RESUMEN
Administration of Chemotherapeutics, especially doxorubicin (DOX) and cyclophosphamide (CPS), is commonly associated with adverse effects such as myelosuppression and cardiotoxicity. At this time, few approved therapeutic options are currently available for the management of chemotherapy-associated cardiotoxicity. Thus, identification of novel therapeutics with potent cardioprotective properties and minimal adverse effects are pertinent in treating Doxorubicin and Cyclophosphamide-induced cardiotoxicity. Oroxylum indicum extract (OIE, Sabroxy®) is a natural product known to possess several beneficial biological functions including antioxidant, anti-inflammatory and cytoprotective effects. We therefore set to investigate the cardioprotective effects of OIE against Doxorubicin and Cyclophosphamide-induced cardiotoxicity and explore the potential cardioprotective mechanisms involved. Adult male mice were treated with DOX and CPS in combination, OIE alone, or a combination of OIE and DOX & CPS. Swimming test was performed to assess cardiac function. Markers of oxidative stress were assessed by levels of reactive oxygen species (ROS), nitrite, hydrogen peroxide, catalase, and glutathione content. The activity of interleukin converting enzyme and cyclooxygenase was determined as markers of inflammation. Mitochondrial function was assessed by measuring Complex-I activity. Apoptosis was assessed by Caspase-3 and protease activity. Mice treated with DOX and CPS exhibited reduced swim rate, increased oxidative stress, increased inflammation, and apoptosis in the heart tissue. These cardiotoxic effects were significantly reduced by co-administration of OIE. Furthermore, computational molecular docking studies revealed potential binding of DOX and CPS to tyrosine hydroxylase which validated our in vivo findings regarding the inhibition of tyrosine hydroxylase activity. Our current findings indicated that OIE counteracts Doxorubicin and Cyclophosphamide-induced cardiotoxicity-through inhibition of ROS-mediated apoptosis and by blocking the effect on tyrosine hydroxylase. Taken together, our findings suggested that OIE possesses cardioprotective effects to counteract potentially fatal cardiac complications associated with chemotherapy treatment.
Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/farmacología , Bignoniaceae , Cardiopatías/prevención & control , Mitocondrias Cardíacas/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Antiinflamatorios/aislamiento & purificación , Antioxidantes/aislamiento & purificación , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/metabolismo , Bignoniaceae/química , Cardiotoxicidad , Ciclofosfamida , Modelos Animales de Enfermedad , Doxorrubicina , Cardiopatías/inducido químicamente , Cardiopatías/metabolismo , Cardiopatías/patología , Mediadores de Inflamación/metabolismo , Masculino , Ratones Endogámicos C57BL , Mitocondrias Cardíacas/metabolismo , Mitocondrias Cardíacas/patología , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/aislamiento & purificación , Especies Reactivas de Oxígeno/metabolismo , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Aspartame is one of the most widely used artificial sweeteners globally. Data concerning acute neurotoxicity of aspartame is controversial, and knowledge on its chronic effect is limited. In the current study, we investigated the chronic effects of aspartame on ionic homeostasis and regional monoamine neurotransmitter concentrations in the brain. Our results showed that aspartame at high dose caused a disturbance in ionic homeostasis and induced apoptosis in the brain. We also investigated the effects of aspartame on brain regional monoamine synthesis, and the results revealed that there was a significant decrease of dopamine in corpus striatum and cerebral cortex and of serotonin in corpus striatum. Moreover, aspartame treatment significantly alters the tyrosine hydroxylase activity and amino acids levels in the brain. Our data suggest that chronic use of aspartame may affect electrolyte homeostasis and monoamine neurotransmitter synthesis dose dependently, and this might have a possible effect on cognitive functions.
