RESUMEN
Blood pressure is controlled by endocrine, autonomic, and behavioral responses that maintain blood volume and perfusion pressure at levels optimal for survival. Although it is clear that central angiotensin type 1a receptors (AT1aR; encoded by the Agtr1a gene) influence these processes, the neuronal circuits mediating these effects are incompletely understood. The present studies characterize the structure and function of AT1aR neurons in the lamina terminalis (containing the median preoptic nucleus and organum vasculosum of the lamina terminalis), thereby evaluating their roles in blood pressure control. Using male Agtr1a-Cre mice, neuroanatomical studies reveal that AT1aR neurons in the area are largely glutamatergic and send projections to the paraventricular nucleus of the hypothalamus (PVN) that appear to synapse onto vasopressin-synthesizing neurons. To evaluate the functionality of these lamina terminalis AT1aR neurons, we virally delivered light-sensitive opsins and then optogenetically excited or inhibited the neurons while evaluating cardiovascular parameters or fluid intake. Optogenetic excitation robustly elevated blood pressure, water intake, and sodium intake, while optogenetic inhibition produced the opposite effects. Intriguingly, optogenetic excitation of these AT1aR neurons of the lamina terminalis also resulted in Fos induction in vasopressin neurons within the PVN and supraoptic nucleus. Further, within the PVN, selective optogenetic stimulation of afferents that arise from these lamina terminalis AT1aR neurons induced glutamate release onto magnocellular neurons and was sufficient to increase blood pressure. These cardiovascular effects were attenuated by systemic pretreatment with a vasopressin-1a-receptor antagonist. Collectively, these data indicate that excitation of lamina terminalis AT1aR neurons induces neuroendocrine and behavioral responses that increase blood pressure.SIGNIFICANCE STATEMENT Hypertension is a widespread health problem and risk factor for cardiovascular disease. Although treatments exist, a substantial percentage of patients suffer from "drug-resistant" hypertension, a condition associated with increased activation of brain angiotensin receptors, enhanced sympathetic nervous system activity, and elevated vasopressin levels. The present study highlights a role for angiotensin Type 1a receptor expressing neurons located within the lamina terminalis in regulating endocrine and behavioral responses that are involved in maintaining cardiovascular homeostasis. More specifically, data presented here reveal functional excitatory connections between angiotensin-sensitive neurons in the lamina terminals and vasopressin neurons in the paraventricular nucleus of the hypothalamus, and further indicate that activation of this circuit raises blood pressure. These neurons may be a promising target for antihypertensive therapeutics.
Asunto(s)
Angiotensinas/farmacología , Arginina Vasopresina/metabolismo , Presión Sanguínea/efectos de los fármacos , Hipotálamo/efectos de los fármacos , Vías Nerviosas/efectos de los fármacos , Núcleo Hipotalámico Paraventricular/efectos de los fármacos , Vasoconstrictores/farmacología , Animales , Núcleo Basal de Meynert/efectos de los fármacos , Núcleo Basal de Meynert/metabolismo , Ingestión de Líquidos/efectos de los fármacos , Genes fos/efectos de los fármacos , Ácido Glutámico/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Optogenética , Receptor de Angiotensina Tipo 1/efectos de los fármacos , Receptores de Vasopresinas/efectos de los fármacos , Sodio en la DietaRESUMEN
Curcumae radix is the dry root of Curcuma longa L. (turmeric) that can be used either as a spice or traditional medicine. The aim of this study was to investigate the survival benefits and the anti-metastatic activity of curcumae radix extract (CRE) in MCF7 cells and in MMTV-PyMT transgenic mice-a mouse model of breast cancer metastasis. In vitro wound scratch assay revealed that CRE treatment inhibited cell motility and cell migration in a dose-dependent manner. To investigate the effect of CRE in breast cancer metastasis, MMTV-PyMT transgenic female virgin mice were used and randomly divided into two groups. For survival curve analysis, CRE was administered in a dose of 50 mg/kg to 8â»20-week-old mice. Interestingly, CRE treatment significantly increased the median and prolonged survival of MMTV-PyMT mice. Furthermore, CRE treatment decreased tumor burden and inhibited cell proliferation in primary breast tumor, and also suppressed mammary tumor-derived lung metastasis. The size of the lung metastases substantially decreased in the CRE-treated group compared with the ones in the control group. Curcumae radix extract showed anti-metastatic activity through regulating the expression of metastasis markers including C-C Chemokine Receptor Type 7, Matrix Metalloproteinase 9 and the proto-oncogenes c-fos and c-jun. We demonstrated that these metastatic regulators were decreased when CCR7 expression was suppressed in MCF7 cells transfected with CCR7 siRNA. The results of this study show that curcumae radix exerts antitumor and anti-metastatic activities, and we suggest that curcumae radix might be a potential supplement for the treatment and prevention of breast cancer metastasis.
