RESUMEN
Urine contains multiple water-soluble hormones, which are valuable non-invasive biomarkers for the monitoring of reproductive status and health. An effective method for drying urine on filter paper was previously developed to preserve wildlife urine samples where electrical equipment was not available for this; however, the stability of samples preserved in this way remains to be verified. Here, we developed and validated a method to elute multiple water-soluble reproductive hormones from filter paper that had been stored for an extended period of time. Aliquots of urine from chimpanzees were adsorbed on filter papers, air dried and stored for 1 year at room temperature. Estrone-3-conjugate (E1C), pregnanediol-3-glucuronide (PdG), estriol-3-glucuronide (E3G), and chorionic gonadotropin (CG) were eluted into deionized water from the filter papers and measured using enzyme immunoassays (EIAs). The mean recoveries of E1C, PdG, and creatinine from filter papers stored for 1 year were 69.5%, 128.7%, and 83.8%, respectively. The profiles of E1C and PdG from preserved filter papers significantly correlated with those derived from a direct analysis of the frozen urine of menstruating chimpanzees. We detected E3G and CG from 1-year-old filter papers for urine collected during early pregnancy, but the recovery of E3G was low and CG profiles did not correlate with those of the original frozen urine samples. The method proposed here for the elution and measurement of reproductive hormones in urine preserved for a long period of time on filter paper provides a practical and simple way to monitor the reproductive status of chimpanzees. We propose that this method can also be utilized in field studies of other wild nonhuman primates.
Asunto(s)
Gonadotropina Coriónica/análisis , Estriol/análogos & derivados , Pan troglodytes/orina , Pregnanodiol/análogos & derivados , Animales , Gonadotropina Coriónica/orina , Estriol/análisis , Estriol/orina , Femenino , Técnicas para Inmunoenzimas/métodos , Ciclo Menstrual/orina , Pan troglodytes/fisiología , Papel , Pregnanodiol/análisis , Pregnanodiol/orina , Manejo de Especímenes/métodosRESUMEN
Growth and development of an embryo or fetus during human pregnancy mainly depend on intact hormone biosynthesis and metabolism in maternal amniotic fluid (AF). We investigated the hormonal milieu in AF and developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of 14 sulfated and 6 unconjugated steroids in AF. 65â¯Aâ¯F samples (male: femaleâ¯=â¯35: 30) of mid-gestation ranging from 16th week of gestation to 25th week of gestation were analyzed. Reference data of 20 steroid levels in AF of healthy women were provided. 13 sulfated and 3 unconjugated steroids were for the first time quantified in AF by LC-MS/MS. Highest concentrations were found for pregnenolone sulfate (PregS: mean⯱â¯SD, 8.6⯱â¯3.7â¯ng/mL), 17α-hydroxypregnenolone sulfate (17OHPregS: 4.9⯱â¯2.0â¯ng/mL), epitestosterone sulfate (eTS: 7.3⯱â¯3.6â¯ng/mL), 16α-hydroxydehydroepiandrosterone sulfate (16OH-DHEAS: 21.5⯱â¯10.7â¯ng/mL), androsterone sulfate (AnS: 9.2⯱â¯7.4â¯ng/mL), estrone sulfate (E1S: 3.0⯱â¯3.0â¯ng/mL), estriol 3-sulfate (E3S: 8.1⯱â¯4.0â¯ng/mL) and estriol (E3: 1.2⯱â¯0.4â¯ng/mL). Only testosterone (T) showed a significant sex difference (pâ¯<â¯0.0001). Correlations between AF steroids mirrored the steroid metabolism of the feto-placental unit, and not only confirmed the classical steroid pathway, but also pointed to a sulfated steroid pathway.