Asunto(s)
Apoptosis , Aspartame/efectos adversos , Corteza Cerebral/metabolismo , Cuerpo Estriado/metabolismo , Antagonistas de Dopamina/efectos adversos , Edulcorantes no Nutritivos/efectos adversos , Antagonistas de la Serotonina/efectos adversos , Animales , Aspartame/administración & dosificación , Corteza Cerebral/enzimología , Cuerpo Estriado/enzimología , Antagonistas de Dopamina/administración & dosificación , Masculino , Proteínas del Tejido Nervioso/antagonistas & inhibidores , Proteínas del Tejido Nervioso/metabolismo , Neuronas/enzimología , Neuronas/metabolismo , Síndromes de Neurotoxicidad/enzimología , Síndromes de Neurotoxicidad/etiología , Síndromes de Neurotoxicidad/metabolismo , Edulcorantes no Nutritivos/administración & dosificación , Fenilalanina/agonistas , Fenilalanina/metabolismo , Distribución Aleatoria , Ratas Wistar , Antagonistas de la Serotonina/administración & dosificación , ATPasa Intercambiadora de Sodio-Potasio/antagonistas & inhibidores , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Pruebas de Toxicidad Crónica , Triptófano/antagonistas & inhibidores , Triptófano/metabolismo , Tirosina/agonistas , Tirosina/metabolismo , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Tirosina 3-Monooxigenasa/metabolismo , Desequilibrio Hidroelectrolítico/enzimología , Desequilibrio Hidroelectrolítico/etiología , Desequilibrio Hidroelectrolítico/metabolismoRESUMEN
This study was aimed to test our hypothesis that the developing brain operates as an endocrine organ before the establishment of the blood-brain barrier (BBB), in rats up to the first postnatal week. Dopamine (DA) was selected as a marker of the brain endocrine activity. The hypothesis was supported by the observations in rats of: (i) the physiological concentration of DA in peripheral blood of fetuses and neonates, before the BBB establishment, and its drop by prepubertal period, after the BBB development; (ii) a drop of the DA concentration in the brain for 54% and in blood for 74% on the 3rd postnatal day after the intraventricular administration of 50 µg of α-methyl-p-tyrosine, an inhibitor of DA synthesis, with no changes in the DA metabolism in peripheral DA-producing organs. Thus, the developing brain is a principal source of circulating DA which is capable of providing an endocrine regulation of peripheral organs and the brain.
Asunto(s)
Encéfalo/crecimiento & desarrollo , Dopamina/metabolismo , Sistemas Neurosecretores/crecimiento & desarrollo , Ácido 3,4-Dihidroxifenilacético/sangre , Ácido 3,4-Dihidroxifenilacético/metabolismo , Animales , Barrera Hematoencefálica/embriología , Barrera Hematoencefálica/crecimiento & desarrollo , Encéfalo/embriología , Encéfalo/metabolismo , Dopamina/sangre , Femenino , Hipotálamo/metabolismo , Masculino , Mesencéfalo/metabolismo , Sistemas Neurosecretores/embriología , Embarazo , Ratas , Ratas Wistar , Rombencéfalo/metabolismo , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , alfa-Metiltirosina/farmacologíaRESUMEN
It has been reported that neuropeptide Y (NPY) contributes to the behavioral response of amphetamine (AMPH), a psychostimulant. The present study examined whether protein kinase C (PKC)-λ signaling was involved in this action. Moreover, possible roles of glutathione peroxidase (GP) and melanocortin receptor 4 (MC4R) were also examined. Rats were given AMPH daily for 4 days. Hypothalamic NPY, PKCλ, GP and MC4R were determined and compared. Pretreatment with α-methyl-para-tyrosine could block AMPH-induced anorexia, revealing that endogenous catecholamine was involved in regulating AMPH anorexia. PKCλ, GP and MC4R were increased with maximal response on Day 2 during AMPH treatment, which were concomitant with the decreases in NPY. cAMP response element binding protein (CREB) DNA binding activity was increased during AMPH treatment, revealing the involvement of CREB-dependent gene transcription. An interruption of cerebral PKCλ transcript could partly block AMPH-induced anorexia and partly reverse NPY, MC4R and GP mRNA levels to normal. These results suggest that PKCλ participates in regulating AMPH-induced anorexia via a modulation of hypothalamic NPY gene expression and that increases of GP and MC4R may contribute to this modulation. Our results provided molecular evidence for the regulation of AMPH-induced behavioral response.
Asunto(s)
Anfetamina/farmacología , Depresores del Apetito/farmacología , Estimulantes del Sistema Nervioso Central/farmacología , Glutatión Peroxidasa/metabolismo , Hipotálamo/efectos de los fármacos , Isoenzimas/deficiencia , Neuropéptido Y/metabolismo , Proteína Quinasa C/deficiencia , Receptor de Melanocortina Tipo 4/metabolismo , Animales , Anorexia/inducido químicamente , Anorexia/enzimología , Anorexia/genética , Elementos sin Sentido (Genética)/administración & dosificación , Conducta Animal/efectos de los fármacos , Sitios de Unión , Catecolaminas/metabolismo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Relación Dosis-Respuesta a Droga , Ingestión de Alimentos/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Regulación de la Expresión Génica , Técnicas de Silenciamiento del Gen , Glutatión Peroxidasa/genética , Hipotálamo/enzimología , Inyecciones Intraventriculares , Isoenzimas/genética , Masculino , Neuropéptido Y/genética , Proteína Quinasa C/genética , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Receptor de Melanocortina Tipo 4/genética , Factores de Tiempo , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Tirosina 3-Monooxigenasa/metabolismo , alfa-Metiltirosina/farmacologíaRESUMEN
In this study, the effect of pharmacological inhibition of catecholaminergic activity on hCG-induced spawning was evaluated and correlated with tyrosine hydroxylase (TH) activity, the rate-limiting enzyme in catecholamine biosynthesis. Gravid female H. fossilis collected in both prespawning and spawning phases were given alpha-methylparatyrosine (alpha-MPT: 250 microg/g body weight, ip, an irreversible inhibitor of TH) and human chorionic gonadotropin (hCG: 100 IU/fish, ip) alone or in combination. The fish were sampled at different intervals for measuring hypothalamic and ovarian TH activity and checking spawning response. The administration of hCG resulted in ovulation and spawning in both phases with a higher response in the spawning phase. The administration of alpha-MPT did not induce any response, like the control fish. In the hCG + alpha-MPT groups, the spawning response of hCG was significantly inhibited and delayed by the inhibitor. The spawning response of hCG was accompanied by a significant increase in both hypothalamic and ovarian TH activity at 6 and 12 h of the injection. However, at 24hr the activity decreased except in the spawning phase. The alpha-MPT treatment inhibited TH activity significantly in a duration-dependent manner. In the hCG + alpha-MPT groups, enzyme activity was inhibited at all duration. The results indicate the involvement of catecholamines during the hCG-induced spawning and the specific functional nature of the involvement needs further investigation.