Asunto(s)
Antineoplásicos/farmacología , Curcuma , Neoplasias Pulmonares/prevención & control , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Metástasis de la Neoplasia/prevención & control , Extractos Vegetales/farmacología , Receptores CCR7/efectos de los fármacos , Animales , Femenino , Genes fos/efectos de los fármacos , Genes jun/efectos de los fármacos , Neoplasias Pulmonares/secundario , Neoplasias Mamarias Experimentales/patología , Metaloproteinasa 9 de la Matriz/efectos de los fármacos , Ratones , Ratones Transgénicos , Raíces de Plantas , Receptores CCR7/biosíntesisRESUMEN
BACKGROUND The aim of this study was to investigate the effects of electroacupuncture (EA) on expression of the D1 receptor (D1R), phosphorylation of extracellular-regulated protein kinase 1/2 (p-ERK1/2) and c-Fos in the insular cortex (IC) of ketamine-addicted rats. MATERIAL AND METHODS Sprague-Dawley rats were randomly divided into 7 groups: the normal group, the normal saline (NS) group, the ketamine (Ket) group, the U0126+Ket group, the SCH23390+Ket group, the Ket+acupoints EA (EA1) group, and the Ket+ non-acupoints EA (EA2) group. We used immunohistochemistry to detect the expression of D1R, p-ERK1/2, and c-Fos. We also used Nissl staining techniques to study the morphology of IC neurons. RESULTS Our study demonstrated that the ketamine group had sparsely distributed neurons, large intracellular vacuoles, nuclei shift, and unclear nucleolus. The number of Nissl-positive (neuronal) cells in the ketamine group were decreased than in the normal group. Our results also indicated that there was significantly lower expression of D1R, p-ERK1/2, and c-Fos in the IC of the U0126+Ket group, SCH23390+Ket group, and Ket+EA1 group as compared with that of the Ket group. CONCLUSIONS Ketamine addiction induces c-Fos overexpression in the IC by increasing the expression of D1R and p-ERK1/2. Acupoints EA downregulate D1R and p-ERK1/2 by reducing the overexpression of c-Fos.
Asunto(s)
Corteza Cerebral/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Receptores de Dopamina D1/efectos de los fármacos , Puntos de Acupuntura , Animales , Butadienos/farmacología , Electroacupuntura/métodos , Genes fos/efectos de los fármacos , Genes fos/fisiología , Ketamina/farmacología , Sistema de Señalización de MAP Quinasas/fisiología , Masculino , Neuronas/efectos de los fármacos , Nitrilos/farmacología , Fosforilación , Ratas , Ratas Sprague-Dawley , Receptores de Dopamina D1/metabolismoRESUMEN
Abstract Introduction: Salicylate at high doses induces tinnitus in humans and experimental animals. However, the mechanisms and loci of action of salicylate in inducing tinnitus are still not well known. The expression of Immediate Early Genes (IEG) is traditionally associated with long-term neuronal modifications but it is still not clear how and where IEGs are activated in animal models of tinnitus. Objectives: Here we investigated the expression of c-fos and Egr-1, two IEGs, in the Dorsal Cochlear Nucleus (DCN), the Inferior Colliculus (IC), and the Posterior Ventral Cochlear Nucleus (pVCN) of rats. Methods: Rats were treated with doses known to induce tinnitus in rats (300 mg/kg i.p. daily, for 3 days), and c-fos and Egr-1 protein expressions were analyzed using western blot and immunocytochemistry. Results: After administration of salicylate, c-fos protein expression increased significantly in the DCN, pVCN and IC when assayed by western blot. Immunohistochemistry staining showed a more intense labeling of c-fos in the DCN, pVCN and IC and a significant increase in c-fos positive nuclei in the pVCN and IC. We did not detect increased Egr-1 expression in any of these areas. Conclusion: Our data show that a high dose of salicylate activates neurons in the DCN, pVCN and IC. The expression of these genes by high doses of salicylate strongly suggests that plastic changes in these areas are involved in the genesis of tinnitus.
Resumo Introdução: Salicilato em doses elevadas induz zumbido nos seres humanos e em animais experimentais. No entanto, os mecanismos e loci de ação do salicilato na indução de zumbido ainda não são bem conhecidos. A expressão dos genes precoces imediatos (GPIs) está tradicionalmente associada a alterações neuronais em longo prazo, mas ainda não está claro como e onde os GPIs são ativados em modelos animais de zumbido. Objetivos: No presente estudo investigamos a expressão de c-fos e Egr-1, dois GPIs, no núcleo coclear dorsal (NCD), colículo inferior (CI) e núcleo coclear ventral posterior (NCVp) de ratos. Métodos: Os ratos foram tratados com doses que, conhecidamente, induzem zumbido em ratos (300 mg/kg IP/dia, por três dias) e as expressões das proteínas c-fos e Egr-1 foram analisadas por meio de Western blot e imunoistoquímica. Resultados: Após a administração de salicilato, a expressão da proteína c-fos aumentou significativamente no NCD, NCVp e CI, quando analisados por Western blot. A coloração imunoistoquímica mostrou uma marcação mais intensa de c-fos no NCD, NCVp e CI e um aumento significativo de núcleos positivos de c-fos no NCVp e CI. Não detectamos aumento da expressão de Egr-1 em qualquer dessas áreas. Conclusão: Nossos dados mostram que uma dose alta de salicilato ativa neurônios no NCD, NCVp e CI. A expressão desses genes por doses altas de salicilato sugere que as alterações plásticas nessas áreas estão envolvidas na gênese do zumbido.
