Dyskinesias or abnormal involuntary movements are a debilitating complication of long-term levodopa treatment of Parkinson’s disease (PD) that is widely experienced and may compromise the efficacy of the drug therapy. Tardive dyskinesia is another important adverse effect seen with antipsychotic drug treatment. The neural mechanisms underlying levodopa- and neuroleptic-induced dyskinesia are not clear and involvement of the endogenous opioid neuropeptide system has been implicated. In this thesis, the role of the opioid system is investigated in models of dyskinesia and PD using behavioral, neurochemical and advanced analytical chemistry techniques. In addition, the motor effects of a new partial dopamine agonist with normalizing properties on both reduced and elevated dopamine transmission are studied and a new model for tardive dyskinesia is presented. Using microdialysis in combination with micro-electrospray mass spectrometry, the in vivo processing of the opioid neuropeptide dynorphin A(1-17) was studied and 32 metabolites were detected in the striatum. Altered in vivo metabolism of the peptide was found in a model of PD with more metabolites formed in the dopamine-depleted striatum. Moreover, dynorphin A(1-17) was differently processed in levodopa-, bromocriptine and saline-treated animals. Levodopa treatment caused an increase in the mRNA expression of the precursor of dynorphin, preproenkephalin-B as well as the precursor of enkephalin, preproenkephalin-A, in all sub-regions of the dopamine-depleted striatum. A non-selective opioid receptor antagonist, naloxone, was found to reduce levodopa-induced dyskinesia with maintained antiparkinsonian response and a normalization of hyperkinesia. Moreover, the new drug GMC1111 showed dopamine stabilizing properties in models of levodopa-induced dyskinesia and PD. This might prove useful in the treatment of PD. Altogether, these results suggest that the endogenous opioid system is involved in the pathophysiology of levodopa-induced dyskinesia.

No description available.

Dopamine deficiency in the striatum is a central feature of Parkinson’s disease (PD). Symptomatic therapy with L-3,4-dihydroxyphenylalanine (L-DOPA) aims at restoring physiological dopaminergic neurotransmission within the brain. Unfortunately, current treatment paradigms fail to achieve this goal, which leads to the emergence of motor complications, secondary to long term L-DOPA administration, including dyskinesia and wearing-OFF, and non-motor symptoms related to disease progression, including neuropsychiatric symptoms such as psychosis. However, degenerative changes in PD are not limited to the dopaminergic system, but also affect the serotonergic system. There is increasing evidence suggesting an involvement of the serotonergic system in the pathophysiology of both motor and non-motor complications of PD. The work presented in this Thesis has investigated the serotonergic and dopaminergic systems in PD, by performing post mortem studies in the brains of PD patients and of parkinsonian non-human primates (NHPs), and by performing behavioural studies in the parkinsonian rat and NHP models of PD. The main conclusions presented are that: 1) serotonergic type 1A (5-HT1A) and 2A (5-HT2A) levels are altered in the brains of dyskinetic parkinsonian NHPs, suggesting abnormal 5-HT1A- and 5-HT2A-mediated neurotransmission in dyskinesia; 2) 5-HT2A receptor levels are altered in the brains of PD patients with visual hallucinations (VH), suggesting abnormal 5-HT2A-mediated neurotransmission in VH; 3) some of the anti-dyskinetic actions attributed to stimulating 5-HT1A or antagonising 5-HT2A receptors might in fact be due to an antagonist action at D4 receptors, as antagonising D4 receptors significantly alleviates L-DOPA-induced dyskinesia in rat and NHP models of PD; 4) concurrent inhibition of the serotonin and dopamine transporters (SERT and DAT, respectively) enhances duration of L-DOPA-induced ON-time in the parkinsonian NHP. However, the ratio of SERT/ DAT inhibition appears crucial in determining the quality of this extra ON-time; SERT > DAT inhibition exacerbates the severity of L-DOPA-induced dyskinesia, whereas SERT = DAT and DAT > SERT inhibition do not worsen the severity of L-DOPA-induced dyskinesia. Together these data extend our knowledge of the interaction between serotonin and dopamine, specifically as they relate to symptoms and side effects of dopamine replacement therapy in PD and highlight potential novel therapeutic approaches to PD.

Dopamine deficiency in the striatum is a central feature of Parkinson’s disease (PD). Symptomatic therapy with L-3,4-dihydroxyphenylalanine (L-DOPA) aims at restoring physiological dopaminergic neurotransmission within the brain. Unfortunately, current treatment paradigms fail to achieve this goal, which leads to the emergence of motor complications, secondary to long term L-DOPA administration, including dyskinesia and wearing-OFF, and non-motor symptoms related to disease progression, including neuropsychiatric symptoms such as psychosis. However, degenerative changes in PD are not limited to the dopaminergic system, but also affect the serotonergic system. There is increasing evidence suggesting an involvement of the serotonergic system in the pathophysiology of both motor and non-motor complications of PD. The work presented in this Thesis has investigated the serotonergic and dopaminergic systems in PD, by performing post mortem studies in the brains of PD patients and of parkinsonian non-human primates (NHPs), and by performing behavioural studies in the parkinsonian rat and NHP models of PD. The main conclusions presented are that: 1) serotonergic type 1A (5-HT1A) and 2A (5-HT2A) levels are altered in the brains of dyskinetic parkinsonian NHPs, suggesting abnormal 5-HT1A- and 5-HT2A-mediated neurotransmission in dyskinesia; 2) 5-HT2A receptor levels are altered in the brains of PD patients with visual hallucinations (VH), suggesting abnormal 5-HT2A-mediated neurotransmission in VH; 3) some of the anti-dyskinetic actions attributed to stimulating 5-HT1A or antagonising 5-HT2A receptors might in fact be due to an antagonist action at D4 receptors, as antagonising D4 receptors significantly alleviates L-DOPA-induced dyskinesia in rat and NHP models of PD; 4) concurrent inhibition of the serotonin and dopamine transporters (SERT and DAT, respectively) enhances duration of L-DOPA-induced ON-time in the parkinsonian NHP. However, the ratio of SERT/ DAT inhibition appears crucial in determining the quality of this extra ON-time; SERT > DAT inhibition exacerbates the severity of L-DOPA-induced dyskinesia, whereas SERT = DAT and DAT > SERT inhibition do not worsen the severity of L-DOPA-induced dyskinesia. Together these data extend our knowledge of the interaction between serotonin and dopamine, specifically as they relate to symptoms and side effects of dopamine replacement therapy in PD and highlight potential novel therapeutic approaches to PD.