Asunto(s)
Líquido Amniótico/química , Segundo Trimestre del Embarazo/fisiología , Esteroides/análisis , 17-alfa-Hidroxipregnenolona/análisis , Androsterona/análisis , Cromatografía Liquida , Deshidroepiandrosterona/análisis , Epitestosterona/análisis , Estriol/análogos & derivados , Estriol/análisis , Estrona/análogos & derivados , Estrona/análisis , Femenino , Edad Gestacional , Humanos , Masculino , Embarazo , Pregnenolona/análisis , Espectrometría de Masas en TándemRESUMEN
BACKGROUND: Although diabetic retinopathy (DR) has long been considered as a microvascular disorder, mounting evidence suggests that diabetic retinal neurodegeneration, in particular synaptic loss and dysfunction of retinal ganglion cells (RGCs) may precede retinal microvascular changes. Key molecules involved in this process remain poorly defined. The microtubule-associated protein tau is a critical mediator of neurotoxicity in Alzheimer's disease (AD) and other neurodegenerative diseases. However, the effect of tau, if any, in the context of diabetes-induced retinal neurodegeneration has yet to be ascertained. Here, we investigate the changes and putative roles of endogeneous tau in diabetic retinal neurodegeneration. METHODS: To this aim, we combine clinically used electrophysiological techniques, i.e. pattern electroretinogram and visual evoked potential, and molecular analyses in a well characterized high-fat diet (HFD)-induced mouse diabetes model in vivo and primary retinal ganglion cells (RGCs) in vitro. RESULTS: We demonstrate for the first time that tau hyperphosphorylation via GSK3ß activation causes vision deficits and synapse loss of RGCs in HFD-induced DR, which precedes retinal microvasculopathy and RGCs apoptosis. Moreover, intravitreal administration of an siRNA targeting to tau or a specific inhibitor of GSK3ß reverses synapse loss and restores visual function of RGCs by attenuating tau hyperphosphorylation within a certain time frame of DR. The cellular mechanisms by which hyperphosphorylated tau induces synapse loss of RGCs upon glucolipotoxicity include i) destabilizing microtubule tracks and impairing microtubule-dependent synaptic targeting of cargoes such as mRNA and mitochondria; ii) disrupting synaptic energy production through mitochondria in a GSK3ß-dependent manner. CONCLUSIONS: Our study proposes mild retinal tauopathy as a new pathophysiological model for DR and tau as a novel therapeutic target to counter diabetic RGCs neurodegeneration occurring before retinal vasculature abnormalities.
Asunto(s)
Estriol/análogos & derivados , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Mitocondrias/metabolismo , Fosforilación/fisiología , Tauopatías/metabolismo , Enfermedad de Alzheimer/metabolismo , Animales , Modelos Animales de Enfermedad , Estriol/metabolismo , Potenciales Evocados Visuales/fisiología , Masculino , Ratones Endogámicos C57BL , Retina/metabolismo , Células Ganglionares de la Retina/metabolismo , Proteínas tau/metabolismoRESUMEN
Microglia, traditionally known as resident immune cells in the brain, have been recently identified as one of the important cell types engaging in the formation and maintenance of neural circuitry, especially during the development. In this review, I describe the diversity of microglial functions revealed in different phases of development, ranging from neurogenesis to circuit formation. In particular, microglia possess dual functions in supporting the survival of neuronal structures or neural cells themselves, and removing excess neural components. Understanding these processes and underlying molecular mechanisms will provide important insights into the regulation of neural circuitry development. It further implicates the involvement of microglial dysfunction in neurodevelopmental disorders.
Asunto(s)
Encéfalo , Microglía , Animales , Encéfalo/citología , Encéfalo/crecimiento & desarrollo , Encéfalo/metabolismo , Movimiento Celular , Enfermedades del Sistema Nervioso Central/metabolismo , Estriol/análogos & derivados , Humanos , Microglía/citología , Microglía/metabolismoRESUMEN
Estrogens are potent and efficacious neuroprotectants both in vitro and in vivo in a variety of models of neurotoxicity. We determined the structural requirements for neuroprotection in an in vitro assay using a panel of >70 novel estratrienes, synthesized to reduce or eliminate estrogen receptor (ER) binding. We observed that neuroprotection could be enhanced by as much as 200-fold through modifications that positioned a large bulky group at the C2 or C4 position of the phenolic A ring of the estratriene. Further, substitutions on the B, C or D rings either reduced or did not markedly change neuroprotection. Collectively, there was a negative correlation between binding to ERs and neuroprotection with the more potent compounds showing no ER binding. In an in vivo model for neuroprotection, transient cerebral ischemia, efficacious compounds were active in protection of brain tissue from this pro-oxidant insult. We demonstrated that these non-feminizing estrogens engage in a redox cycle with glutathione, using the hexose monophosphate shunt to apply cytosolic reducing potential to cellular membranes. Together, these results demonstrate that non-feminizing estrogens are neuroprotective and protect brain from the induction of ischemic- and Alzheimer's disease (AD)-like neuropathology in an animal model. These features of non-feminizing estrogens make them attractive compounds for assessment of efficacy in AD and stroke, as they are not expected to show the side effects of chronic estrogen therapy that are mediated by ER actions in the liver, uterus and breast.