Asunto(s)
Animales , Masculino , Ratas , Colículos Inferiores/efectos de los fármacos , Salicilatos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Genes Inmediatos-Precoces/efectos de los fármacos , Núcleo Coclear/efectos de los fármacos , Salicilatos/administración & dosificación , Western Blotting , Genes fos/efectos de los fármacos , Ratas Wistar , Relación Dosis-Respuesta a Droga , Proteína 1 de la Respuesta de Crecimiento Precoz/efectos de los fármacosRESUMEN
INTRODUCTION: Salicylate at high doses induces tinnitus in humans and experimental animals. However, the mechanisms and loci of action of salicylate in inducing tinnitus are still not well known. The expression of Immediate Early Genes (IEG) is traditionally associated with long-term neuronal modifications but it is still not clear how and where IEGs are activated in animal models of tinnitus. OBJECTIVES: Here we investigated the expression of c-fos and Egr-1, two IEGs, in the Dorsal Cochlear Nucleus (DCN), the Inferior Colliculus (IC), and the Posterior Ventral Cochlear Nucleus (pVCN) of rats. METHODS: Rats were treated with doses known to induce tinnitus in rats (300mg/kg i.p. daily, for 3 days), and c-fos and Egr-1 protein expressions were analyzed using western blot and immunocytochemistry. RESULTS: After administration of salicylate, c-fos protein expression increased significantly in the DCN, pVCN and IC when assayed by western blot. Immunohistochemistry staining showed a more intense labeling of c-fos in the DCN, pVCN and IC and a significant increase in c-fos positive nuclei in the pVCN and IC. We did not detect increased Egr-1 expression in any of these areas. CONCLUSION: Our data show that a high dose of salicylate activates neurons in the DCN, pVCN and IC. The expression of these genes by high doses of salicylate strongly suggests that plastic changes in these areas are involved in the genesis of tinnitus.
Asunto(s)
Núcleo Coclear/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Genes Inmediatos-Precoces/efectos de los fármacos , Colículos Inferiores/efectos de los fármacos , Salicilatos/farmacología , Animales , Western Blotting , Relación Dosis-Respuesta a Droga , Proteína 1 de la Respuesta de Crecimiento Precoz/efectos de los fármacos , Genes fos/efectos de los fármacos , Masculino , Ratas , Ratas Wistar , Salicilatos/administración & dosificaciónRESUMEN
RATIONALE: Ethnopharmacology has documented hundreds of psychoactive plants awaiting exploitation for drug discovery. A robust and inexpensive in vivo system allowing systematic screening would be critical to exploiting this knowledge. OBJECTIVE: The objective of this study was to establish a cheap and accurate screening method which can be used for testing psychoactive efficacy of complex mixtures of unknown composition, like plant crude extracts. METHODS: We used automated recording of zebrafish larval swimming behavior during light vs. dark periods which we reproducibly altered with an anxiogenic compound, pentylenetetrazole (PTZ). First, we reversed this PTZ-altered swimming by co-treatment with a well-defined synthetic anxiolytic drug, valproic acid (VPA). Next, we aimed at reversing it by adding crude root extracts of Valeriana officinalis (Val) from which VPA was originally derived. Finally, we assessed how expression of neural activity-regulated genes (c-fos, npas4a, and bdnf) known to be upregulated by PTZ treatment was affected in the presence of Val. RESULTS: Both VPA and Val significantly reversed the PTZ-altered swimming behaviors. Noticeably, Val at higher doses was affecting swimming independently of the presence of PTZ. A strong regulation of all three neural-activity genes was observed in Val-treated larvae which fully supported the behavioral results. CONCLUSIONS: We demonstrated in a combined behavioral-molecular approach the strong psychoactivity of a natural extract of unknown composition made from V. officinalis. Our results highlight the efficacy and sensitivity of such an approach, therefore offering a novel in vivo screening system amenable to high-throughput testing of promising ethnobotanical candidates.
Asunto(s)
Antimaníacos/farmacología , Conducta Animal/efectos de los fármacos , Encéfalo/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Pentilenotetrazol/farmacología , Extractos Vegetales/farmacología , Natación/fisiología , Valeriana , Ácido Valproico/farmacología , Animales , Ansiolíticos/farmacología , Biomarcadores/metabolismo , Encéfalo/metabolismo , Quimioterapia Combinada , Genes fos/efectos de los fármacos , Hipnóticos y Sedantes/farmacología , Larva , Proteínas Proto-Oncogénicas c-fos/metabolismo , Pez Cebra/fisiologíaRESUMEN
BACKGROUND: The capsaicin and heat responsive ion channel TRPV1 is expressed on trigeminal nociceptive neurons and has been implicated in the pathophysiology of migraine attacks. Here we investigate the efficacy of two TRPV1 channel antagonists in blocking trigeminal activation using two in vivo models of migraine. METHODS: Male Sprague-Dawley rats were used to study the effects of the TRPV1 antagonists JNJ-38893777 and JNJ-17203212 on trigeminal activation. Expression of the immediate early gene c-fos was measured following intracisternal application of inflammatory soup. In a second model, CGRP release into the external jugular vein was determined following injection of capsaicin into the carotid artery. RESULTS: Inflammatory up-regulation of c-fos in the trigeminal brain stem complex was dose-dependently and significantly reduced by both TRPV1 antagonists. Capsaicin-induced CGRP release was attenuated by JNJ-38893777 only in higher dosage. JNJ-17203212 was effective in all doses and fully abolished CGRP release in a time and dose-dependent manner. CONCLUSION: Our results describe two TRPV1 antagonists that are effective in two in vivo models of migraine. These results suggest that TRPV1 may play a role in the pathophysiological mechanisms, which are relevant to migraine.
