RESUMO
The compatibility of Rhizoma Corydalis(Yuanhu,YH) and Radix Angelicae dahuricae (Baizhi,BZ) as a herb pair Yuanhu-Baizhi(YB) can produce synergistic analgesic effect. However, the underlying mechanism of this herb pair compatibility is not elucidated yet. A LC-MS/MS method combined with in vivo microdialysis sampling from awaken rats' striatum was developed to simultaneously quantitate three endogenous neurotransmitters, dopamine(DA), glutamate (Glu), and 5-hydroxytryptamine (5-HT), and nine components of Yuanhu-Baizhi herb pair in rats. The microdialysates were derivatizated with benzoyl chloride and analyzed on Agilent Poroshell 120EC-C18 column. The mobile phase consisted of aqueous solution with 0.05% formic acid and 2.0â¯mM ammonium formate (A), and acetonitrile (B) delivered at gradient elution. All the twelve analytes were quantified using an electrospray ionization tandem mass spectrometry in the selective reaction monitoring mode. The method was validated following FDA's guidance on bioanalytical validation. The standard curves were linear (â¯r2 > 0.991) over the corresponding concentration ranges. For three neurotransmitters, the intra-day and inter-day precision (RSD) were not greater than 14.9%, and the intra-day and inter-day accuracy (RE) ranged from -14.2â¼ 7.2%. The LLOQ of 5-HT, DA and Glu were 0.50â¯nM, 0.50â¯nM and 20.00â¯nM, respectively. For nine components in Yuanhu-Baizhi herb pair, the intra-day and inter-day precision (RSD) were less than 15.0%, and the intra-day and inter-day accuracy (RE) ranged from -12.5 â¼14.8%. The novel developed method was applied to pharmacokinetic (PK) and pharmacodynamic (PD) study of Yuanhu-Baizhi herb pair in rats. The results showed that the pharmacokinetic behavior of nine components in the herb pair was significantly different with that of single herb, mainly referring to the bioavailability of five alkaloids from Yuanhu increased whereas that of four coumarins decreased in rat striatum. Pharmacodynamic study based on Glu, DA, and 5-HT revealed that the content of neurotransmitters varied when rats were administered Yuanhu-Baizhi herb pair or single herb. PD analysis results demonstrated the compatibility of Yuanhu and Baizhi has the synergistic effect and the effect of attenuation from the point of the neurotransmitters' view. In conclusion, the pharmacokinetic and pharmacodynamic study based on in vivo microdialysis sampling from awaken animals and LC-MS/MS quantitation is in favor of revealing the compatibility mechanism of herb pairs.
Assuntos
Corpo Estriado/química , Medicamentos de Ervas Chinesas/farmacologia , Neurotransmissores/isolamento & purificação , Nociceptividade/efeitos dos fármacos , Angelica/química , Animais , Cromatografia Líquida de Alta Pressão/métodos , Estado de Consciência , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/fisiologia , Corydalis/química , Sinergismo Farmacológico , Quimioterapia Combinada , Medicamentos de Ervas Chinesas/química , Masculino , Microdiálise/métodos , Modelos Animais , Neurotransmissores/metabolismo , Nociceptividade/fisiologia , Ratos , Reprodutibilidade dos Testes , Rizoma/química , Espectrometria de Massas por Ionização por Electrospray , Técnicas Estereotáxicas , Espectrometria de Massas em Tandem/métodosRESUMO
Reading is a learned activity that engages multiple cognitive systems. In a cohort of typical and struggling adult readers we show evidence that successful oral reading of real words is related to gamma-amino-butyric acid (GABA) concentration in the higher-order language system, whereas reading of unfamiliar pseudo-words is not related to GABA in this system. We also demonstrate the capability of resting state functional connectivity (rsFC) combined with GABA measures to predict single real word compared to pseudo-word reading performance. Results show that the strength of rsFC between left fusiform gyrus (L-FG) and higher-order language systems predicts oral reading behavior of real words, irrespective of the local concentration of GABA. On the other hand, pseudo-words, which require grapheme-to-phoneme conversion, are not predicted by the connection between L-FG and higher-order language system. This suggests that L-FG may have a multi-functional role: lexical processing of real words and grapheme-to-phoneme processing of pseudo-words. Additionally, rsFC between L-FG, pre-motor, and putamen areas are positively related to the oral reading of both real and pseudo-words, suggesting that text may be converted into a phoneme sequence for speech initiation and production regardless of whether the stimulus is a real word or pseudo-word. In summary, from a systems neuroscience perspective, we show that: (i) strong rsFC between higher order visual, language, and pre-motor areas can predict and differentiate efficient oral reading of real and pseudo-words. (ii) GABA measures, along with rsFC, help to further differentiate the neural pathways for previously learned real words versus unfamiliar pseudo-words.