Asunto(s)
Encéfalo/efectos de los fármacos , Estradiol/farmacología , Estriol/farmacología , Estrógenos/farmacología , Neuronas/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Animales , Encéfalo/metabolismo , Encéfalo/patología , Estradiol/análogos & derivados , Estradiol/química , Estradiol/metabolismo , Estriol/análogos & derivados , Estriol/química , Estriol/metabolismo , Estrógenos/química , Estrógenos/metabolismo , Humanos , Estructura Molecular , Neuronas/metabolismo , Neuronas/patología , Fármacos Neuroprotectores/química , Fármacos Neuroprotectores/metabolismo , Oxidación-Reducción , Estrés Oxidativo/efectos de los fármacos , Receptores de Estrógenos/metabolismo , Relación Estructura-ActividadRESUMEN
The fusion of neurotransmitter-filled vesicles during synaptic transmission is balanced by endocytotic membrane retrieval. Despite extensive research, the speed and mechanisms of synaptic vesicle endocytosis have remained controversial. Here, we establish low-noise time-resolved membrane capacitance measurements that allow monitoring changes in surface membrane area elicited by single action potentials and stronger stimuli with high-temporal resolution at physiological temperature in individual bona-fide mature central synapses. We show that single action potentials trigger very rapid endocytosis, retrieving presynaptic membrane with a time constant of 470 ms. This fast endocytosis is independent of clathrin but mediated by dynamin and actin. In contrast, stronger stimuli evoke a slower mode of endocytosis that is clathrin, dynamin, and actin dependent. Furthermore, the speed of endocytosis is highly temperature dependent with a Q10 of â¼3.5. These results demonstrate that distinct molecular modes of endocytosis with markedly different kinetics operate at central synapses.
Asunto(s)
Potenciales de Acción/fisiología , Clatrina/metabolismo , Endocitosis/fisiología , Estriol/análogos & derivados , Animales , Estriol/fisiología , Hipocampo/metabolismo , Ratones Endogámicos C57BL , Ratas , Transmisión Sináptica/fisiología , Temperatura , Factores de TiempoRESUMEN
Glutamate is the main excitatory neurotransmitter in the central nervous system and plays an important role in most aspects of normal brain function. In spite of its importance as a neurotransmitter, excess glutamate is toxic to neurons. Clearance of extracellular glutamate is critical for maintenance of low extracellular glutamate concentration, and occurs in large part through the activity of GLT1 (EAAT2) and GLAST (EAAT1), which are primarily expressed by astrocytes. Rare variants and down-regulation of GLT1 and GLAST, in psychiatric disorders have been reported. In this review, we demonstrate that various kinds of GLT1 and/or GLAST knockout mice replicate many aspects of the behavioral abnormalities seen in major mental illnesses including schizophrenia, depression, obsessive -compulsive disorders, autism, epilepsy and addiction.
Asunto(s)
Sistema de Transporte de Aminoácidos X-AG/metabolismo , Trastornos Mentales/metabolismo , Factores de Edad , Animales , Astrocitos/metabolismo , Estriol/análogos & derivados , Estriol/metabolismo , Humanos , Factores de RiesgoRESUMEN
Human genetic studies have identified the neuronal RNA binding protein, Rbfox1, as a candidate gene for autism spectrum disorders. While Rbfox1 functions as a splicing regulator in the nucleus, it is also alternatively spliced to produce cytoplasmic isoforms. To investigate the function of cytoplasmic Rbfox1, we knocked down Rbfox proteins in mouse neurons and rescued with cytoplasmic or nuclear Rbfox1. Transcriptome profiling showed that nuclear Rbfox1 rescued splicing changes, whereas cytoplasmic Rbfox1 rescued changes in mRNA levels. iCLIP-seq of subcellular fractions revealed that Rbfox1 bound predominantly to introns in nascent RNA, while cytoplasmic Rbox1 bound to 3' UTRs. Cytoplasmic Rbfox1 binding increased target mRNA stability and translation, and Rbfox1 and miRNA binding sites overlapped significantly. Cytoplasmic Rbfox1 target mRNAs were enriched in genes involved in cortical development and autism. Our results uncover a new Rbfox1 regulatory network and highlight the importance of cytoplasmic RNA metabolism to cortical development and disease.