Asunto(s)
Gonadotropina Coriónica/farmacología , Ovulación/efectos de los fármacos , Reproducción/efectos de los fármacos , Conducta Sexual Animal/efectos de los fármacos , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , alfa-Metiltirosina/farmacología , Animales , Catecolaminas/metabolismo , Bagres , Inhibidores Enzimáticos/farmacología , Femenino , Humanos , Hipotálamo/efectos de los fármacos , Sustancias para el Control de la Reproducción/farmacologíaRESUMEN
There is no effective treatment for cocaine addiction despite extensive knowledge of the neurobiology of drug addiction. Here we show that a selective aldehyde dehydrogenase-2 (ALDH-2) inhibitor, ALDH2i, suppresses cocaine self-administration in rats and prevents cocaine- or cue-induced reinstatement in a rat model of cocaine relapse-like behavior. We also identify a molecular mechanism by which ALDH-2 inhibition reduces cocaine-seeking behavior: increases in tetrahydropapaveroline (THP) formation due to inhibition of ALDH-2 decrease cocaine-stimulated dopamine production and release in vitro and in vivo. Cocaine increases extracellular dopamine concentration, which activates dopamine D2 autoreceptors to stimulate cAMP-dependent protein kinase A (PKA) and protein kinase C (PKC) in primary ventral tegmental area (VTA) neurons. PKA and PKC phosphorylate and activate tyrosine hydroxylase, further increasing dopamine synthesis in a positive-feedback loop. Monoamine oxidase converts dopamine to 3,4-dihydroxyphenylacetaldehyde (DOPAL), a substrate for ALDH-2. Inhibition of ALDH-2 enables DOPAL to condense with dopamine to form THP in VTA neurons. THP selectively inhibits phosphorylated (activated) tyrosine hydroxylase to reduce dopamine production via negative-feedback signaling. Reducing cocaine- and craving-associated increases in dopamine release seems to account for the effectiveness of ALDH2i in suppressing cocaine-seeking behavior. Selective inhibition of ALDH-2 may have therapeutic potential for treating human cocaine addiction and preventing relapse.
Asunto(s)
Aldehído Deshidrogenasa/antagonistas & inhibidores , Aldehído Deshidrogenasa/uso terapéutico , Alcaloides de Berberina/metabolismo , Trastornos Relacionados con Cocaína/prevención & control , Antagonistas de Dopamina/metabolismo , Proteínas Mitocondriales/antagonistas & inhibidores , Proteínas Mitocondriales/uso terapéutico , Aldehído Deshidrogenasa Mitocondrial , Animales , Cocaína/administración & dosificación , Señales (Psicología) , Modelos Animales de Enfermedad , Dopamina/biosíntesis , Activación Enzimática , Infusiones Intravenosas , Ratas , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Estrogens modulate critical homeostatic functions of the hypothalamus such as temperature regulation, sexual behavior and sleep with the most pronounced effects in rats occurring during the dark-phase. The neurochemical signals underlying estrogenic regulation of these hypothalamic functions have not been clearly identified, possibly due to the fact that previous studies have not explored the effects of estrogen treatments on neuronal signaling during the dark-phase. In the present study, ovariectomized rats received estradiol benzoate (5 microg/rat for 7 days, s.c.) and norepinephrine and dopamine levels were measured in the preoptic area of the hypothalamus across the light/dark cycle using in vivo microdialysis. Estradiol benzoate treatment increased extracellular norepinephrine and dopamine levels relative to vehicle treatment during the dark-phase. Increases in norepinephrine and dopamine were first detected by 30 min and 5.5h after lights-off, respectively. Subsequent increases in norepinephrine and dopamine were also noted throughout the 9.5-h collection period. The effect of estradiol benzoate on catecholamine release did not correlate with increases in either tyrosine hydroxylase (TH) protein expression or activity levels in the anterior hypothalamus, although a marked decrease in TH activity correlated with a rise in extracellular norepinephrine at the beginning of the dark-phase. We conclude that subchronic estradiol benzoate treatment increases extracellular catecholamine levels in the preoptic area of the hypothalamus during the dark-phase without a concomitant increase in neurotransmitter biosynthesis. The estradiol benzoate-induced increases in norepinephrine and dopamine levels in the preoptic area during the dark-phase may play an important role in modulating critical hypothalamic functions.