Asunto(s)
Aminopiridinas/uso terapéutico , Modelos Animales de Enfermedad , Trastornos Migrañosos/tratamiento farmacológico , Piperazinas/uso terapéutico , Canales Catiónicos TRPV/antagonistas & inhibidores , Aminopiridinas/farmacología , Animales , Capsaicina/toxicidad , Relación Dosis-Respuesta a Droga , Genes fos/efectos de los fármacos , Masculino , Trastornos Migrañosos/inducido químicamente , Trastornos Migrañosos/metabolismo , Piperazinas/farmacología , Ratas , Ratas Sprague-Dawley , Resultado del Tratamiento , Regulación hacia Arriba/efectos de los fármacosRESUMEN
The classical benzodiazepine diazepam (DZ) induces anxiolysis at low doses and sedation and hypnosis at higher doses. Different brain areas and neuronal populations most likely mediate these different behavioral effects. We used c-Fos immunohistochemistry as an indirect way to study neuronal activation or inhibition induced by DZ at anxiolytic and sedative doses (0.5 and 5mg/kg, respectively) in various brain areas involved in anxiety, arousal, sedation and addiction in C57BL/6J mice. We also focused on the two neuronal populations, orexinergic and dopaminergic neuronal populations, with the help of double-immunohistochemistry using c-Fos and orexin-A antibodies and c-Fos and tyrosine hydroxylase antibodies. We found that different brain areas of unhabituated mice reacted differently to the mild stress induced by vehicle injection. Also the response to anxiolytic or sedative doses of DZ differed between the areas, suggesting that distinct brain areas mediate the behavioral effects of low and high DZ doses. Our findings propose a role for inhibition of orexin neurons in the anxiolytic and sleep-promoting effects of DZ. In addition, the activation of central amygdala neurons by DZ treatment was associated with anxiolytic and sedative effects. On the other hand, the ventral hippocampus, basolateral amygdala, ventral tegmental area and prefrontal cortex were sensitive even to the mild injection stress, but not to the anxiolytic dose of DZ.
Asunto(s)
Ansiolíticos/farmacología , Diazepam/farmacología , Genes fos/efectos de los fármacos , Hipnóticos y Sedantes/farmacología , Péptidos y Proteínas de Señalización Intracelular/fisiología , Neuronas/fisiología , Neuropéptidos/fisiología , Envejecimiento/fisiología , Amígdala del Cerebelo/citología , Amígdala del Cerebelo/efectos de los fármacos , Amígdala del Cerebelo/metabolismo , Animales , Ansiedad/psicología , Neuronas Dopaminérgicas/fisiología , Expresión Génica/efectos de los fármacos , Hipocampo/citología , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Hipotálamo/fisiología , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Neuronas/efectos de los fármacos , Neuropéptidos/metabolismo , Orexinas , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Nocistatin and nociceptin/orphanin FQ (N/OFQ) are two neuropeptides which may have opposite effects in several biological functions but their neuro-anatomical sites of interaction are not fully clear. We investigated interaction between the effect of intracerebroventricular (i.c.v.) injection of nocistatin and N/OFQ, on c-Fos expression in the mouse thalamus, using c-Fos immunohistochemistry. We found that co-injection of nocistatin with N/OFQ significantly modulates c-Fos expression in the thalamus. The present study strongly suggests that "Nocistatin-Nociceptin" interaction system in the thalamus may be the promising neuromodulatory sites in the investigation of unlocking their possible therapeutic circuit in nociception, memory and anxiety.
Asunto(s)
Regulación de la Expresión Génica , Genes fos/fisiología , Péptidos Opioides/administración & dosificación , Tálamo/metabolismo , Animales , Genes fos/efectos de los fármacos , Humanos , Inyecciones Intraventriculares , Ratones , Péptidos Opioides/fisiología , Tálamo/efectos de los fármacos , NociceptinaRESUMEN
It has been reported that the sedative component of pentobarbital is mediated by GABA receptors in an endogenous sleep pathway and the ventrolateral preoptic area (VLPO)-tuberomammillary nucleus (TMN) or VLPO-dorsal raphe nucleus (DRN) neural circuit is important in the sedative response to pentobarbital. Our previous findings indicated that the VLPO-TMN neuronal circuit may play crucial part in the augmentative effect of diltiazem on pentobarbital sleep and the serotonergic system may be involved. This study was designed to investigate the role of DRN and the serotonergic receptors 5-HT(1A) and 5-HT(2A/2C) in the augmentative effect of diltiazem on pentobarbital-induced hypnosis in rats. The results showed that diltiazem (5mg/kg, i.g.) significantly reversed pentobarbital-induced (35 mg/kg, i.p.) reduction of c-Fos expression in 5-HT neurons of DRNV (at -7.5mm Bregma), DRND, DRNVL and MRN (at -8.0mm Bregma). However it did not influence this reducing effect of pentobarbital on non-5-HT neurons either in DRN or in MRN. Moreover, the effect of diltiazem (1 or 2mg/kg, i.g.) on pentobarbital-induced (35 mg/kg, i.p.) hypnosis was significantly inhibited by 5-HT(1A) agonist 8-OH-DPAT (0.5mg/kg, i.p.) and 5-HT(2A/2C) agonist DOI (0.5mg/kg, i.p.), and potentiated by 5-HT(1A) antagonist p-MPPI (2mg/kg, i.p.) and 5-HT(2A/2C) antagonist ritanserin (2mg/kg, i.p.), respectively. From these results, it should be presumed that the augmentative effect of diltiazem on pentobarbital-induced sleep may be related to 5-HT(1A) and 5-HT(2A/2C) receptors, and DRN may be involved. In addition, it also suggested that the DRN may play a multi-modulating role in sleep-wake regulation rather than being recognized simply as arousal nuclei.