Asunto(s)
Trastorno Autístico/genética , Estriol/análogos & derivados , Neuronas/metabolismo , ARN Mensajero/metabolismo , Proteínas de Unión al ARN/genética , Empalme Alternativo/genética , Animales , Encéfalo/crecimiento & desarrollo , Encéfalo/metabolismo , Citoplasma/metabolismo , Estriol/metabolismo , Perfilación de la Expresión Génica/métodos , Humanos , Ratones Endogámicos C57BL , Factores de Empalme de ARNAsunto(s)
Encefalitis/inmunología , Enfermedad de Hashimoto/inmunología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antígenos/inmunología , Autoanticuerpos/inmunología , Niño , Preescolar , Encefalitis/patología , Estriol/análogos & derivados , Estriol/inmunología , Femenino , Enfermedad de Hashimoto/patología , Humanos , Lactante , Masculino , Persona de Mediana Edad , Canales de Potasio con Entrada de Voltaje/inmunología , Receptores de Glutamato/inmunología , Adulto JovenAsunto(s)
Autoanticuerpos/inmunología , Encefalitis Límbica/inmunología , Canales de Potasio con Entrada de Voltaje/inmunología , Epilepsia/etiología , Estriol/análogos & derivados , Estriol/inmunología , Humanos , Encefalitis Límbica/complicaciones , Encefalitis Límbica/epidemiología , Encefalitis Límbica/patología , PronósticoRESUMEN
Heng-Gu-Gu-Shang-Yu-He-Ji, also known as OsteoKing, is used as a herbal Traditional Chinese Medicine for the treatment of bone disease, including femoral head necrosis and osteoarthritis. However, whether OsteoKing has anti-osteoporotic properties has remained to be elucidated. The purpose of the present study was therefore to investigate the effects of OsteoKing on ovariectomy-induced osteoporosis in rabbits. Female New Zealand white rabbits were randomly divided into an ovariectomized (OVX) group and a sham-surgery group. The rabbits in the OVX group were subjected to an ovariectomy, while the rabbits in the sham group were subjected to the removal of an area of fat near the two ovaries. Bone mineral density, mechanical properties, serum biochemical parameters and micro-architecture were examined at 150 days post-OVX to characterize the experimental animal model. Once the osteoporotic rabbit model had been established, the rabbits in the OVX group were divided into the following groups: Model group, nilestriol group and 300 and 600 mg/kg OsteoKing groups, containing 16 rabbits in each group. OsteoKing and nilestriol were administered orally. The bone mineral density, mechanical properties, serum biochemical parameters, histology and micro-architecture were examined using dual-energy X-ray absorptiometric analysis, mechanical assessments, enzyme-linked immunosorbent assays, histopathological evaluation and micro-computerized tomography examination following 60 days and 120 days of treatment, respectively. Treatment with OsteoKing led to an elevation in the bone mineral density of the vertebra and serum phosphorus levels, reduced serum concentrations of osteocalcin, procollagen type I N-terminal peptide, tartrate-resistant acid phosphatase 5b and cross-linked N-telopeptide of type I collagen, improved mechanical properties (maximum load, stiffness and energy absorption capacity), and micro-architecture of the lumbar vertebra in the OVX osteoporotic rabbit model following treatment for 120 days. In conclusion, it was demonstrated that OsteoKing is effective in the prevention of estrogen deficiency-associated bone loss and may be a promising drug for the treatment of post-menopausal osteoporosis.