Asunto(s)
Catecolaminas/metabolismo , Oscuridad , Estradiol/análogos & derivados , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Ovariectomía , Animales , Dopamina/metabolismo , Estradiol/farmacología , Espacio Extracelular/efectos de los fármacos , Espacio Extracelular/metabolismo , Femenino , Hipotálamo/citología , Microdiálisis , Norepinefrina/metabolismo , Área Preóptica/citología , Área Preóptica/efectos de los fármacos , Área Preóptica/metabolismo , Ratas , Ratas Sprague-Dawley , Factores de Tiempo , Tirosina 3-Monooxigenasa/antagonistas & inhibidoresRESUMEN
Striatal administration of 1-methyl-4-phenylpyridinium (MPP(+)), the active metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), causes nigrostriatal dopaminergic pathway damage similar to that observed in Parkinson's disease. Copper acts as a prosthetic group of several antioxidant enzymes and recent data show that copper attenuated MPP(+)-evoked neurotoxicity. We evaluated the effect of copper (as a supplement) upon proteins nitration (60 kDa) and tyrosine hydroxylase (TH) inactivation induced by MPP(+) (10 microg/8 microL) injection into the rat striatum. Copper pretreatment (10 micromol/kg i.p.) prevented both MPP(+)-induced proteins nitration and TH inactivation. Copper treatment also prevented the dopamine-depleting effect of MPP(+) injection. Those results were accompanied by a significant reduction of enzymatic activity of the constitutive nitric oxide synthase (cNOS), whereas, the protein levels of the three isoforms of NOS remained unchanged. Results indicate that the effect of copper against MPP(+)-induced proteins nitration and TH inactivation in the striatum of rat may be mediated by a reduction of cNOS activity.
Asunto(s)
Sulfato de Cobre/farmacología , Intoxicación por MPTP/prevención & control , Neostriado/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Nitratos/metabolismo , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina , Animales , Western Blotting , Dopamina/metabolismo , Dopaminérgicos/toxicidad , Electroforesis en Gel de Poliacrilamida , Masculino , Neostriado/efectos de los fármacos , Óxido Nítrico Sintasa de Tipo I/biosíntesis , Enfermedad de Parkinson Secundaria/prevención & control , Ratas , Ratas WistarRESUMEN
Phenylpropanolamine (PPA) is an appetite suppressant. Repeated treatment with PPA can decrease food intake on initial days, but on subsequent days, food intake gradually returns to normal (tolerant effect). In an attempt to investigate the underlying mechanisms of PPA tolerance, the authors examined the roles of catecholamine (CAT) and hypothalamic neuropeptide Y (NPY) genome. Results revealed that pretreatment with either bupropion, a CAT transporter inhibitor, or a-methylparatyrosine, a tyrosine hydroxylase inhibitor, modulated the effect of PPA tolerance. Moreover, results also revealed that the alteration in NPY messenger RNA level coincided with the change of feeding behavior during PPA treatment and that infusions of NPY antisense oligonucleotide into the cerebroventricle abolished the effect of PPA tolerance. These findings suggest that cerebral CAT and hypothalamic NPY genome are involved in the development of tolerance to PPA-induced appetite suppression.