Assuntos
Corpo Estriado/fisiologia , Lobo Frontal/fisiologia , Aprendizagem/fisiologia , Leitura , Lobo Temporal/fisiologia , Ácido gama-Aminobutírico/metabolismo , Adulto , Mapeamento Encefálico , Corpo Estriado/química , Corpo Estriado/diagnóstico por imagem , Feminino , Lobo Frontal/química , Lobo Frontal/diagnóstico por imagem , Humanos , Linguística , Alfabetização , Imageamento por Ressonância Magnética , Espectroscopia de Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Vias Neurais/fisiologia , Lobo Temporal/química , Lobo Temporal/diagnóstico por imagem , Adulto Jovem , Ácido gama-Aminobutírico/análiseRESUMO
Striatal GABAergic interneurons that express nitric oxide synthase-so-called low-threshold spike interneurons (LTSIs)-play several key roles in the striatum. But what drives the activity of these interneurons is less well defined. To fill this gap, a combination of monosynaptic rabies virus mapping (msRVm), electrophysiological and optogenetic approaches were used in transgenic mice in which LTSIs expressed either Cre recombinase or a fluorescent reporter. The rabies virus studies revealed a striking similarity in the afferent connectomes of LTSIs and neighboring cholinergic interneurons, particularly regarding connections arising from the parafascicular nucleus of the thalamus and cingulate cortex. While optogenetic stimulation of cingulate inputs excited both cholinergic interneurons and LTSIs, thalamic stimulation excited cholinergic interneurons, but inhibited LTSIs. This inhibition was dependent on cholinergic interneurons and had two components: a previously described GABAergic element and one that was mediated by M4 muscarinic acetylcholine receptors. In addition to this phasic signal, cholinergic interneurons tonically excited LTSIs through a distinct, M1 muscarinic acetylcholine receptor pathway. This coordinated cholinergic modulation of LTSIs predisposed them to rhythmically burst in response to phasic thalamic activity, potentially reconfiguring striatal circuitry in response to salient environmental stimuli.
Assuntos
Neurônios Colinérgicos/metabolismo , Corpo Estriado/metabolismo , Giro do Cíngulo/metabolismo , Interneurônios/metabolismo , Óxido Nítrico/metabolismo , Tálamo/metabolismo , Animais , Neurônios Colinérgicos/química , Corpo Estriado/química , Feminino , Giro do Cíngulo/química , Interneurônios/química , Masculino , Camundongos , Camundongos Transgênicos , Óxido Nítrico/análise , Optogenética/métodos , Tálamo/químicaRESUMO
Parkinson's disease (PD) is characterized by the degeneration of dopaminergic neurons and excessive microglial activation in the substantia nigra pars compacta (SNpc). In the present study, we aimed to demonstrate the therapeutic effectiveness of the potent sphingosine-1-phosphate receptor antagonist fingolimod (FTY720) in an animal model of PD induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and to identify the potential mechanisms underlying these therapeutic effects. C57BL/6J mice were orally administered FTY720 before subcutaneous injection of MPTP. Open-field and rotarod tests were performed to determine the therapeutic effect of FTY720. The damage to dopaminergic neurons and the production of monoamine neurotransmitters were assessed using immunohistochemistry, high-performance liquid chromatography, and flow cytometry. Immunofluorescence (CD68- positive) and enzyme-linked immunosorbent assay were used to analyze the activation of microglia, and the levels of activated signaling molecules were measured using Western blotting. Our findings indicated that FTY720 significantly attenuated MPTP-induced behavioral deficits, reduced the loss of dopaminergic neurons, and increased dopamine release. FTY720 directly inhibited MPTP-induced microglial activation in the SNpc, suppressed the production of interleukin (IL)-6, IL-1ß, and tumor necrosis factor-α in BV-2 microglial cells treated with 1-methyl-4-phenylpyridinium (MPP+), and subsequently decreased apoptosis in SH-SY5Y neuroblastoma cells. Moreover, in MPP+-treated BV-2 cells and primary microglia, FTY720 treatment significantly attenuated the increases in the phosphorylation of PI3K/AKT/GSK-3ß, reduced ROS generation and p65 activation, and also inhibited the activation of NLRP3 inflammasome and caspase-1. In conclusion, FTY720 may reduce PD progression by inhibiting NLRP3 inflammasome activation via its effects on ROS generation and p65 activation in microglia. These findings provide novel insights into the mechanisms underlying the therapeutic effects of FTY720, suggesting its potential as a novel therapeutic strategy against PD. Graphical Abstract FTY720 may reduce ROS production by inhibiting the PI3K/AKT/GSK-3ß signaling pathway, while at the same time reducing p65 phosphorylation, thus decreasing NLRP3 inflammasome activation through these two pathways, ultimately reducing microglia activation-induced neuronal damage.