Asunto(s)
Conservadores de la Densidad Ósea/farmacología , Densidad Ósea/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Vértebras Lumbares/efectos de los fármacos , Osteoporosis/prevención & control , Absorciometría de Fotón , Fosfatasa Ácida/genética , Fosfatasa Ácida/metabolismo , Administración Oral , Animales , Estriol/análogos & derivados , Estriol/farmacología , Estrógenos/farmacología , Femenino , Isoenzimas/genética , Isoenzimas/metabolismo , Vértebras Lumbares/metabolismo , Vértebras Lumbares/patología , Osteocalcina/genética , Osteocalcina/metabolismo , Osteoporosis/etiología , Osteoporosis/genética , Osteoporosis/patología , Ovariectomía/efectos adversos , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/metabolismo , Fósforo/metabolismo , Procolágeno/genética , Procolágeno/metabolismo , Quinestrol/análogos & derivados , Conejos , Fosfatasa Ácida Tartratorresistente , Tomografía Computarizada de EmisiónRESUMEN
Nilestriol (NIL) has been applied to treat menopausal dysfunctions, yet its mechanism has remained unknown. To understand the relationship between the changes in homeostatic metabolites and ovarian oestrogen deficiency syndromes after NIL treatment, proton Nuclear Magnetic Resonance ((1)H NMR)-based metabonomic technologies were used to analyse a rat model of oestrogen deficiency. An orthogonal partial least-squares regression (OPLS) differentiation model was used on 12-week metabolic analyses of ovariectomised (OVX) rats treated or mock treated with NIL. Furthermore, data analysis using Chenomx software quantified results to identify the most significantly altered metabolite concentrations, allowing for metabolic explanations of the effects of NIL therapies. In this study, PLS results revealed that there are considerably distinct differences between treatment groups. Additionally, a total of 45 metabolites shown to have a high variation between groups were selected for target quantification. Using a one-way LSD ANOVA analysis, 32 metabolite concentrations were significantly altered in the OVX group. A total of 21 metabolites were altered significantly in the NIL-treatment group but later returned to normal. According to the OPLS VIP calculation, the metabolites most affected by NIL treatment were mostly involved in insulin resistance. In addition, abnormal concentration changes in lactate in the NIL-treatment group and 3-indoxylsulfate in the OVX group were observed. To our knowledge, this study is the first to address the molecular mechanism of NIL from a metabonomic perspective, and, more specifically, to establish a catalogue of endo-molecular changes effected by NIL in the regulation of oestrogen deficiency disorder.
Asunto(s)
Estriol/análogos & derivados , Menopausia/efectos de los fármacos , Metaboloma/fisiología , Animales , Peso Corporal , Estriol/farmacología , Femenino , Indicán/análisis , Resistencia a la Insulina , Ácido Láctico/análisis , Metabolómica , Resonancia Magnética Nuclear Biomolecular , Ovariectomía , Quinestrol/análogos & derivados , Ratas , Ratas Sprague-Dawley , Suero/química , Orina/químicaRESUMEN
Fertility control has been identified by studies in the laboratory and the field as a more appropriate and long-term control strategy for rodent pests than lethal control. In this study, we investigated the effects of quinestrol on mass of reproductive organs and on social behaviors in female and male Indochinese forest rat (Rattus andamanensis (Blyth, 1860)] using approach of morphometrics and genetics). [corrected]. A total of 16 adult females and 16 adult males were randomly assigned to 4 groups. One male and one female group were fed rice with 0.005% quinestrol by weight for 7 days, and another 2 groups were fed rice only. After 7 days, rats were assigned to 10 min dyadic encounters between groups, and investigation, aggression, defense and attack latency were quantified. All animals were killed on day 10, and reproductive organs were dissected and weighed. Dyadic encounter data showed that there were obvious changes in social behaviors of quinestrol-treated rats. Quinestrol significantly inhibited the investigative behavior of quinestrol-treated males toward control females in Rattus nitidus, but seldom affected investigation between control males and quinestrol-treated females. Aggression of control females toward quinestrol-treated males was higher than that of quinestrol-treated females, and defense of quinestrol-treated males toward control females was more remarkable than that of control males. Quinestrol remarkably decreased wet masses of epididymis and spermotophore in males and ovaries in females, but had no effect on wet masses of testes and uteri after quinestrol treatment. These results indicate that the anti-fertility effects of quinestrol on R. nitidus are attributed to not only suppressing reproductive organs but also impacting social behaviors associated with territory defense and mate choice.