Asunto(s)
Depresores del Apetito/farmacología , Catecolaminas/genética , Catecolaminas/fisiología , Sistema Nervioso Central/metabolismo , Hipotálamo/metabolismo , Neuropéptido Y/genética , Neuropéptido Y/fisiología , Fenilpropanolamina/farmacología , Anfetamina/farmacología , Animales , Peso Corporal/efectos de los fármacos , Bupropión/farmacología , Cateterismo , Sistema Nervioso Central/efectos de los fármacos , Inhibidores de Captación de Dopamina/farmacología , Tolerancia a Medicamentos , Electroforesis en Gel de Poliacrilamida , Conducta Alimentaria/efectos de los fármacos , Hipotálamo/efectos de los fármacos , Inyecciones Intraventriculares , Masculino , Oligonucleótidos Antisentido/farmacología , ARN Mensajero/biosíntesis , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiología , Tirosina 3-Monooxigenasa/antagonistas & inhibidoresRESUMEN
BACKGROUND AND PURPOSE: Methylamine is an endogenous aliphatic amine exhibiting anorexigenic properties in mice. The aim of this work was to show whether methylamine also modifies feeding behaviour in rats and, if so, to identify the mediator(s) responsible for such effects. EXPERIMENTAL APPROACH: Microdialysis experiments with the probe inserted in the periventricular hypothalamic nucleus were carried out in 12 h starved, freely moving rats. Collected perfusate samples following methylamine injection (i.c.v.) were analysed for nitric oxide by chemiluminescence and for dopamine and 5-hydroxytryptamine content by HPLC. Kv1.6 potassium channel expression was reduced by antisense strategy and this decrease quantified by semi-quantitative RT-PCR analysis. KEY RESULTS: Methylamine showed biphasic dose-related effects on rat feeding. At doses of 15-30 microg per rat, it was hyperphagic whereas higher doses (60-80 microg) were hypophagic. Methylamine stimulated central nitric oxide (+115% vs. basal) following hyperphagic and dopamine release (60% over basal values) at hypophagic doses, respectively. Treatment with L-N(G)-nitro-L-arginine-methyl ester (i.c.v. 2 microg 10 microl(-1)) or with alpha-methyl-p-tyrosine (i.p. 100 mg kg(-1)) before methylamine injection, reduced nitric oxide output and hyperphagia, or dopamine release and hypophagia respectively. Moreover, hypophagia and hyperphagia, as well as nitric oxide and dopamine release were significantly reduced by down-regulating brain Kv1.6 potassium channel expression. CONCLUSIONS AND IMPLICATIONS: The effects of methylamine on feeding depend on the hypothalamic release of nitric oxide and dopamine as a result of interaction at the Kv1.6 channels. The study of methylamine levels in the CNS may provide new perspectives on the physiopathology of alimentary behaviour.
Asunto(s)
Depresores del Apetito/metabolismo , Regulación del Apetito , Dopamina/metabolismo , Ayuno , Hipotálamo/metabolismo , Metilaminas/metabolismo , Óxido Nítrico/metabolismo , Animales , Depresores del Apetito/administración & dosificación , Regulación del Apetito/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Hiperfagia/metabolismo , Hiperfagia/fisiopatología , Hipotálamo/efectos de los fármacos , Inyecciones Intraventriculares , Canal de Potasio Kv1.6/efectos de los fármacos , Canal de Potasio Kv1.6/genética , Canal de Potasio Kv1.6/metabolismo , Masculino , Metilaminas/administración & dosificación , Microdiálisis , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa/metabolismo , Oligonucleótidos Antisentido/genética , Oligonucleótidos Antisentido/metabolismo , Ratas , Ratas Wistar , Serotonina/metabolismo , Factores de Tiempo , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Tirosina 3-Monooxigenasa/metabolismo , alfa-Metiltirosina/farmacologíaRESUMEN
BACKGROUND: The ability of animals to make morphine has been in question for the last 30 years. Studies have demonstrated that animals do contain morphine precursors and metabolites, as well as the ability to use some morphine precursors to make morphine. MATERIAL/METHODS: The present study uses excised ganglia from the marine invertebrate Mytilus edulis as well as whole animals. Morphine and dopamine levels were determined by high performance liquid chromatography coupled to electrochemical detection and radioimmunoassay. Tissues and whole animals were also exposed to morphine precursors and exposed to the CYP2D6 inhibitor quinidine and the tyrosine hydroxylase inhibitor alpha-methyl-para-tyrosine (AMPT). Additionally, via RT-PCR, a cDNA fragment of the CYP2D6 enzyme in the ganglia of M. edulis was identified. RESULTS: Pedal ganglia incubated with either tyramine or tyrosine, or whole animals receiving injections, exhibited a statistically significant concentration- and time-dependent increase in their endogenous morphine and dopamine levels (2.51 +/- 0.76 ng/g for tyrosine and 2.39 +/- 0.64 ng/g for tyramine compared to approximately 1.0 ng/g morphine wet weight). Incubation with quinidine and/or AMPT diminished ganglionic morphine and dopamine synthesis at various steps in the synthesis process. We also demonstrated that CYP2D6 mediates the tyramine to dopamine step in this process, as did tyrosine hydroxylase in the step from tyrosine to L-DOPA. Furthermore, via RT-PCR, we identified a cDNA fragment of the CYP2D6 enzyme in the ganglia, which exhibits 94% sequence identity with its human counterpart. Evidence that tyrosine and tyramine were, in part, being converted to dopamine then morphine, and that this process can be inhibited by altering either or both CYP2D6 or tyrosine hydroxylase, is also provided. CONCLUSIONS: It appears that animals have the ability to make morphine. This process also appears to be dynamic in that the inhibition of one pathway allows the other to continue with morphine synthesis. Moreover, dopamine and morphine synthesis were coupled.