Assuntos
Antiparkinsonianos/farmacologia , Cloridrato de Fingolimode/farmacologia , Inflamassomos/efeitos dos fármacos , Microglia/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Transtornos Parkinsonianos/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Corpo Estriado/química , Corpo Estriado/efeitos dos fármacos , Citocinas/biossíntese , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/patologia , Avaliação Pré-Clínica de Medicamentos , Comportamento Exploratório/efeitos dos fármacos , Inflamassomos/metabolismo , Intoxicação por MPTP/tratamento farmacológico , Intoxicação por MPTP/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microglia/metabolismo , Mitocôndrias/efeitos dos fármacos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Transtornos Parkinsonianos/imunologia , Parte Compacta da Substância Negra/química , Parte Compacta da Substância Negra/efeitos dos fármacos , Parte Compacta da Substância Negra/patologia , Espécies Reativas de Oxigênio , Teste de Desempenho do Rota-Rod , Transdução de Sinais/efeitos dos fármacosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Kami-shoyo-san (KSS) is a Kampo formula used clinically for menopause-related symptoms in Japan. However, the effect of KSS on autism spectrum disorder (ASD), a developmental disorder with a higher prevalence in males than in females, has not been reported yet. AIM OF THE STUDY: It is accepted generally that dysfunction in the GABAergic system is associated with pathogenesis of ASD. In our previous study, a decrease in brain allopregnanolone (ALLO), a positive allosteric GABAA receptor modulator, induced ASD-like symptoms such as impaired sociability-related performance and increased repetitive self-grooming behavior in male mice, and that KSS ameliorated these behavioral abnormalities via GABAA receptor- and dopamine D1 receptor-mediated mechanisms. In this study, to better understand a gender difference in the prevalence of ASD, we examined whether dissection of ovary (OVX), a major organ secreting progesterone in females, causes ASD-like behaviors in a manner dependent on brain ALLO levels, and if so, how KSS affects the behaviors. MATERIALS AND METHODS: Six-week-old ICR female mice received ovariectomy, and KSS (74â¯mg/kg and 222â¯mg/kg, p.o.) were treated before 1â¯h starting each behavioral test. The sociability, social anxiety-like behavior, and self-grooming behavior were analyzed by the resident-intruder test, mirror chamber test, and open field test, respectively. After finishing the behavioral experiment, the ALLO content in the brain was measured by ELISA. Furthermore, we examined the effects of OVX on the neuro-signaling pathways in the prefrontal cortex and striatum by Western blotting. RESULTS: The results revealed that OVX induced sociability deficits and social anxiety-related behaviors, but not repetitive self-grooming behavior, and that these behavioral changes were accompanied not only by a decrease of brain ALLO levels, but also by impairment of CREB- and CaMKIIα-mediated neuro-signaling in the prefrontal cortex. Moreover, the administration of KSS had no effect on the brain ALLO level, but significantly ameliorated the OVX-induced behavioral and neurochemical changes via facilitation of GABAA receptor and dopamine D1 receptor-mediated neurotransmission. CONCLUSIONS: These findings suggest that a decrease in gonadal hormone-derived ALLO plays a major role in ASD-like behaviors in female mice and that KSS is beneficial for the treatment of ASD in females.
Assuntos
Transtorno do Espectro Autista/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Medicina Kampo/métodos , Comportamento Social , Animais , Transtorno do Espectro Autista/diagnóstico , Transtorno do Espectro Autista/psicologia , Técnicas de Observação do Comportamento , Comportamento Animal/efeitos dos fármacos , Corpo Estriado/química , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Medicamentos de Ervas Chinesas/uso terapêutico , Feminino , Masculino , Camundongos , Camundongos Endogâmicos ICR , Ovariectomia , Córtex Pré-Frontal/química , Córtex Pré-Frontal/efeitos dos fármacos , Córtex Pré-Frontal/metabolismo , Pregnanolona/análise , Pregnanolona/metabolismo , Receptores de Dopamina D1/metabolismo , Receptores de GABA-A/metabolismo , Fatores Sexuais , Resultado do TratamentoRESUMO
In vivo 1H magnetic resonance spectroscopy (1H-MRS) investigations of amyotrophic lateral sclerosis (ALS) mouse brain may provide neurochemical profiles and alterations in association with ALS disease progression. We aimed to longitudinally follow neurochemical evolutions of striatum, brainstem and motor cortex of mice transgenic for G93A mutant human superoxide dismutase type-1 (G93A-SOD1), an ALS model. Region-specific neurochemical alterations were detected in asymptomatic G93A-SOD1 mice, particularly in lactate (-19%) and glutamate (+8%) of brainstem, along with γ-amino-butyric acid (-30%), N-acetyl-aspartate (-5%) and ascorbate (+51%) of motor cortex. With disease progression towards the end-stage, increased numbers of metabolic changes of G93A-SOD1 mice were observed (e.g. glutamine levels increased in the brainstem (>+66%) and motor cortex (>+54%)). Through ALS disease progression, an overall increase of glutamine/glutamate in G93A-SOD1 mice was observed in the striatum (p < 0.01) and even more so in two motor neuron enriched regions, the brainstem and motor cortex (p < 0.0001). These 1H-MRS data underscore a pattern of neurochemical alterations that are specific to brain regions and to disease stages of the G93A-SOD1 mouse model. These neurochemical changes may contribute to early diagnosis and disease monitoring in ALS patients.
Assuntos
Esclerose Lateral Amiotrófica/metabolismo , Química Encefálica/fisiologia , Encéfalo/metabolismo , Mutação , Superóxido Dismutase/genética , Esclerose Lateral Amiotrófica/enzimologia , Animais , Ácido Ascórbico/análise , Ácido Aspártico/análogos & derivados , Ácido Aspártico/análise , Tronco Encefálico/química , Corpo Estriado/química , Modelos Animais de Doenças , Progressão da Doença , Ácido Glutâmico/análise , Glutamina/análise , Humanos , Ácido Láctico/análise , Espectroscopia de Ressonância Magnética , Camundongos , Camundongos Transgênicos , Córtex Motor/química , Ácido gama-Aminobutírico/análiseRESUMO
One of the main inputs driving striatal activity is the thalamostriatal projection. While the hypothesis postulating that the different thalamostriatal projections contribute differentially to shape the functions of the striatum is largely accepted, existing technical limitations have hampered efforts to prove it. Here, through the use of electrophysiological recordings of antidromically photo-identified thalamostriatal neurons and the optogenetic inhibition of thalamostriatal terminals, we identify that the thalamostriatal projections from the parafascicular and the ventroposterior regions of the thalamus contribute to the smooth initiation and the appropriate execution of a sequence of movements. Our results support a model in which both thalamostriatal projections have specific contributions to the initiation and execution of sequences, highlighting the specific contribution of the ventroposterior thalamostriatal connection for the repetition of actions.
