2) regulates gonadotropin-releasing hormone (GnRH) neurons to control fertility are unclear. Previous studies have demonstrated that E2 rapidly phosphorylates cAMP response element-binding protein (CREB) in GnRH neurons in vivo. In the present study, we used GnRH neuron-specific CREB-deleted mutant mice [GnRH-CREB knock-outs (KOs)] with and without global cAMP response element modulator (CREM) deletion (global-CREM KOs) to investigate the role of CREB in estrogen negative feedback on GnRH neurons. Evaluation of GnRH-CREB KO mice with and without global CREM deletion revealed normal puberty onset. Although estrus cycle length in adults was the same in controls and knock-out mice, cycles in mutant mice consisted of significantly longer periods of diestrus and less estrus. In GnRH-CREB KO mice, basal levels of luteinizing hormone (LH) and the postovariectomy increment in LH were normal, but the ability of E2 to rapidly suppress LH was significantly blunted. In contrast, basal and postovariectomy LH levels were abnormal in GnRH-CREB KO/global-CREM KO mice. Fecundity studies showed that GnRH-CREB KO with and without global CREM deletion were normal up to â¼9 months of age, at which time they became prematurely reproductively senescent. Morphological analysis of GnRH neurons revealed a significant reduction (p < 0.01) in GnRH somatic spine density of GnRH-CREB KO mice compared to control females. These observations implicate CREB within the GnRH neuron as an important target for E2's negative feedback actions. They also indicate that the rapid modulation of CREB by E2 is of physiological significance in the CNS.
Asunto(s)
Proteína de Unión a CREB/metabolismo , Estrógenos/metabolismo , Retroalimentación Fisiológica/fisiología , Hormona Liberadora de Gonadotropina/metabolismo , Neuronas/metabolismo , Envejecimiento/efectos de los fármacos , Envejecimiento/genética , Envejecimiento/metabolismo , Análisis de Varianza , Animales , Proteína de Unión a CREB/deficiencia , Modulador del Elemento de Respuesta al AMP Cíclico/deficiencia , Modulador del Elemento de Respuesta al AMP Cíclico/metabolismo , Espinas Dendríticas/metabolismo , Estradiol , Estrógenos/genética , Ciclo Estral/genética , Femenino , Fertilidad/genética , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Hormona Liberadora de Gonadotropina/deficiencia , Hipotálamo/citología , Hormona Luteinizante/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mutación/genética , Neuronas/ultraestructura , Ovariectomía , RadioinmunoensayoRESUMEN
Degradation of heavy oil by bacteria to decompose organic compounds such as aliphatic and aromatic hydrocarbons has been used in bioremediation. However, the biological and environmental effects of the degradation products including intermediates are still not clear. Here, we monitored the degradation of C-heavy oil by analyzing the products formed in cultures with oil-degrading bacteria (complex microbes or a single bacterial strain). Furthermore, proliferation assays using breast cancer MCF-7 cells and gene-expression profiling of MCF-7 cells using oligonucleotide-DNA microarrays were performed to evaluate the estrogenic activity of the degradation products. While the products did not show any significant cell-proliferative activity, the oil samples cultured for longer periods (2-3 months), whether cultured with mixed microbes or a single bacterial strain, showed gene-expression profiles similar to that of 17ß-estradiol (E2). These results suggest that oil-degradation products have estrogenic activity, and estrogen-like components could possibly be produced during the degradation process.
Asunto(s)
Estradiol/genética , Estrógenos/genética , Contaminación por Petróleo , Petróleo/toxicidad , Contaminantes Químicos del Agua/toxicidad , Biodegradación Ambiental , Línea Celular Tumoral , Estradiol/metabolismo , Estrógenos/metabolismo , Estrógenos/toxicidad , Perfilación de la Expresión Génica , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Petróleo/metabolismo , Contaminantes Químicos del Agua/metabolismoRESUMEN
We previously demonstrated the protective effects of blueberry (BB) and black raspberry (BRB) supplemented at 2.5% dose in an ACI rat mammary tumor model. Here, we assessed a dose-related alteration in tumor indices with diet supplemented with 5% BB or BRB powder. The diet was well tolerated. Tumor palpation from 12 weeks revealed first tumor appearance by 84 days in the control group, that was delayed by 24 and 39 days with the BB and BRB diets, respectively (p = 0.04). Ellagic acid detected in the plasma of rats fed the BRB diet was in the range of 96.6-294.2 ng/mL. While the BB diet showed better efficacy in reducing mammary tissue proliferation and tumor burden, tumor latency was delayed efficiently by BRB. Furthermore, BB was effective in downregulating CYP1A1 expression, while BRB downregulated ERα expression effectively. Distinct anticarcinogenic effects of the two berries correspond to their distinct phytochemical signatures.