Asunto(s)
Conducta Animal/efectos de los fármacos , Estriol/análogos & derivados , Fertilidad/efectos de los fármacos , Ovario/efectos de los fármacos , Ratas/fisiología , Conducta Social , Testículo/efectos de los fármacos , Agresión/efectos de los fármacos , Agresión/fisiología , Animales , Conducta Animal/fisiología , Epidídimo/efectos de los fármacos , Epidídimo/fisiología , Estriol/farmacología , Femenino , Masculino , Tamaño de los Órganos/efectos de los fármacos , Tamaño de los Órganos/fisiología , Ovario/fisiología , Testículo/fisiología , Útero/efectos de los fármacos , Útero/fisiologíaRESUMEN
Soluble adenylyl cyclase (sAC) has been hypothesized to play a role in insulin secretion. The present study aimed to investigate the interaction between adenosine 3',5'cyclic monophosphate (cAMP), volumeregulated anion channels (VRACs) and the electrogenic sodium bicarbonate (Na+HCO3) cotransporter, NBCe1, in the regulation of nutrient and hypotonicityinduced insulin release from rat pancreatic islets and tumoral insulinproducing BRINBD11 cells. In the islets, 5nitro2(3phenylpropylamino)benzoic acid (NPPB) and 5chloro2hydroxy3(thiophene2carbonyl)indole1carboxamide (tenidap) reduced glucosestimulated insulin release, however, only NPPB suppressed the enhancing action of cAMP analogs upon such a release. Insulin output from the BRINBD11 cells was stimulated by 2ketoisocaproate (KIC) or extracellular hypoosmolarity. cAMP analogs and 3isobutyl1methylxanthine increased the insulin output recorded in the isotonic medium to a greater relative extent than that in the hypotonic medium. The secretory response to KIC or hypotonicity was inhibited by NPPB or tenidap, which both also opposed the enhancing action of cAMP analogs. Inhibitors of mitogenactivated protein (MAP) kinase decreased insulin output in isotonic and hypotonic media. The inhibitor of sAC, 2hydroxyestriol, caused only a modest inhibition of insulin release, whether in the isotonic or hypotonic medium, even when tested at a concentration of 100 µM. The omission of NaHCO3 markedly decreased the secretory response to KIC or extracellular hypotonicity. The omission of Na+ suppressed the secretory response to extracellular hypotonicity. The observations of the present study do not support the hypothesis of a major role for sAC in the regulation of insulin release.
Asunto(s)
AMP Cíclico/metabolismo , Soluciones Hipotónicas/farmacología , Insulina/biosíntesis , Insulina/metabolismo , Canales Iónicos/metabolismo , Islotes Pancreáticos/metabolismo , Simportadores de Sodio-Bicarbonato/metabolismo , Animales , Aniones , Línea Celular Tumoral , AMP Cíclico/análogos & derivados , Estriol/análogos & derivados , Estriol/farmacología , Alimentos , Glucosa/farmacología , Indoles/farmacología , Secreción de Insulina , Islotes Pancreáticos/efectos de los fármacos , Soluciones Isotónicas/farmacología , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Nitrobenzoatos/farmacología , Oxindoles , Inhibidores de Fosfodiesterasa/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Ratas , Estándares de Referencia , Sodio/metabolismoRESUMEN
The glucuronidation of estriol, 16-epiestriol, and 17-epiestriol by the human UDP-glucuronosyltransferases (UGTs) of subfamilies 1A, 2A, and 2B was examined. UGT1A10 is highly active in the conjugation of the 3-OH in all these estriols, whereas UGT2B7 is the most active UGT toward one of the ring D hydroxyls, the 16-OH in estriol and 16-epiestriol, but the 17-OH in 17-epiestriol. Kinetic analyses indicated that the 17-OH configuration plays a major role in the affinity of UGT2B7 for estrogens. The glucuronidation of the different estriols by the human liver and intestine microsomes reflects the activity of UGT1A10 and UGT2B7 in combination with the tissues' difference in UGT1A10 expression. The UGT1A10 mutant 1A10-F93G exhibited much higher V(max) values than UGT1A10 in estriol and 17-epiestriol glucuronidation, but a significantly lower value in 16-epiestriol glucuronidation. To this study on estriol glucuronidation we have added experiments with 13-epiestradiol, a synthetic estradiol in which the spatial arrangement of the methyl on C18 and the hydroxyl on C17 is significantly different than in other estrogens. In comparison with estradiol glucuronidation, the C13 configuration change decreases the turnover of UGTs that conjugate the 3-OH, but increases it in UGTs that primarily conjugate the 17-OH. Unexpectedly, UGT2B17 exhibited similar conjugation rates of both the 17-OH and 3-OH of 13-espiestradiol. The combined results reveal the strong preference of UGT1A10 for the 3-OH of physiologic estrogens and the equivalently strong preference of UGT2B7 and UGT2B17 for the hydroxyls on ring D of such steroid hormones.