Assuntos
Corpo Estriado/fisiologia , Intenção , Movimento/fisiologia , Rede Nervosa/fisiologia , Tálamo/fisiologia , Animais , Corpo Estriado/química , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Rede Nervosa/química , Optogenética/métodos , Tálamo/químicaRESUMO
Royal Jelly (RJ) is a unique substance obtained from bees that has been used widely in European and Asian traditional medicine for its potential to prevent signs of aging through its antioxidative, anti-inflammatory, anti-hyperglycemic and anti-hypercholesterolemic properties. We recently reported an enhancement in spatial memory along with changes in monoaminergic transmission in aged rats after chronic RJ administration. Here, we aim to further explore the action of RJ on central nervous system activity by examining levels of amino acids in selected brain structures of aged male Wistar rats following 2-months of Greek RJ administration. RJ powder was previously chemically characterized and given orally (50 or 100â¯mg of powder/kg b.w./day) by gastric gavage. The concentrations of amino acids (alanine, aspartic acid, gamma-aminobutyric acid, glutamic acid, histidine and taurine) in the brain regions examined (prefrontal cortex, hippocampus, striatum and hypothalamus) were quantified using HPLC. We also examined basic biochemical parameters of renal and hepatic activity, as damage of these organs could potentially explain the changes in brain function and behavior. Upon biochemical examination, a decrease in the concentration of gamma-aminobutyric acid was observed in both the striatum and hypothalamus. Liver and kidney functions were not changed by chronic RJ-administration. Our results provide insight toward understanding the mechanism of action of RJ and its effects on neurotransmission in the central nervous system.
Assuntos
Corpo Estriado/efeitos dos fármacos , Ácidos Graxos/administração & dosagem , Hipotálamo/efeitos dos fármacos , Ácido gama-Aminobutírico/análise , Aminoácidos/análise , Animais , Corpo Estriado/química , Hipocampo/química , Hipocampo/efeitos dos fármacos , Hipotálamo/química , Masculino , Córtex Pré-Frontal/química , Córtex Pré-Frontal/efeitos dos fármacos , Ratos WistarRESUMO
Opiates, one of the oldest known drugs, are the benchmark for treating pain. Regular opioid exposure also induces euphoria making these compounds addictive and often misused, as shown by the current epidemic of opioid abuse and overdose mortalities. In addition to the effect of opioids on their cognate receptors and signaling cascades, these compounds also induce multiple adaptations at cellular and behavioral levels. As omega-3 polyunsaturated fatty acids (n-3 PUFAs) play a ubiquitous role in behavioral and cellular processes, we proposed that supplemental n-3 PUFAs, enriched in docosahexanoic acid (DHA), could offset these adaptations following chronic opioid exposure. We used an 8 week regimen of n-3 PUFA supplementation followed by 8 days of morphine in the presence of this diet. We first assessed the effect of morphine in different behavioral measures and found that morphine increased anxiety and reduced wheel-running behavior. These effects were reduced by dietary n-3 PUFAs without affecting morphine-induced analgesia or hyperlocomotion, known effects of this opiate acting at mu opioid receptors. At the cellular level we found that morphine reduced striatal DHA content and that this was reversed by supplemental n-3 PUFAs. Chronic morphine also increased glutamatergic plasticity and the proportion of Grin2B-NMDARs in striatal projection neurons. This effect was similarly reversed by supplemental n-3 PUFAs. Gene analysis showed that supplemental PUFAs offset the effect of morphine on genes found in neurons of the dopamine receptor 2 (D2)-enriched indirect pathway but not of genes found in dopamine receptor 1(D1)-enriched direct-pathway neurons. Analysis of the D2 striatal connectome by a retrogradely transported pseudorabies virus showed that n-3 PUFA supplementation reversed the effect of chronic morphine on the innervation of D2 neurons by the dorsomedial prefontal and piriform cortices. Together these changes outline specific behavioral and cellular effects of morphine that can be reduced or reversed by dietary n-3 PUFAs.
Assuntos
Ácidos Graxos Ômega-3/farmacologia , Morfina/farmacologia , Animais , Ansiedade/induzido quimicamente , Corpo Estriado/química , Esquema de Medicação , Feminino , Lobo Frontal/química , Lipídeos/análise , Locomoção/efeitos dos fármacos , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Morfina/administração & dosagem , Morfina/antagonistas & inibidores , Atividade Motora/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Glutamato/análiseRESUMO
Silica nanoparticles (SiNPs) are being used increasingly in biomedical and industrial fields; however, their adverse effects on human health have not been fully investigated. In this study, we focused on some of the toxicological aspects of SiNPs by studying oxidative stress and pro-inflammatory responses in the frontal cortex, corpus striatum and hippocampus regions of rat brain. Wistar rats were exposed to SiNPs of size 80 nm and 10 nm at a dose of 150 µg/50 µL phosphate-buffered saline/rat for 30 days. The results indicated a significant increase of lipid peroxide levels and hydrogen peroxide content in various regions of the treated rat brain. Moreover, these changes were accompanied with a significant decrease in the activities of manganese superoxide dismutase, glutathione reductase, catalase and reduced glutathione in different brain regions, suggesting impaired antioxidant defence system. Furthermore, SiNPs exposure not only increased messenger RNA (mRNA) and protein expression of nuclear factor-κB (NF-κB) but also significantly increased the mRNA and protein levels of tumour necrosis factor α (TNF-α), interleukin 1ß (IL-1ß) and monocyte chemoattractant protein 1 (MCP-1) in different regions of rat brain. Cumulatively, these data suggest that SiNPs induced the activation of NF-κB and increased the expression of TNF-α, IL-1ß and MCP-1 in rat brain, possibly via redox-sensitive cellular signalling pathways.