Asunto(s)
Bloqueadores de los Canales de Calcio/farmacología , Diltiazem/farmacología , Hipnóticos y Sedantes/farmacología , Pentobarbital/farmacología , Núcleos del Rafe/fisiología , Receptor de Serotonina 5-HT1A/efectos de los fármacos , Receptor de Serotonina 5-HT2A/efectos de los fármacos , Receptor de Serotonina 5-HT2C/efectos de los fármacos , Animales , Encéfalo/citología , Encéfalo/efectos de los fármacos , Recuento de Células , Sinergismo Farmacológico , Electroencefalografía/efectos de los fármacos , Electromiografía/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Genes fos/efectos de los fármacos , Inmunohistoquímica , Masculino , Polisomnografía , Núcleos del Rafe/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Neuronas Serotoninérgicas/efectos de los fármacos , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacologíaRESUMEN
The pathogenesis of necrotizing enterocolitis (NEC) is unknown. Ischemia and reperfusion (I/R) injury have been considered to be major contributing factors. More recent reports have noted that apoptosis is a significant and perhaps the principal contributor to cell death after I/R injury. Recent studies have revealed that activator protein 1 (AP-1) family proteins including c-Fos and c-Jun potentially induce either the proliferation or apoptosis of the cells in the brain, heart, kidney, and liver. c-Fos and c-Jun expression has also been reported to be upregulated in postischemic intestinal epithelial cells (IECs). Heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) is a potent cytoprotective factor in various pathologic conditions and plays a pivotal role in mediating the earliest cellular responses to injury. This study aims to examine whether HB-EGF, a proven intestinal cytoprotective molecule, exerts its protective effects through modulation of AP-1 transcription factor after intestinal I/R injury. Thirty rats were randomly divided into the following 5 groups: (1) normal control group; (2) ischemia group; (3) I/R group; (4) ischemia group with HB-EGF (400 µg/kg), and (5) I/R group with HG-EGF (400 µg/kg). c-Fos and c-Jun messenger RNAs and protein levels were determined by real-time quantitative polymerase chain reaction (PCR) and Western analyses, respectively. Statistical analysis was performed using ANOVA with Dunn's test. The messenger RNA levels of the c-Fos and c-Jun increased after intestinal ischemia or the intestinal reperfusion phase. HB-EGF pretreatment significantly decreased c-Fos and c-Jun messenger RNAs. The expression of protein levels of c-Fos and c-Jun were correlation with the expression of messenger RNA level. HB-EGF intestinal cytoprotection is mediated, in part, by downregulation of the expression of AP-1 transcription factor after intestinal I/R injury.
Asunto(s)
Péptidos y Proteínas de Señalización Intercelular/farmacología , Mucosa Intestinal/metabolismo , Intestinos/efectos de los fármacos , Daño por Reperfusión , Factor de Transcripción AP-1/genética , Animales , Citoprotección/efectos de los fármacos , Citoprotección/genética , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Evaluación Preclínica de Medicamentos , Regulación de la Expresión Génica/efectos de los fármacos , Genes fos/efectos de los fármacos , Genes jun/efectos de los fármacos , Factor de Crecimiento Similar a EGF de Unión a Heparina , Intestinos/irrigación sanguínea , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/genética , Daño por Reperfusión/metabolismo , Daño por Reperfusión/fisiopatología , Factor de Transcripción AP-1/metabolismoRESUMEN
Estrogen may be involved in psychosis by an interaction with central dopaminergic activity. Aromatase knockout mice are unable to produce estrogen and have been shown to display altered behavioural responses and effects of the dopamine releaser, amphetamine. This study investigates the effect of gonadal status on amphetamine-induced c-fos expression in the brains of female aromatase knockout and wildtype mice. Six groups of mice were treated intraperitoneally with saline or 5mg/kg amphetamine. Fos immunoreactivity was assessed in the cingulate cortex, caudate putamen and nucleus accumbens. Aromatase knockout mice showed markedly reduced amphetamine-induced Fos immunoreactivity compared to wildtype mice. However, the amphetamine response was restored in aromatase-knockout mice after ovariectomy, which reduced this effect in wildtype controls. Estrogen supplementation reversed the effect of ovariectomy in wildtype mice but had no additional significant effect in aromatase-knockout mice. These results indicate that mechanisms involved in amphetamine-induced c-fos expression are altered in aromatase knockout mice and that the primary hormone involved in this effect is not estrogen, but may be another factor released from the ovaries, such as an androgen. These results provide new insight into the effect of gonadal hormones on amphetamine induced c-fos expression in this mouse model of estrogen deficiency. These results could be important for our understanding of the role of sex steroid hormones in psychosis.