Asunto(s)
Estradiol/metabolismo , Estriol/análogos & derivados , Glucuronosiltransferasa/metabolismo , Biotransformación , Estradiol/análogos & derivados , Estradiol/química , Estriol/química , Estriol/metabolismo , Glucuronosiltransferasa/genética , Humanos , Hidroxilación , Isoenzimas , Cinética , Antígenos de Histocompatibilidad Menor , Modelos Moleculares , Estructura Molecular , Mutación , Proteínas Recombinantes/metabolismo , Estereoisomerismo , Relación Estructura-Actividad , Especificidad por SustratoRESUMEN
The past 20 years witnessed an enormous leap in understanding of the central regulation of whole-body energy metabolism. Genetic tools have enabled identification of the region-specific expression of peripheral metabolic hormone receptors and have identified neuronal circuits that mediate the action of these hormones on behavior and peripheral tissue functions. One of the surprising findings of recent years is the observation that brain circuits involved in metabolism regulation remain plastic through adulthood. In this review, we discuss these findings and focus on the role of neurons and glial cells in the dynamic process of plasticity, which is fundamental to the regulation of physiological and pathological metabolic events.
Asunto(s)
Metabolismo Energético/fisiología , Estriol/análogos & derivados , Hipotálamo/fisiología , Plasticidad Neuronal/fisiología , Estriol/fisiología , HumanosRESUMEN
Prune homolog 2 (Drosophila) (PRUNE2) encodes a BCH motif-containing protein that shares homology with the Cayman ataxia-related protein Caytaxin. Caytaxin is a substrate of caspase-3 and is specifically expressed at the presynapse of vesicular-type glutamate transporter (VGLUT)-positive neurons, where it plays a role in glutamate neurotransmission primarily in the cerebellum and hippocampus. Here, we showed that a novel Prune2 isoform contains a BCH motif and localizes predominantly to the synaptic cytosol, similar to Caytaxin. However, the isoform is expressed predominantly in the olfactory bulb and layer Ia of the piriform cortex, where Caytaxin is scarcely expressed. The isoform expression is upregulated during development, similar to that in the presynaptic-localizing proteins Synapsin I and Bassoon. Prune2 and its previously identified isoforms have been shown to be a susceptibility gene for Alzheimer's disease, a biomarker for leiomyosarcomas, a proapoptotic protein, and an antagonist of cellular transformation. In addition, a novel isoform may develop new roles for Prune2 at the synapse in olfactory systems.
Asunto(s)
Proteínas de Neoplasias/genética , Bulbo Olfatorio/fisiología , Vías Olfatorias/fisiología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Citosol/metabolismo , Estriol/análogos & derivados , Estriol/metabolismo , Exones/genética , Células HEK293 , Humanos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Datos de Secuencia Molecular , Proteínas de Neoplasias/química , Proteínas de Neoplasias/metabolismo , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Estructura Terciaria de Proteína/fisiologíaRESUMEN
Genistein, a major phytoestrogen of soy, is considered a potential drug for the prevention and treatment of post-menopausal osteoporosis. Mounting evidence suggested a positive correlation between genistein consumption and bone health both in vivo and in vitro. Earlier studies have revealed that genistein acted as a natural estrogen analogue which activated estrogen receptor and exerted anti-osteoporotic effect. However, it remains unclear whether PTH, the most crucial hormone that regulates mineral homeostasis, participates in the process of genistein-mediated bone protection. In the present study, we compared the therapeutic effects between genistein and nilestriol and investigated whether PTH and its specific receptor PTHR1 altered in response to genistein-containing diet in the animal model of ovariectomy. Our results showed that genistein administration significantly improved femoral mechanical properties and alleviates femoral turnover. Genistein at all doses (4.5 mg/kg, 9.0 mg/kg and 18.0 mg/kg per day, respectively) exerted improved bending strength and b-ALP limiting effects than nilestriol in the present study. However, genistein administration did not exert superior effects on bone protection than nilestriol. We also observed circulating PTH restoration in ovariectomized rats receiving genistein at the dose of 18 mg/kg per day. Meanwhile, PTHR1 abnormalities were attenuated in the presence of genistein as confirmed by RT-PCR, Western blot and immunohistochemistry. These findings strongly support the idea that besides serving as an estrogen, genistein could interact with PTH/PTHR1, causing a superior mineral restoring effect than nilestriol on certain circumstance. In conclusion, our study reported for the first time that the anti-osteoporotic effect of genistein is partly PTH/PTHR1-dependent. Genistein might be a potential option in the prevention and treatment of post-menopausal osteoporosis with good tolerance, more clinical benefits and few undesirable side effects.