Assuntos
Encéfalo/efeitos dos fármacos , Nanopartículas/efeitos adversos , Dióxido de Silício/efeitos adversos , Administração Intranasal , Animais , Corpo Estriado/química , Corpo Estriado/efeitos dos fármacos , Lobo Frontal/química , Lobo Frontal/efeitos dos fármacos , Hipocampo/química , Hipocampo/efeitos dos fármacos , Peróxido de Hidrogênio/análise , Inflamação/induzido quimicamente , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Nanopartículas/administração & dosagem , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Dióxido de Silício/administração & dosagem , Superóxido Dismutase/metabolismoRESUMO
Recent studies have shown that Korean Red Ginseng (KRG) suppresses dopaminergic neuronal death in the brain of a Parkinson's disease (PD) mouse model, but the mechanism is still elusive. Using a 2-dimensional electrophoresis technique, we investigated whether KRG can restore the changes in protein expressions in the striatum (ST) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-injected mice. Male C57BL/6 mice (9 weeks old) were injected with 20 mg/kg MPTP intraperitoneally four times at 2-h intervals. KRG (100 mg/kg) was orally administered once a day for 3 days from one hour after the first MPTP injection. Two hours after the third KRG administration a pole test was performed to evaluate motor function, after which the brains were immediately harvested. Survival of dopaminergic neurons in the nigrostriatal pathway and protein expression in the ST were measured by immunohistochemistry and 2-dimensional electrophoresis. KRG suppressed MPTP-induced behavioral dysfunction and neuronal death in the nigrostriatal pathway. Moreover, 30 proteins changed by MPTP and KRG in the ST were identified and shown to be related to glycolysis/gluconeogenesis and neurodegenerative diseases including Alzheimer's disease and PD. KRG has neuroprotective effects against MPTP toxicity and alleviates protein expression profiles related to enhancing energy metabolism in the ST of MPTP-treated mice.
Assuntos
Corpo Estriado/efeitos dos fármacos , Fármacos Neuroprotetores/uso terapêutico , Panax , Transtornos Parkinsonianos/tratamento farmacológico , Fitoterapia/métodos , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/farmacologia , Animais , Western Blotting , Corpo Estriado/química , Modelos Animais de Doenças , Eletroforese em Gel Bidimensional , Masculino , Camundongos , Atividade Motora/efeitos dos fármacos , Proteínas do Tecido Nervoso/análise , Panax/química , ProteômicaRESUMO
Neurodegeneration may be involved in the development of tardive dyskinesia (TD), and low levels of brain-derived neurotrophic factor (BDNF) may play a role. Ginkgo biloba (EGb761), a potent antioxidant, may have neuroprotective effects. We hypothesized that there would be decreased BDNF expression in TD, but that treatment with EGb761 would increase BDNF expression and reduce TD manifestations in a rat model. Forty rats were treated with haloperidol (2mg/kg/day via intraperitoneal injections) for 5weeks. EGb761 (50mg/kg/day) and vitamin E (20mg/kg/day) were then administered by oral gavage for another 5weeks, and we compared the effects of treatment with EGb761 or vitamin E on haloperidol-induced vacuous chewing movements (VCMs) and BDNF expression in four brain regions: prefrontal cortex (PFC), striatum (ST), substantia nigra (SNR), and globus pallidus (GP). Our results showed that haloperidol administration led to a progressive increase in VCMs, but both EGb761 and vitamin E significantly decreased VCMs. Haloperidol also decreased BDNF expression in all four brain regions, but both EGb761 and vitamin E administration significantly increased BDNF expression. Our results showed that both EGb761 and VE treatments exerted similar positive effects in a rat model of TD and increased BDNF expression levels in the four tested brain regions, suggesting that both EGb761 and vitamin E improve TD symptoms, possibly by enhancing BDNF in the brain and/or via their free radical-scavenging actions.
Assuntos
Antioxidantes/farmacologia , Fator Neurotrófico Derivado do Encéfalo/análise , Haloperidol/farmacologia , Mastigação/efeitos dos fármacos , Extratos Vegetais/farmacologia , Discinesia Tardia/tratamento farmacológico , Vitamina E/farmacologia , Animais , Corpo Estriado/química , Modelos Animais de Doenças , Ginkgo biloba , Masculino , Extratos Vegetais/uso terapêutico , Ratos , Ratos Sprague-Dawley , Substância Negra/química , Discinesia Tardia/metabolismo , Vitamina E/uso terapêuticoRESUMO
Various iron-oxide nanoparticles have been in use for a long time as therapeutic and imaging agents and for supplemental delivery in cases of iron-deficiency. While all of these products have a specified size range of â¼ 40 nm and above, efforts are underway to produce smaller particles, down to â¼ 1 nm. Here, we show that after a 24-h exposure of SHSY-5Y human neuroblastoma cells to 10 µg/ml of 10 and 30 nm ferric oxide nanoparticles (Fe-NPs), cellular dopamine content was depleted by 68 and 52 %, respectively. Increases in activated tyrosine kinase c-Abl, a molecular switch induced by oxidative stress, and neuronal α-synuclein expression, a protein marker associated with neuronal injury, were also observed (55 and 38 % percent increases, respectively). Inhibition of cell-proliferation, significant reductions in the number of active mitochondria, and a dose-dependent increase in reactive oxygen species (ROS) were observed in neuronal cells. Additionally, using a rat in vitro blood-brain barrier (BBB) model, a dose-dependent increase in ROS accompanied by increased fluorescein efflux demonstrated compromised BBB integrity. To assess translational implications, in vivo Fe-NP-induced neurotoxicity was determined using in vivo MRI and post-mortem neurochemical and neuropathological correlates in adult male rats after exposure to 50 mg/kg of 10 nm Fe-NPs. Significant decrease in T 2 values was observed. Dynamic observations suggested transfer and retention of Fe-NPs from brain vasculature into brain ventricles. A significant decrease in striatal dopamine and its metabolites was also observed, and neuropathological correlates provided additional evidence of significant nerve cell body and dopaminergic terminal damage as well as damage to neuronal vasculature after exposure to 10 nm Fe-NPs. These data demonstrate a neurotoxic potential of very small size iron nanoparticles and suggest that use of these ferric oxide nanoparticles may result in neurotoxicity, thereby limiting their clinical application.