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Anfetamina/farmacología , Aromatasa/genética , Expresión Génica/efectos de los fármacos , Genes fos/efectos de los fármacos , Animales , Peso Corporal/efectos de los fármacos , Femenino , Genes fos/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Modelos Biológicos , OvariectomíaRESUMEN
AIM OF THE STUDY: Asian scorpion Buthus martensi Karsch (BmK) is widely used to treat neurological symptoms, especially chronic pain, in traditional Chinese medicine for thousands of years. BmK AS, a polypeptide from BmK venom, could produce peripheral potent anti-nociceptive effects in rats. In the present study, spinal anti-nociceptive effects of BmK AS were investigated in rat formalin test. MATERIALS AND METHODS: Spinal anti-nociceptive activity of BmK AS was studied using formalin test in rats. BmK AS in doses of 0.02, 0.1 and 0.5 microg was administered intrathecally before formalin injection 10 min. The suppression by intrathecal injection of BmK AS on formalin-induced spontaneous nociceptive behaviors and spinal c-Fos expression were investigated. RESULTS: Intrathecal injection of BmK AS markedly reduced formalin-evoked biphasic spontaneous nociceptive behaviors in a dose-dependent manner. Formalin-induced c-Fos expression could be dose-dependently inhibited by BmK AS in superficial (I-II), the nucleus proprius (III and IV) and deep (V-VI) dorsal horn laminae, but not in the ventral gray laminae (VII-X) of lumbar spinal cord. The suppression by BmK AS on c-Fos expression in superficial laminaes was much stronger than that in deep laminaes. CONCLUSION: The present study demonstrates that BmK AS is capable of producing remarkable anti-nociceptive effects not only in periphery but also in spinal cord.
Asunto(s)
Analgésicos/farmacología , Dimensión del Dolor/efectos de los fármacos , Péptidos/farmacología , Venenos de Escorpión/química , Animales , Relación Dosis-Respuesta a Droga , Formaldehído , Expresión Génica/efectos de los fármacos , Genes fos/efectos de los fármacos , Inmunohistoquímica , Inyecciones Espinales , Masculino , Péptidos/administración & dosificación , Péptidos/química , Ratas , Ratas Sprague-Dawley , Venenos de Escorpión/administración & dosificación , Venenos de Escorpión/farmacologíaRESUMEN
We have developed a model to study central changes following inflammation of the tooth pulp in the ferret and have examined Fos expression in the trigeminal nucleus following stimulation of non-inflamed and inflamed tooth pulps. The aim of this study was to establish the ability of this model to predict analgesic efficacy in clinical studies of inflammatory pain. We addressed this by assessing the effects of the neurokinin-1 receptor antagonist GR205171A and ibuprofen on Fos expression following stimulation of the inflamed pulp and comparing this with known analgesic efficacy. Adult ferrets were prepared under anaesthesia to allow tooth pulp stimulation, recording from the digastric muscle and intravenous injections at a subsequent experiment. In some animals pulpal inflammation was induced, by introducing human caries into a deep buccal cavity. After 5 days, animals were reanaesthetised, treated with vehicle, GR205171A or ibuprofen and the teeth were stimulated at ten times the threshold of the jaw-opening reflex. Stimulation of all tooth pulps induced ipsilateral Fos in trigeminal subnuclei caudalis and oralis. GR205171A had no significant effect on Fos expression in the trigeminal nucleus of animals with either non-inflamed or inflamed tooth pulps. Ibuprofen reduced Fos expression in the trigeminal nucleus and this effect was most marked in animals with pulpal inflammation. These results differ from those previously described using a range of other animal models, but agree with known clinical efficacy of neurokinin-1 receptor antagonists and ibuprofen. Therefore this model is likely to be of use in accurately predicting the analgesic efficacy of novel compounds.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Pulpa Dental/inervación , Regulación de la Expresión Génica/efectos de los fármacos , Genes fos/efectos de los fármacos , Ibuprofeno/farmacología , Proteínas del Tejido Nervioso/biosíntesis , Antagonistas del Receptor de Neuroquinina-1 , Fragmentos de Péptidos/farmacología , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Pulpitis/fisiopatología , Sustancia P/análogos & derivados , Núcleos del Trigémino/metabolismo , Animales , Antiinflamatorios no Esteroideos/uso terapéutico , Diente Canino , Inhibidores de la Ciclooxigenasa/farmacología , Inhibidores de la Ciclooxigenasa/uso terapéutico , Evaluación Preclínica de Medicamentos , Estimulación Eléctrica , Hurones , Ibuprofeno/uso terapéutico , Proteínas del Tejido Nervioso/genética , Fragmentos de Péptidos/uso terapéutico , Pulpitis/tratamiento farmacológico , Pulpitis/genética , Receptores de Neuroquinina-1/fisiología , Sustancia P/farmacología , Sustancia P/uso terapéutico , Neuralgia del Trigémino/tratamiento farmacológico , Neuralgia del Trigémino/etiología , Neuralgia del Trigémino/fisiopatología , Núcleos del Trigémino/fisiopatologíaRESUMEN
OBJECTIVE: To study the effects of selenium on DNA damage, apoptosis and c-myc, c-fos, and c-jun expression in rat hepatocytes. METHODS: Sodium selenite at the doses of 5, 10, and 20 micromol/kg was given to rats by i.p. and there were 5 male SD rats in each group. Hepatocellular DNA damage was detected by single cell gel electrophoresis (or comet assay). Hepatocellular apoptosis was determined by TUNEL (TdT-mediated dUTP nick end labelling) and flow cytometry. C-myc, c-fos, and c-jun expression in rat hepatocytes were assayed by Northern dot hybridization. C-myc, c-fos, and c-jun protein were detected by immunohistochemical method. RESULTS: At the doses of 5, 10, and 20 micromol/kg, DNA damage was induced by sodium selenite in rat hepatocytes and the rates of comet cells were 34.40%, 74.80%, and 91.40% respectively. Results also showed an obvious dose-response relationship between the rates of comet cells and the doses of sodium selenite (r=0.9501, P<0.01). Sodium selenite at the doses of 5, 10, and 20 micromol/kg caused c-myc, c-fos, and c-jun overexpression obviously. The positive brown-yellow signal for proteins of c-myc, c-fos, and c-jun was mainly located in the cytoplasm of hepatocytes with immunohistochemical method. TUNEL-positive cells were detected in selenium-treated rat livers. Apoptotic rates (%) of selenium-treated liver cells at the doses of 5, 10, and 20 micromol/kg were (3.72 +/- 1.76), (5.82 +/- 1.42), and (11.76 +/- 1.87) respectively, being much higher than those in the control. Besides an obvious dose-response relationship between apoptotic rates and the doses of sodium selenite (r=0.9897, P<0.01), these results displayed a close relationship between DNA damage rates and apoptotic rates, and the relative coefficient was 0.9021, P<0.01. CONCLUSION: Selenium at 5-20 micromol/kg can induce DNA damage, apoptosis, and overexpression of c-myc, c-fos, and c-jun in rat hepatocytes.