Asunto(s)
Dopamina/metabolismo , Ganglios de Invertebrados/efectos de los fármacos , Morfina/metabolismo , Tiramina/farmacología , Tirosina/farmacología , Animales , Secuencia de Bases , Citocromo P-450 CYP2D6/genética , Inhibidores del Citocromo P-450 CYP2D6 , ADN Complementario/genética , Inhibidores Enzimáticos/farmacología , Ganglios de Invertebrados/enzimología , Ganglios de Invertebrados/metabolismo , Datos de Secuencia Molecular , Mytilus edulis/anatomía & histología , Quinidina/farmacología , ARN Mensajero/análisis , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Regulación hacia Arriba , alfa-Metiltirosina/farmacologíaRESUMEN
It is known that, under stress conditions the hypothalamo-pituitary-adrenal (HPA) axis is stimulated and catecholamine production is increased. Adrenomedullin (ADM) is a novel peptide that elicits a long-vasorelaxation, and participates in blood pressure regulation via different mechanisms. In the present study, we investigated the administration of ADM on tyrosine hydroxylase (TH) enzyme activity in cold exposed rats. Four groups of Sprague-Dawley rats were studied for their TH enzyme activity in the adrenal medulla and hypothalamus. In addition to measuring blood pressure in these rats, TH enzyme activity in both the adrenal medulla and hypothalamus were examined in four groups of Sprague-Dawley rats: animals exposed to room temperature and cold stress (8 masculine C, 48 h), and rats injected with ADM (1.0 nmol/kg, i.v.) alone and/or together with cold stress. TH activity was shown to be increased in cold treated groups and decreased in ADM and ADM + cold stress group. Our findings appear to suggest that external ADM application caused an opposite effect on the same system in rats, decreasing the activity of tyrosine hydroxylase (TH) enzyme activity. Furthermore, externally applied ADM was shown to produce its expected hypotensive effect in cold-stressed rats. Our results suggest that a possible explanation for the effects of ADM is that, the uptake of ADM under cold stress may effect TH activity in studied tissues.
Asunto(s)
Presión Sanguínea/efectos de los fármacos , Frío , Péptidos/farmacología , Estrés Fisiológico/fisiopatología , Tirosina 3-Monooxigenasa/metabolismo , Médula Suprarrenal/enzimología , Adrenomedulina , Animales , Hipotálamo/enzimología , Inyecciones Intravenosas , Masculino , Péptidos/administración & dosificación , Ratas , Ratas Sprague-Dawley , Estrés Fisiológico/enzimología , Tirosina 3-Monooxigenasa/antagonistas & inhibidoresRESUMEN
The aqueous extract of Mallotus japonicus (Euphorbiaceae) showed an inhibitory effect on bovine adrenal tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of catecholamine. The present study was undertaken to investigate the effects of bergenin and norbergenin, constituents of the aqueous extract of Mallotus japonicus on bovine adrenal TH. Bergenin and norbergenin inhibited the TH activity by 29.0% and 53.4% at a concentration of 20 microg/mL, respectively, and exhibited noncompetitive inhibition of TH activity with the substrate l-tyrosine. The inhibition of TH activity and the inhibitory effect of norbergenin was more potent than that of bergenin. From these results, it is presumed that bergenin and norbergenin may be the active components of Mallotus japonicus in inhibiting TH, and these inhibitory effects may be partially responsible for the clinical use of Mallotus japonicus in treating peptic ulcer by reducing the availability of dopa/dopamine in vivo.
Asunto(s)
Benzopiranos/farmacología , Medicamentos Herbarios Chinos/farmacología , Inhibidores Enzimáticos/farmacología , Mallotus (Planta) , Fitoterapia , Tirosina 3-Monooxigenasa/efectos de los fármacos , Glándulas Suprarrenales/efectos de los fármacos , Glándulas Suprarrenales/enzimología , Animales , Benzopiranos/administración & dosificación , Benzopiranos/uso terapéutico , Bovinos , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/uso terapéutico , Inhibidores Enzimáticos/administración & dosificación , Inhibidores Enzimáticos/uso terapéutico , Tirosina 3-Monooxigenasa/antagonistas & inhibidoresRESUMEN
OBJECTIVE: Uraemia often provokes various neurological disorders, such as mental changes, malperception, confusion, seizures and coma. Since changes in neurotransmissions induce neurological symptoms, we investigated changes in the monoamine metabolism and motor activity in uraemic rats. DESIGN: Prospective, randomised, controlled animal study. SUBJECTS: Male Wistar rats. INTERVENTIONS: Acute renal failure was induced by occlusion of bilateral renal arteries for 60 min, and the motor activity and brain monoamine turnover were examined 48 h later. The brain monoamine turnover was evaluated by the depletion of norepinephrine (NE) and dopamine (DA) induced by alpha-methyl-p-tyrosine (alpha-MT), or the accumulation of 5-hydroxyindoleacetic acid (5-HIAA) induced by probenecid. MEASUREMENTS AND RESULTS: Marked damage in renal function was found in animals subjected to renal ischaemia 48 h after the operation. The motor activity of the uraemic rats was impaired. The turnover of DA in the striatum, mesencephalon and hypothalamus was decreased in these rats. The turnover of NE and 5-hydroxytryptamine (5-HT) was unchanged in all regions examined. CONCLUSIONS: Suppression of the central DA turnover appears to be involved in the impairment of motor activity in uraemic rats.