Assuntos
Neurônios Dopaminérgicos/efeitos dos fármacos , Nanopartículas de Magnetita/toxicidade , Animais , Apoptose/efeitos dos fármacos , Barreira Hematoencefálica/efeitos dos fármacos , Caspases/metabolismo , Catecolaminas/análise , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Corpo Estriado/química , Corpo Estriado/efeitos dos fármacos , Neurônios Dopaminérgicos/química , Neurônios Dopaminérgicos/ultraestrutura , Ativação Enzimática/efeitos dos fármacos , Humanos , Imageamento por Ressonância Magnética , Espectroscopia de Ressonância Magnética , Masculino , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Nanosferas , Neuroblastoma/patologia , Estresse Oxidativo , Tamanho da Partícula , Permeabilidade/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/análise , Espectrometria por Raios XRESUMO
Parkinson's disease (PD) is a neurodegenerative disorder characterized by the irreversible loss of dopaminergic neurons in the nigrostriatal pathway with subsequent dopamine deficiency. Environmental causes have been proposed through molecules, such as 1-methyl-4-phenylpyridinium (MPP(+)), to induce oxidative stress. The methanolic extract of plants of the genus Buddleja has been reported to have in vitro and in vivo antioxidant properties to protect against neuronal death. In the present study, the neuroprotective effect of Buddleja cordata methanolic extract in the MPP(+) PD rat model was investigated. Animals were administered orally with 50 or 100 mg/kg of methanolic extract every 24 h for 14 days. Twenty hours later, rats were infused with an intrastriatal stereotaxic microinjection of 10 µg MPP(+) in 8 µl sterile saline solution. Six days later, the animals were treated with 1 mg/kg apomorphine to record ipsilateral rotations for 1 h. All the rats were killed by decapitation and the lesioned striatum was dissected for dopamine and lipid peroxidation quantifications. Both methanolic extract doses led to a significantly lower (P < 0.05) number of ipsilateral rotations (75-80 %). This behavioral protection was corroborated with 60 % level of dopamine preservation (P < 0.05) and 90 % decrease in the formation of lipidic fluorescent products in the striatum (P < 0.05). This study demonstrates the antioxidant and neuroprotective effect of Buddleja cordata methanolic extract in the MPP(+) PD rat model, possibly due to the involvement of phenylpropanoids.
Assuntos
Antioxidantes/uso terapêutico , Buddleja , Intoxicação por MPTP/tratamento farmacológico , Fármacos Neuroprotetores/uso terapêutico , Fitoterapia , Animais , Corpo Estriado/química , Dopamina/análise , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Metanol , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/uso terapêutico , Ratos WistarRESUMO
Ferulic acid is a polyphenol that has antioxidant, anti-inflammatory and anticancer properties. The present study analyzed the antidepressant-like potential of ferulic acid using two well-validated mouse models of despair test, tail suspension and forced swim tests. The results suggested that ferulic acid treatment at doses of 10, 20, 40 and 80 mg/kg (p.o.) significantly reduced the immobility time in both of these two tests. These doses that affected the depressive-like behaviors did now show any effect on locomotion counts. The further neurochemical assays suggested that ferulic acid increased monoamine neurotransmitter levels in the brain regions that are relative to mood disorders: the hippocampus and frontal cortex. The increased tend to serotonin and norepinephrine was also found in the hypothalamus after higher dose of ferulic acid treatment. The subsequent study suggested that monoamine oxidase A (MAO-A) activity was inhibited in the frontal cortex and hippocampus when treatment with 40 and 80 mg/kg ferulic acid; while MAO-B activity did not change significantly. The current study provides the first lines of evidence that serotonin and norepinephrine, but not dopamine levels were elevated in mouse hippocampus and frontal cortex after ferulic acid treatment. These changes may be attributable to the inhibition of MAO-A activities in the same brain regions.
Assuntos
Antidepressivos/uso terapêutico , Ácidos Cumáricos/uso terapêutico , Inibidores da Monoaminoxidase/uso terapêutico , Norepinefrina/fisiologia , Serotonina/fisiologia , Animais , Antidepressivos/farmacologia , Corpo Estriado/química , Corpo Estriado/efeitos dos fármacos , Ácidos Cumáricos/farmacologia , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Lobo Frontal/química , Lobo Frontal/efeitos dos fármacos , Hipocampo/química , Hipocampo/efeitos dos fármacos , Hipotálamo/química , Hipotálamo/efeitos dos fármacos , Imipramina/farmacologia , Imipramina/uso terapêutico , Imobilização , Masculino , Camundongos , Camundongos Endogâmicos ICR , Moclobemida/farmacologia , Moclobemida/uso terapêutico , Monoaminoxidase/análise , Inibidores da Monoaminoxidase/farmacologia , Atividade Motora/efeitos dos fármacos , Norepinefrina/análise , Esforço Físico/efeitos dos fármacos , Serotonina/análise , Estresse Fisiológico , Estresse Psicológico , NataçãoRESUMO
Drugs that selectively activate estrogen receptor ß (ERß) are potentially safer than the nonselective estrogens currently used in hormonal replacement treatments that activate both ERß and ERα. The selective ERß agonist AC-186 was evaluated in a rat model of Parkinson's disease induced through bilateral 6-hydroxydopamine lesions of the substantia nigra. In this model, AC-186 prevented motor, cognitive, and sensorimotor gating deficits and mitigated the loss of dopamine neurons in the substantia nigra, in males, but not in females. Furthermore, in male rats, 17ß-estradiol, which activates ERß and ERα with equal potency, did not show the same neuroprotective benefits as AC-186. Hence, in addition to a beneficial safety profile for use in both males and females, a selective ERß agonist has a differentiated pharmacological profile compared to 17ß-estradiol in males.