Asunto(s)
Apoptosis/efectos de los fármacos , Daño del ADN , Genes fos/efectos de los fármacos , Genes jun/efectos de los fármacos , Genes myc/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Selenio/farmacología , Animales , Northern Blotting , Ensayo Cometa , Relación Dosis-Respuesta a Droga , Genes fos/genética , Genes jun/genética , Genes myc/genética , Hepatocitos/patología , Masculino , Hibridación de Ácido Nucleico , Ratas , Ratas Sprague-Dawley , Selenito de Sodio/farmacologíaRESUMEN
The anti-inflammatory effect of the medicinal plant, Commiphora mukul gum was studied in peripheral blood mononuclear cells (PBMC). Bioassay-guided fractionation using conventional solvent extraction procedures, subsequent column fractionation, followed by monitoring specific activity in PBMC led to the isolation of a lead compound. Both crude ethyl acetate extract and the lead compound, thus isolated, showed inhibitory effect on proliferative response of PBMC in mitogenic lymphocyte proliferation and MLR assays. Further studies on inflammatory mediators such as IFN-gamma, IL-12, TNF-alpha, IL-1beta and NO showed down regulation, whereas no inhibition was observed in the case of anti-inflammatory cytokine IL-10. Immunoblot analysis revealed the inhibitory effect of crude ethyl acetate extract on phosphorylation of all the three mitogen activated protein kinases (MAPK) such as ERK, JNK and p38 MAPK. In contrast treatment with pure compound showed no inhibitory effect on ERK. c-fos and c-jun mRNA levels were also reduced in PMA stimulated cells on treatment with crude extract and pure compound. This reduction in c-fos and c-jun levels, when taken together with inhibition of MAPK activation, provides a possible mechanism by which both crude ethyl acetate extract and purified compound isolated from C. mukul exert its action.
Asunto(s)
Commiphora/química , Interleucina-1/biosíntesis , Interleucina-2/biosíntesis , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/biosíntesis , Acetatos , Animales , Western Blotting , Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Genes fos/efectos de los fármacos , Genes jun/efectos de los fármacos , Humanos , Indicadores y Reactivos , Interleucina-1/antagonistas & inhibidores , Interleucina-2/antagonistas & inhibidores , Prueba de Cultivo Mixto de Linfocitos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Fitohemaglutininas/farmacología , Extractos Vegetales/farmacología , ARN Mensajero/biosíntesis , Solventes , Sales de Tetrazolio , Tiazoles , Factor de Necrosis Tumoral alfa/antagonistas & inhibidoresRESUMEN
The renin-angiotensin system plays an important role in cardiovascular control. Intracerebroventricular (i.c.v.) angiotensin (ANG) II causes a reliable pressor response in the fetus at 90% gestation. To determine the roles of brain AT1 and AT2 receptors in this response, the effects of the central AT1 and AT2 receptor antagonists losartan and PD123319 were investigated in chronically prepared near-term ovine fetuses. Losartan at 0.5 mg/kg (i.c.v.) abolished central ANG II-induced pressor responses. High-dose losartan (5 mg/kg, i.c.v.) showed a potentiation of the pressor response to i.c.v. ANG II, accompanied by bradycardia. Associated with the pressor responses, c-fos expression in the cardiovascular controlling areas was significantly different between the low and high doses of losartan. These areas included the subfornical organ, median preoptic nucleus, organum vasculosum of the lamina terminalis, and paraventricular nuclei in the forebrain, and the tractus solitarius nuclei, lateral parabrachial nuclei in the hindbrain. Low-dose losartan markedly reduced c-fos in these areas after i.c.v. ANG II, while the high-dose losartan together with ANG II elicited a much stronger FOS-immunoreactivity in these areas than that induced by i.c.v. ANG II alone. This is a novel finding, that c-fos expression in the brain can be both activated and inhibited under the same condition. Central ANG II-induced fetal pressor responses were not altered by PD123319 (0.8 mg/kg). These results indicate that i.c.v. losartan at a high and a low dose has strikingly different effects on central ANG II-induced pressor responses in fetuses at late gestation, and that the AT1 mechanism plays an important role in fetal cardiovascular regulation.