Asunto(s)
Antineoplásicos Fitogénicos/administración & dosificación , Arándanos Azules (Planta)/química , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Estrógenos/metabolismo , Extractos Vegetales/administración & dosificación , Rosaceae/química , Animales , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Transformación Celular Neoplásica/efectos de los fármacos , Estrógenos/genética , Femenino , Frutas/química , Humanos , Ratas , Ratas Endogámicas ACIRESUMEN
Due to the health risks attributed to perimenopausal hormone therapy, phytoestrogens such as flavonoids are receiving widespread attention to help alleviate menopausal symptoms, including hormone-driven mood disorders. Based on our previous reporter gene study regarding their transactivational activity in raphe nuclei cells from a brain region involved in regulation of mood disturbances, we herein study their effects on the regulation of expression of 17ß-estradiol (E2)-regulated genes. DNA microarray was used to globally assess E2-induced gene expression in RNDA cells, a rat raphe nuclei-derived cellular model expressing oestrogen receptor ß. Out of 212 regulated genes, six were selected for verification and as endpoints for the effect of flavonoids on the regulation of mRNA expression in proliferating as well as differentiating RNDA cells. Under proliferative conditions, E2 up-regulated mRNA expression of Cml-5, Sox-18 and Krt-19. Similar effects were observed in response to 8-prenylnaringenin (8-PN), genistein (GEN), daidzein (DAI) and equol (EQ). In line with E2, mRNA expression of Nefm and Zdhhc-2 was down-regulated following 8-PN, GEN, DAI, EQ and naringenin treatment. No regulation was observed on Slc6a4 mRNA expression in response to E2 or the flavonoids in proliferating RNDA cells. When cells were shifted to conditions promoting differentiation, changes in cell morphology, in mRNA expression levels and in responsiveness towards E2 and the tested flavonoids were noticed. These expression studies additionally highlighted some of the genes as markers for RNDA cellular differentiation. RNDA cells should prove useful to elucidate molecular and cellular mechanisms of exogenous oestrogen receptor ligands with neural cell populations.
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Estradiol/farmacología , Receptor beta de Estrógeno/genética , Estrógenos/biosíntesis , Flavonoides/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Animales , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Línea Celular , Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Equol/farmacología , Estrógenos/genética , Flavanonas/farmacología , Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica/genética , Genisteína/farmacología , Isoflavonas/farmacología , Fitoestrógenos/farmacología , ARN Mensajero/genética , Ratas , Factores de Transcripción SOXF/genéticaRESUMEN
We and others have reported that mouse embryonic testes contain a subpopulation of somatic cells expressing estrogen receptor alpha (ERalpha). In order to provide evidence for a possible direct estrogen effect on mammalian testes from the early stage of their differentiation, here we devised a method for the in vitro culture of the ERalpha-expressing cells from 12.5 days post coitum mouse testes and their transfection with plasmids containing the classical estrogen responsive element (ERE) or the alternative estrogen AP-1 responsive element upstream of the luciferase reporter gene (ERE-Luc and AP-1-Luc). StAR immunopositivity of the most part of the ERalpha+ cells grown in culture and subjected to the estrogenic assay, allowed their identification as embryonic Leydig cells. Maximum induction of the ERE-Luc activity was achieved with 10 nM 17-beta estradiol (E2), from 1.7 to 3-fold in such cells and from 2.3 to 5.7-fold in MCF-7 cells used for comparison; the anti-estrogen ICI 182.780 abolished such effects. AP-1-Luc was less sensitive to E2 in both cell types (10 nM E2, 1.2 to 2.7-fold increase in embryonic Leydig cells; about 3-fold in MCF-7 cells) and the effect was not ICI-dependent. Eventually, we stimulated the transfected cells with various xenoestrogens such as lindane, bisphenol A or mono-(2-ethylhexyl) pthalate and with the phytoestrogen zeralenone obtaining evidence for ERE-Luc, but not AP-1-Luc stimulation in embryonic Leydig cells. These results represent evidence of functional ERalpha-dependent genomic pathways in embryonic Leydig cells and describe an in vitro assay suitable for evaluating the activity of putative estrogenic compounds on such cells.