Assuntos
Cicloexanos/uso terapêutico , Receptor beta de Estrogênio/agonistas , Fármacos Neuroprotetores/uso terapêutico , Transtornos Parkinsonianos/tratamento farmacológico , Fenóis/uso terapêutico , Animais , Corpo Estriado/química , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/patologia , Cicloexanos/química , Cicloexanos/farmacologia , Citocinas/análise , Avaliação Pré-Clínica de Medicamentos , Estradiol/uso terapêutico , Comportamento Exploratório/efeitos dos fármacos , Feminino , Masculino , Estrutura Molecular , Proteínas do Tecido Nervoso/análise , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/farmacologia , Ovariectomia , Oxidopamina/toxicidade , Transtornos Parkinsonianos/induzido quimicamente , Fenóis/química , Fenóis/farmacologia , Ratos , Ratos Sprague-Dawley , Reflexo de Sobressalto/efeitos dos fármacos , Teste de Desempenho do Rota-Rod , Fatores Sexuais , Substância Negra/química , Substância Negra/efeitos dos fármacos , Substância Negra/patologia , Tirosina 3-Mono-Oxigenase/análiseRESUMO
We report results of studies of global and targeted neuronal metabolomes by ambient pressure ion mobility mass spectrometry. The rat frontal cortex, striatum, and thalamus were sampled from control nontreated rats and those treated with acute cocaine or pargyline. Quantitative evaluations were made by standard additions or isotopic dilution. The mass detection limit was ~100 pmol varying with the analyte. Targeted metabolites of dopamine, serotonin, and glucose followed the rank order of distribution expected between the anatomical areas. Data was evaluated by principal component analysis on 764 common metabolites (identified by m/z and reduced mobility). Differences between anatomical areas and treatment groups were observed for 53 % of these metabolites using principal component analysis. Global and targeted metabolic differences were observed between the three anatomical areas with contralateral differences between some areas. Following drug treatments, global and targeted metabolomes were found to shift relative to controls and still maintained anatomical differences. Pargyline reduced 3,4-dihydroxyphenylacetic acid below detection limits, and 5-HIAA varied between anatomical regions. Notable findings were: (1) global metabolomes were different between anatomical areas and were altered by acute cocaine providing a broad but targeted window of discovery for metabolic changes produced by drugs of abuse; (2) quantitative analysis was demonstrated using isotope dilution and standard addition; (3) cocaine changed glucose and biogenic amine metabolism in the anatomical areas tested; and (4) the largest effect of cocaine was on the glycolysis metabolome in the thalamus confirming inferences from previous positron emission tomography studies using 2-deoxyglucose.
Assuntos
Cocaína/farmacologia , Corpo Estriado/efeitos dos fármacos , Dopamina/metabolismo , Lobo Frontal/efeitos dos fármacos , Glucose/metabolismo , Serotonina/metabolismo , Tálamo/efeitos dos fármacos , Ácido 3,4-Di-Hidroxifenilacético/análise , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Animais , Corpo Estriado/química , Corpo Estriado/metabolismo , Dopamina/análise , Lobo Frontal/química , Lobo Frontal/metabolismo , Glucose/análise , Glicólise/efeitos dos fármacos , Ácido Hidroxi-Indolacético/análise , Ácido Hidroxi-Indolacético/metabolismo , Limite de Detecção , Masculino , Espectrometria de Massas , Metabolômica , Pargilina/farmacologia , Análise de Componente Principal , Ratos , Ratos Sprague-Dawley , Serotonina/análise , Tálamo/química , Tálamo/metabolismoRESUMO
The serotonergic system and the hypothalamic-pituitary-adrenal (HPA) axis are crucially involved in the regulation of emotions. Specifically, spontaneous and/or environmentally mediated modulations of the functionality of these systems early in development may favour the onset of depressive- and anxiety-related phenotypes. While the independent contribution of each of these systems to the emergence of abnormal phenotypes has been detailed in clinical and experimental studies, only rarely has their interaction been systematically investigated. Here, we addressed the effects of reduced serotonin and environmental stress during the early stages of postnatal life on emotional regulations in mice. To this aim, we administered, to outbred CD1 mouse dams, during their first week of lactation, a tryptophan deficient diet (T) and corticosterone via drinking water (C; 80µg/ml). Four groups of dams (animal facility rearing, AFR; T treated, T; C treated, C; T and C treated, TC) and their male offspring were used in the study. Maternal care was scored throughout treatment and adult offspring were tested for: anhedonia (progressive ratio schedule); anxiety-related behaviour (approach-avoidance conflict paradigm); BDNF, dopamine and serotonin concentrations in selected brain areas. T, C and TC treatments reduced active maternal care compared to AFR. Adult TC offspring showed significantly increased anxiety- and anhedonia-related behaviours, reduced striatal and increased hypothalamic BDNF and reduced dopamine and serotonin in the prefrontal cortex and their turnover in the hippocampus. Thus, present findings support the view that neonatal variations in the functionality of the serotonergic system and of HPA axis may jointly contribute to induce emotional disturbances in adulthood.