Asunto(s)
Fémur/efectos de los fármacos , Genisteína/farmacología , Hormona Paratiroidea/metabolismo , Fitoestrógenos/farmacología , Sustancias Protectoras/farmacología , Receptor de Hormona Paratiroídea Tipo 1/metabolismo , Fosfatasa Alcalina/sangre , Animales , Densidad Ósea/efectos de los fármacos , Creatinina/sangre , Creatinina/orina , Modelos Animales de Enfermedad , Estriol/análogos & derivados , Estriol/química , Estriol/farmacología , Femenino , Fémur/fisiología , Genisteína/química , Genisteína/uso terapéutico , Humanos , Riñón/metabolismo , Riñón/patología , Osteoporosis Posmenopáusica/metabolismo , Osteoporosis Posmenopáusica/patología , Osteoporosis Posmenopáusica/prevención & control , Ovariectomía , Hormona Paratiroidea/sangre , Hormona Paratiroidea/orina , Fitoestrógenos/química , Fitoestrógenos/uso terapéutico , Sustancias Protectoras/química , Sustancias Protectoras/uso terapéutico , Quinestrol/análogos & derivados , Ratas , Ratas Sprague-Dawley , Receptor de Hormona Paratiroídea Tipo 1/genética , Resistencia a la TracciónRESUMEN
OBJECTIVE: To investigate the influence of nilestriol (CCE3) and exercise on bone size and bone mass in ovariectomized (OVX) rats. METHODS: Forty eight (48) female Sprague-Dawley (SD) rats were divided randomly into six groups: (1) Normal control group, (2) Sham OVX group, (3) OVX group, (4) OVX + CCE3 group, (5)) OVX + exercise group, (6) OVX + CCE3 + exercise group. CCE (0.5 mg/kg per week) was given to the rats in OVX + CCE3 group and OVX + CCE3 + exercise group from the 2nd day after the operation for 11 weeks. Exercise training was loaded to the rats in exercise groups from the 7th day after operation, each rat was subjected to running on 0 dig angle runway with the speed of 16 m per minute, 45 minutes per day, 5 days per week for 10 weeks. RESULTS: The bone diameter, bone volume, bone wet weight, bone dry weight, bone ashes weight in the OVX group were lower than those in the sham OVX group (P < 0.01). All of these measurements were increased in OVX + CCE3 group and OVX + exercise group when compared with the OVX group, but were still lower than those in sham OVX group (P < 0.01). Exercise enhances the effect of CCE3 (P < 0.01). CONCLUSION: Exercise enhances the effects of CCE3 on the improvement of both bone size and quantity in OVX rats.
Asunto(s)
Huesos/anatomía & histología , Estriol/análogos & derivados , Osteoporosis/terapia , Esfuerzo Físico , Animales , Densidad Ósea , Terapia Combinada , Estriol/administración & dosificación , Femenino , Osteoporosis/etiología , Ovariectomía , Quinestrol/análogos & derivados , Distribución Aleatoria , Ratas , Ratas Sprague-DawleyRESUMEN
In anticancer research where the focus is on finding agents that induces cell death while leaving non-tumorigenic cells less affected, a novel 2-methoxyestradiol derivative has come forth. 2-Methoxyestradiol-bis-sulfamate (2-MeOE2bisMATE) is a 2-methoxyestradiol derivative produced by bis-sulphamoylation, which possesses increased antiproliferative activity and biological availability. Several questions remain regarding the type of cell death mechanisms and possible induction of autophagy by 2-MeOE2bisMATE. The aim of this in vitro study was to investigate the cell death mechanisms exerted by 2-MeOE2bisMATE in an adenocarcinoma cell line (MCF-7) by analyzing its influence on cell growth, morphology, and possible induction of cell death. Spectrophotometry (crystal violet staining), transmission electron microscopy (TEM), light microscopy (hematoxylin and eosin staining), and fluorescent microscopy (Hoechst 33342, propidium iodide and acridine orange) were employed. Spectrophotometrical studies indicated that 2-MeOE2bisMATE decreased cell numbers to 75% in MCF-7 cells after 24 h and to 47% after 48 h of exposure. TEM demonstrated membrane blebbing, nuclear fragmentation, and chromatin condensation indicating the hallmarks of apoptosis. Light microscopy revealed the presence of several cells blocked in metaphase, and apoptotic cells were also observed. Fluorescent microscopy demonstrated increased lysosomal staining; suggesting the induction of autophagy. 2-MeOE2bisMATE shows therapeutic potential, as an, anticancer agent, and the investigation of the cell death mechanisms used by 2-MeOE2bisMATE, thus, warrants further investigation.