Asunto(s)
Bloqueadores del Receptor Tipo 1 de Angiotensina II/administración & dosificación , Angiotensina II/efectos de los fármacos , Genes fos/efectos de los fármacos , Losartán/administración & dosificación , Prosencéfalo/efectos de los fármacos , Análisis de Varianza , Angiotensina II/administración & dosificación , Angiotensina II/metabolismo , Animales , Presión Sanguínea/efectos de los fármacos , Sistema Cardiovascular/efectos de los fármacos , Sistema Cardiovascular/embriología , Sistema Cardiovascular/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Feto/efectos de los fármacos , Genes fos/fisiología , Edad Gestacional , Homeostasis/efectos de los fármacos , Homeostasis/fisiología , Inyecciones Intraventriculares , Embarazo , Prosencéfalo/embriología , Prosencéfalo/metabolismo , Ovinos , Estadísticas no ParamétricasRESUMEN
Twenty 5-week-old male Wistar rats were divided into two groups: one group was fed a fish oil-deficient diet and the other group was fed the same diet supplemented with per orally administered docosahexaenoic acid (DHA) for 12 weeks. Six weeks after the start of the administration of DHA, rats were trained for 6 weeks to acquire a reward at the end of each of four arms of an eight-arm radial maze. On completion of the radial maze task, the Fos expression in the hippocampus was examined immunohistochemically. Chronic DHA administration significantly reduced the number of reference and working memory errors. The number of Fos-positive neurons in the CA1 hippocampus significantly increased in DHA-treated rats compared with control rats, demonstrating a statistically significant negative correlation with the number of reference memory errors. These results suggest that the DHA-induced improvement in spatial cognition is associated with increased Fos expression in the CA1 hippocampus.
Asunto(s)
Dieta , Ácidos Docosahexaenoicos/farmacología , Genes fos/efectos de los fármacos , Hipocampo/metabolismo , Memoria/efectos de los fármacos , Percepción Espacial/efectos de los fármacos , Animales , Expresión Génica/efectos de los fármacos , Hipocampo/efectos de los fármacos , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Memoria a Corto Plazo/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Ratas , Ratas WistarRESUMEN
Mice lacking normal vestibular gravity reception show altered homeostatic, circadian and autonomic responses to hypergravity (+G) exposure. Using c-Fos as a marker of neuronal activation, the current study identifies CNS nuclei that may be critical for initiating and integrating such responses to changes in vestibular signaling. This experiment utilized the mutant C57BL/6JEi-het mouse (het), which lacks macular otoconia and thus gravity receptor function. Following 2 h of 2G (2x Earth's gravity) exposure (via centrifugation) the neuronal responses of the het mice were compared with wildtype mice similarly exposed to 2G, as well as het and wildtype 1G controls. Wildtype mice exposed to 2G demonstrated robust c-Fos expression in multiple autonomic, hypothalamic and limbic nuclei, including: the lateral septum, bed nucleus of the stria terminalis, amygdala, paraventricular hypothalamus, dorsomedial hypothalamus, arcuate, suprachiasmatic hypothalamus, intergeniculate leaflet, dorsal raphe, parabrachial and locus coeruleus. The het mice exposed to 2G demonstrated little to null c-Fos expression in these nuclei with a few exceptions and, in general, a similar pattern of c-Fos to 1G controls. Data from this study further support the existence of a complex and extensive influence of the neurovestibular system on homeostatic, circadian and possibly autonomic regulatory systems.
Asunto(s)
Sistema Nervioso Autónomo/fisiología , Hipotálamo/fisiología , Sistema Límbico/fisiología , Mácula Lútea/fisiología , Animales , Plexo Braquial/fisiología , Genes fos/efectos de los fármacos , Hipergravedad , Hipotálamo/anatomía & histología , Inmunohistoquímica , Sistema Límbico/anatomía & histología , Mácula Lútea/anatomía & histología , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes Neurológicos , Vestíbulo del Laberinto/fisiologíaRESUMEN
The Toxic Oil Syndrome (TOS) was a toxic epidemic disease, related to the consumption of rapeseed oil denatured with aniline that affected more than 20,000 people in Spain and resulted in more than 330 deaths after its sudden appearance in 1981. It has been reported that the fatty acid esters of 3-(N-phenylamino)-1,2-propanediol (PAP) have shown a strong association with TOS. These PAP-esters could be absorbed and metabolized in a similar way than phospholipids. This is of interest because some products of phospholipid metabolism are important mediators in downstream pathways involved in the regulation of different nuclear factors. In particular, phospholipase D activity is involved in the activation of c-fos. Thus, we have investigated the effect of different PAP-esters in the induction of c-fos in lung fibroblasts. Results indicate that PAP-esters rapidly induced the expression of c-fos in a dose-dependent manner. In addition, both butanol and propranolol prevent this induction pointing to the involvement of phospholipase D in this activation. These results suggest that deregulation of some nuclear factors such as AP-1 could be involved in the pathogenesis of TOS.