Assuntos
Anedonia/fisiologia , Ansiedade/etiologia , Corticosterona/toxicidade , Emoções/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/fisiopatologia , Sistema Hipófise-Suprarrenal/fisiopatologia , Serotonina/fisiologia , Triptofano/deficiência , Anedonia/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Animais Lactentes , Ansiedade/induzido quimicamente , Ansiedade/fisiopatologia , Fator Neurotrófico Derivado do Encéfalo/análise , Corpo Estriado/química , Dopamina/análise , Emoções/fisiologia , Comportamento Exploratório/efeitos dos fármacos , Comportamento Exploratório/fisiologia , Feminino , Sistema Hipotálamo-Hipofisário/química , Hipotálamo/química , Masculino , Comportamento Materno/efeitos dos fármacos , Comportamento Materno/fisiologia , Camundongos , Serotonina/análise , Serotonina/deficiênciaRESUMO
We examined thalamic input to striatum in rats using immunolabeling for the vesicular glutamate transporter (VGLUT2). Double immunofluorescence viewed with confocal laser scanning microscopy (CLSM) revealed that VGLUT2+ terminals are distinct from VGLUT1+ terminals. CLSM of Phaseolus vulgaris-leucoagglutinin (PHAL)-labeled cortical or thalamic terminals revealed that VGLUT2 is rare in corticostriatal terminals but nearly always present in thalamostriatal terminals. Electron microscopy revealed that VGLUT2+ terminals made up 39.4% of excitatory terminals in striatum (with VGLUT1+ corticostriatal terminals constituting the rest), and 66.8% of VGLUT2+ terminals synapsed on spines and the remainder on dendrites. VGLUT2+ axospinous terminals had a mean diameter of 0.624 µm, while VGLUT2+ axodendritic terminals a mean diameter of 0.698 µm. In tissue in which we simultaneously immunolabeled thalamostriatal terminals for VGLUT2 and striatal neurons for D1 (with about half of spines immunolabeled for D1), 54.6% of VGLUT2+ terminals targeted D1+ spines (i.e., direct pathway striatal neurons), and 37.3% of D1+ spines received VGLUT2+ synaptic contacts. By contrast, 45.4% of VGLUT2+ terminals targeted D1-negative spines (i.e., indirect pathway striatal neurons), and only 25.8% of D1-negative spines received VGLUT2+ synaptic contacts. Similarly, among VGLUT2+ axodendritic synaptic terminals, 59.1% contacted D1+ dendrites, and 40.9% contacted D1-negative dendrites. VGLUT2+ terminals on D1+ spines and dendrites tended to be slightly smaller than those on D1-negative spines and dendrites. Thus, thalamostriatal terminals contact both direct and indirect pathway striatal neurons, with a slight preference for direct. These results are consistent with physiological studies indicating slightly different effects of thalamic input on the two types of striatal projection neurons.
Assuntos
Corpo Estriado/ultraestrutura , Neurônios/ultraestrutura , Tálamo/ultraestrutura , Proteína Vesicular 2 de Transporte de Glutamato/ultraestrutura , Animais , Corpo Estriado/química , Corpo Estriado/citologia , Potenciais Pós-Sinápticos Excitadores/fisiologia , Masculino , Microscopia Confocal/métodos , Vias Neurais/química , Vias Neurais/citologia , Vias Neurais/ultraestrutura , Neurônios/química , Ratos , Ratos Sprague-Dawley , Tálamo/química , Tálamo/citologia , Proteína Vesicular 2 de Transporte de Glutamato/fisiologiaRESUMO
OBJECTIVE: Ningdong granule is a traditional Chinese medicine preparation for the treatment of Tourette's syndrome. METHODS: Sixty-four rats were randomly assigned to a control group and three experimental groups, respectively. Rat models of Tourette's syndrome were established via intraperitoneal injection of apomorphine (Apo). The rats in the experimental groups were subsequently intragastrically injected with haloperidol at 10 mg/kg (haloperidol group), ningdong granule at 370 mg/kg (NDG group), and normal saline (0.9%) at 10 mL/kg (Apo group), respectively. Rat behaviors were observed and recorded on a daily basis. After 12 w, all rats were sacrificed, and sera and striatal tissues were harvested. Homovanillic acid levels in sera, as well as dopamine and dopamine D2 receptor mRNA expression in the striatum, were measured to determine possible mechanisms of Ningdong granule on the dopamine system in a rat model ofTourette's syndrome. RESULTS: Following intervention, stereotype actions of the Tourette's syndrome rats were significantly inhibited in the haloperidol and NDG groups, respectively (P < 0.01). Homovanillic levels were significantly greater in the haloperidol and NDG groups, respectively (P < 0.05). In addition, dopamine levels were significantly less in the NDG group (P < 0.01), and DRD2 mRNA expression was significantly reduced in the haloperidol and NDG groups, respectively (P < 0.05). CONCLUSION: Results demonstrated that Ning-dong granule effectively inhibited stereotype actions and Tourette's syndrome symptoms by promoting dopamine metabolism, reducing dopamine levels in the striatum, increasing homovanillic acid content in sera, and reducing mRNA expression of DRD2 in the striatum.