Asunto(s)
Lesión Renal Aguda/metabolismo , Lesión Renal Aguda/fisiopatología , Química Encefálica , Modelos Animales de Enfermedad , Dopamina/metabolismo , Actividad Motora , Uremia/metabolismo , Uremia/fisiopatología , Lesión Renal Aguda/complicaciones , Animales , Química Encefálica/efectos de los fármacos , Coma/etiología , Confusión/etiología , Cuerpo Estriado/química , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/metabolismo , Hipotálamo/química , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Masculino , Mesencéfalo/química , Mesencéfalo/efectos de los fármacos , Mesencéfalo/metabolismo , Actividad Motora/efectos de los fármacos , Norepinefrina/metabolismo , Trastornos de la Percepción/etiología , Probenecid/farmacología , Estudios Prospectivos , Distribución Aleatoria , Ratas , Ratas Wistar , Convulsiones/etiología , Serotonina/metabolismo , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Uremia/complicaciones , Uricosúricos/farmacología , alfa-Metiltirosina/farmacologíaRESUMEN
The effect of tetrandrine, a bis-benzylisoquinoline alkaloid, on dopamine biosynthesis in PC12 cells was investigated. Tetrandrine at a concentration of 3.0 microM decreased dopamine content by 59.4% (IC50 = 2.4 microM) and intracellular tyrosine hydroxylase (TH) activity was inhibited by the treatment of tetrandrine (49.8% inhibition at 3.0 microM) compared with control. We next examined the effects of tetrandrine on the kinetics of PC12 TH. The PC12 TH was obtained from PC12 cells with minor purification. Tetrandrine inhibited the PC12 TH activity by 40.6% at a concentration of 45 microM and exhibited noncompetitive inhibition on the enzyme using L-tyrosine as a substrate (Ki = 60.8 microM). These results suggest that the inhibition of TH activity by tetrandrine may partially contribute to the decrease in dopamine biosynthesis in PC12 cells.
Asunto(s)
Alcaloides/farmacología , Bencilisoquinolinas , Antagonistas de Dopamina/farmacología , Dopamina/biosíntesis , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Animales , Células PC12 , RatasRESUMEN
The present study was undertaken to investigate the effects of ginsenosides on bovine adrenal tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. Ginsenoside-Rb1, Rc, Re and Rg, inhibited the TH activity by 51.5, 25.4, 31.3, 44.3 and 43.3%, respectively, at a concentration of 80 microg/ml. Ginsenoside-Rb1, Rc, Re and Rg1 exhibited noncompetitive inhibition of TH activity with a substrate L-tyrosine. From these results, it is presumed that the effects of ginsenosides on TH activity observed in vitro might be also produced in vivo, and thereby the inhibitory effects of ginsenosides on TH activity may be partially responsible for the antidopaminergic action of ginsenosides by reducing the availability of dopamine at the presynaptic dopamine receptor in vivo.
Asunto(s)
Glándulas Suprarrenales/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Saponinas/farmacología , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Glándulas Suprarrenales/enzimología , Animales , Bovinos , Ginsenósidos , Cinética , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Tyrosine hydroxylase (TH) activity, the rate-limiting step in the synthesis of catecholamines, was quantified in the preoptic area-hypothalamus of adult male Japanese quail by a new assay measuring the tritiated water production from 3,5-[3H]-L-tyrosine. Maximal levels of activity were observed at a 20-25 microM concentration of substrate, with more than 50% inhibition of the activity being recorded at a 100 microM concentration. TH activity was linear as a function of the incubation time during the first 20 min and maximal at a pH of 6.0. TH was heterogeneously distributed in the quail brain with highest levels of activity being found (in decreasing order) in the mesencephalon, diencephalon, and telencephalon. Given the large size of the telencephalon, this is the brain area that contains, as a whole, the highest level of enzyme activity. TH inhibitors that have been well-characterized in mammals, such as 3-iodo-L-tyrosine and L-alpha-methyl-p-tyrosine (AMPT) completely inhibited the enzyme activity at a 100 microM concentration. In mammals, the accumulation of catecholamines exerts a negative feedback control on TH activity. Similar controls were observed in the quail brain. Two inhibitors of the DOPA decarboxylase that should lead to accumulation of DOPA depressed TH activity by 60% or more, and the inhibitor of the dopamine beta-hydroxylase, fusaric acid that should cause an accumulation of dopamine, suppressed 90% of the TH activity. The addition of exogenous DOPA, dopamine, or norepinephrine to the brain homogenates also strongly inhibited TH activity, independently confirming the feedback effects of the enzyme products on the enzyme activity. These data demonstrate that TH activity in the quail brain is heterogeneously distributed and acutely regulated, as it is in mammals, by the accumulation of its products and of the derived catecholamines.