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Bioensayo , Antagonistas de Estrógenos/farmacología , Estrógenos/farmacología , Células Intersticiales del Testículo/metabolismo , Testículo/embriología , Animales , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Células Cultivadas , Estradiol/análogos & derivados , Receptor alfa de Estrógeno/genética , Receptor alfa de Estrógeno/metabolismo , Receptor alfa de Estrógeno/fisiología , Estrógenos/genética , Estrona/genética , Femenino , Fulvestrant , Genes Reporteros/efectos de los fármacos , Humanos , Luciferasas/genética , Masculino , Ratones , Fitoestrógenos/farmacología , Elementos de Respuesta/genética , Factor de Transcripción AP-1/genética , Factor de Transcripción AP-1/metabolismo , TransfecciónRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Our previous work has demonstrated that several plants in the Piperaceae family are commonly used by the Q'eqchi Maya of Livingston, Guatemala to treat amenorrhea, dysmenorrhea, and pain. Extracts of Piper hispidum Swingle (Piperaceae), bound to the estrogen (ER) and serotonin (5-HT7) receptors. AIM OF THE STUDY: To investigate the estrogenic and serotonergic activities of Piper hispidum extracts in functionalized assays, identify the active chemical constituents in the leaf extract, and test these compounds as agonists or antagonists of ER and 5-HT7. MATERIALS AND METHODS: The effects of the Piper hispidum leaf extracts were investigated in estrogen reporter gene and endogenous gene assays in MCF-7 cells to determine if the extracts acted as an estrogen agonist or antagonist. In addition, the active compounds were isolated using ER- and 5-HT7 receptor bioassay-guided fractionation. The structures of the purified compounds were identified using high-resolution LC-MS and NMR spectroscopic methods. The ER- and 5-HT7-agonist effects of the purified chemical constituents were tested in a 2ERE-reporter gene assay in MCF-7 cells and in serotonin binding and functionalized assays. RESULTS: Three butenolides including one new compound (1) were isolated from the leaves of Piper hispidum, and their structures were determined. Compound 1 bound to the serotonin receptor 5-HT(7) with IC(50) values of 16.1 and 8.3 microM, respectively, and using GTP shift assays, Compound 1 was found to be a partial agonist of the 5-HT(7) receptor. The Piper hispidum leaf extracts, as well as Compounds 2 and 3 enhanced the expression of estrogen responsive reporter and endogenous genes in MCF-7 cells, demonstrating estrogen agonist effects. CONCLUSIONS: Extracts of Piper hispidum act as agonists of the ER and 5-HT(7) receptors. Compound 1, a new natural product, identified as 9,10-methylenedioxy-5,6-Z-fadyenolide, was isolated as the 5-HT(7) agonist. Compounds 2 and 3 are reported for the first time in Piper hispidum, and identified as the estrogen agonists. No inhibition of CYP450 was observed for any of these compounds in concentrations up to 1 microM. These activities are consistent with the Q'eqchi traditional use of the plant for the treatment of disorders associated with the female reproductive cycle.