Asunto(s)
Encéfalo/enzimología , Coturnix/metabolismo , Tirosina 3-Monooxigenasa/metabolismo , Animales , Inhibidores de Descarboxilasas de Aminoácidos Aromáticos , Catecolaminas/farmacología , Dopamina beta-Hidroxilasa/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Retroalimentación , Hipotálamo/enzimología , Masculino , Área Preóptica/enzimología , Distribución Tisular/fisiología , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , alfa-Metiltirosina/farmacologíaRESUMEN
We have previously reported that atrial natriuretic factor (ANF) increased neuronal norepinephrine (NE) uptake and reduced basal and evoked neuronal NE release. Changes in NE uptake and release are generally associated to modifications in the synthesis and/or turnover of the amine. On this basis, the aim of the present work was to study ANF effects in the rat hypothalamus on the following processes: endogenous content, utilization and turn-over of NE; tyrosine hydroxylase (TH) activity; cAMP and cGMP accumulation and phosphatidylinositol hydrolysis. Results showed that centrally applied ANF (100 ng/microl/min) increased the endogenous content of NE (45%) and diminished NE utilization. Ten nM ANF reduced the turnover of NE (53%). In addition, ANF (10 nM) inhibited basal and evoked (with 25 mM KCl) TH activity (30 and 64%, respectively). Cyclic GMP levels were increased by 10 nM ANF (100%). However, neither cAMP accumulation nor phosphatidylinositol breakdown were affected in the presence of 10 nM ANF. The results further support the role of ANF in the regulation of NE metabolism in the rat hypothalamus. ANF is likely to act as a negative putative neuromodulator inhibiting noradrenergic neurotransmission by signaling through the activation of guanylate cyclase. Thus, ANF may be involved in the regulation of several central as well as peripheral physiological processes such as cardiovascular function, electrolyte and fluid homeostasis, endocrine and neuroendocrine synthesis and secretion, behavior, thirst, appetite and anxiety that are mediated by central noradrenergic activity.
Asunto(s)
Factor Natriurético Atrial/farmacología , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Norepinefrina/biosíntesis , Animales , AMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Activación Enzimática/efectos de los fármacos , Guanilato Ciclasa/metabolismo , Masculino , Norepinefrina/metabolismo , Fosfatidilinositoles/metabolismo , Cloruro de Potasio/farmacología , Ratas , Ratas Wistar , Transducción de Señal , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
The inhibitory effects of bulbocapnine, an aporphine isoquinoline alkaloid, on bovine adrenal tyrosine hydroxylase (TH) were investigated. Bulbocapnine had an inhibitory effect on the enzyme (43.6% inhibition at concentration of 200 microM). Bulbocapnine exhibited uncompetitive inhibition on bovine adrenal TH with a substrate L-tyrosine. The Ki value was found to be 0.20 mM.
Asunto(s)
Aporfinas/farmacología , Inhibidores Enzimáticos/farmacología , Tirosina 3-Monooxigenasa/antagonistas & inhibidores , Glándulas Suprarrenales/efectos de los fármacos , Glándulas Suprarrenales/enzimología , Animales , Bovinos , Cinética , Células PC12 , RatasRESUMEN
This study was designed to determine the behavioral effects of a reduction in catecholamine and indoleamine function in healthy subjects. Eight healthy subjects received the tyrosine hydroxylase inhibitor, alpha-methyl-para-tyrosine (AMPT) in combination with a full-strength tryptophan-depleting amino acid drink during one 4-day test session, and AMPT and tryptophan-supplemented amino acid drink (n = 2), or a 25% strength tryptophan-depleting amino acid drink (n = 6) during a second 4-day test session. The combined administration of AMPT and the tryptophan-free amino acid drink did not produce statistically significant or even clinically noticeable changes in mood among the healthy subjects. The implications of these observations for the monoamine hypotheses of depression are discussed.