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4-Butirolactona/análogos & derivados , Estrógenos/metabolismo , Fitoestrógenos/farmacología , Piperaceae/química , Extractos Vegetales/farmacología , Receptores de Serotonina/metabolismo , Agonistas de Receptores de Serotonina/farmacología , 4-Butirolactona/química , 4-Butirolactona/aislamiento & purificación , 4-Butirolactona/farmacología , Estrógenos/genética , Expresión Génica/efectos de los fármacos , Genes Reporteros , Humanos , Concentración 50 Inhibidora , Estructura Molecular , Fitoestrógenos/aislamiento & purificación , Extractos Vegetales/química , Hojas de la Planta , Agonistas de Receptores de Serotonina/aislamiento & purificaciónRESUMEN
OBJECTIVE: To observe the effect of preventive acupuncture and moxibustion of "Guanyuan" (CV 4) on uterus in ovariectomized (OVX) rats. METHODS: A total of 80 female SD rats were randomly divided into 5 groups: normal control, sham-operation (sham), model, preventive acupuncture (PA) and preventive moxibustion (PM) groups, with 16 cases in each. PA (with the needle retained for 20 min after insertion) and PM (one moxa-cone/rat) were applied to "Guanyuan" (CV 4) separately before ovariectomy, two times a week, 4 weeks altogether. Then, ovariectomy was performed on rats of model, PA and PM groups. Uterus tissue was taken under anesthesia for homogenate (10 rats/group) or sectioning (5-6 microm, 6 rats/group). Uterus estradiol (E2), progestone (P) were detected with radioimmunoassay; superoxide dismutase (SOD), nitric oxide synthase (NOS), and malondialdehyde (MDA) were assayed with immunoturdidimetry. Partial uterus slices were stained with H & E method for observing morphological changes, or stained with immunohistochemical method or with in situ hybridization method for displaying estrogen receptor (ER)-alpha and ER-alpha mRNA expression separately. RESULTS: After ovariectomy, the rat's uterus presented obvious squamous metaplasia, hyperplasy, and thickening of the endomembrane, decrease in glands and blood vessels and increase in fibrous connective tissue, etc; while the situation was evidently better in PA and PM groups. In comparison with normal control group, E2, P contents and SOD, NOS activities of uterus tissue in model group decreased significantly (P < 0.01), while MDA in model group increased evidently (P < 0.01). Compared with model group, uterus P content in PA and PM groups increased obviously (P < 0.05), while uterus MDA decreased apparently (P < 0.01). Compared with normal control group, the expression of ER-alpha and ER-alpha mRNA in model group was downregulated markedly (P < 0.01); after PA and PM, both ER-alpha and ER-alpha mRNA expression increased obviously (P < 0.01). No significant differences were found between PA and PM groups in the above mentioned 7 indexes (P > 0.05). CONCLUSION: Preventive acupuncture and moxibustion of CV4 can postpone the structural degeneration of uterus in OVX rats, which may be related to their effects in modulating the secretion of uterus E2 and P, upregulating the expression of ER-a and ER-alpha mRNA, and improving the anti-oxidative ability.
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Terapia por Acupuntura , Ovariectomía , Complicaciones Posoperatorias/prevención & control , Útero/metabolismo , Útero/cirugía , Animales , Estrógenos/genética , Estrógenos/metabolismo , Femenino , Expresión Génica , Humanos , Moxibustión , Óxido Nítrico Sintasa/genética , Óxido Nítrico Sintasa/metabolismo , Complicaciones Posoperatorias/metabolismo , Complicaciones Posoperatorias/terapia , Progesterona/genética , Progesterona/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Útero/enzimologíaRESUMEN
In order to evaluate estrogenic compounds in natural products, an in vitro detection system was established. For this system, the human breast cancer cell line MCF7 was stably transfected using an estrogen responsive chloramphenicol acetyltransferase (CAT) reporter plasmid yielding MCF7/pDsCAT-ERE119-Ad2MLP cells. To test the estrogenic responsiveness of this in vitro assay system, MCF7/pDsCAT-ERE119-Ad2MLP cells were treated with various concentrations of 17beta-estradiol. Treatments of 10(-8) to 10(-12) M 17beta-estradiol revealed significant concentration dependent estrogenic activities compared with ethanol. We used in vitro assay system to detect estrogenic effects in Puerariae radix and Ginseng radix Rubra extracts. Treatment of 500 and 50 microg/ml of Puerariae radix extracts increased the transcriptional activity approximately 4- and 1.5-fold, respectively, compared with the ethanol treatment. Treatment of 500, 50, and 5 microg/ml of Ginseng radix Rubra extracts increased the transcriptional activity approximately 3.2-, 2.7-, and 1.4-fold, respectively, compared with the ethanol treatment. These observations suggest that Puerariae radix and Ginseng radix Rubra extracts have effective estrogenic actions and that they could be developed as estrogenic supplements.