Certains neuropeptides (enképhaline et neurotensine) sont des modulateurs du système dopaminergique. Chez les rongeurs, le traitement avec l’antipsychotique typique halopéridol (antagoniste des récepteurs D2), augmente fortement leurs niveaux d’ARNm dans le striatum, une structure centrale du système dopaminergique qui contrôle l’activité locomotrice. Comme l’halopéridol est associé avec de nombreux effets secondaires moteurs, on peut penser que la modulation des neuropeptides est possiblement un mécanisme d’adaptation visant à rétablir l’homéostasie du système dopaminergique après le blocage des récepteurs D2. Cependant, le mécanisme moléculaire de cette régulation transcriptionnelle n’est pas bien compris. Nur77 est un facteur de transcription de la famille des récepteurs nucléaires orphelins qui agit en tant que gène d’induction précoce. Le niveau de son ARNm est aussi fortement augmenté dans le striatum suivant un traitement avec halopéridol. Plusieurs évidences nous suggèrent que Nur77 est impliqué dans la modulation transcriptionnelle des neuropeptides. Nur77 peut former des hétérodimères fonctionnels avec le récepteur rétinoïde X (RXR). En accord avec une activité transcriptionnelle d’un complexe Nur77/RXR, l’agoniste RXR (DHA) réduit tandis que l’antagoniste RXR (HX531) augmente les troubles moteurs induits par un traitement chronique à l’halopéridol chez les souris sauvages tandis que ces ligands pour RXR n’ont aucun effet chez les souris Nur77 nulles. Nos travaux ont révélé que l’antagoniste RXR (HX531) réduit l’augmentation des niveaux d’enképhaline suivant un traitement chronique avec l’halopéridol. Nous avons ensuite démontré la liaison in vitro de Nur77 sur un élément de réponse présent dans le promoteur proximal de la proenképhaline, le peptide précurseur de l’enképhaline. Ces résultats supportent l’hypothèse que Nur77, en combinaison avec RXR, pourrait participer à la régulation transcriptionnelle des neuropeptides dans le striatum et donc contribuer à la neuroadaptation du système dopaminergique suivant un traitement aux antipsychotiques typiques. / Neuropeptides (enkephalin and neurotensin) are modulators of dopaminergic system. In the rodent’s striatum, antipsychotic drugs strongly modulate their mRNA levels. For exemple, haloperidol (D2 receptor antagonist) increases their mRNA levels in the striatum, a central structure of dopaminergic system that control locomotor activity. Since haloperidol is associated with many motor side effects, it is likely that neuropeptides modulation is a compensatory mechanism to restore the dopaminergic system homeostasis after D2 receptor blockade. However, molecular mechanism of this transcriptional regulation is not well understood. Nur77 is a transcription factor of the orphan nuclear receptor family that acts as an immediate early gene. In the striatum, its mRNA level is strongly increased following haloperidol treatment. Several evidences suggest that Nur77 is involved in neuropeptides transcriptional modulation. Nur77 can form functional heterodimers with retinoid X receptor (RXR). In agreement with a transcriptional activity of Nur77/RXR complex, RXR agonist (DHA) decreases while the RXR antagonist (HX531) exacerbates acute and chronic motor side effects of haloperidol treatment in wild type mice but remain without effect in Nur77 knockout mice. Our work shows that RXR antagonist (HX531) significantly reduced the increased levels of enkephalin following chronic treatment with haloperidol. We have set up the EMSA (electro-mobility shift assay) to demonstrate the in vitro binding of Nur77 on a responsive element from proenkephalin’s proximal promotor. These results support the hypothesis that Nur77 could be involve in transcriptional modulation of neuropeptides in the striatum and thus, contribute to neuroadaptation of dopaminergic system after treatment with typical antipsychotic drugs.

Submitted by Helmut Patrocinio (hell.kenn@gmail.com) on 2017-11-24T01:32:25Z No. of bitstreams: 1 Cl?udio_Queiroz_Disserta??o.pdf: 793741 bytes, checksum: f79a2a4bb52a8be9ba0b57baec9ed09c (MD5) / Approved for entry into archive by Ismael Pereira (ismael@neuro.ufrn.br) on 2017-11-27T16:10:35Z (GMT) No. of bitstreams: 1 Cl?udio_Queiroz_Disserta??o.pdf: 793741 bytes, checksum: f79a2a4bb52a8be9ba0b57baec9ed09c (MD5) / Made available in DSpace on 2017-11-27T16:11:41Z (GMT). No. of bitstreams: 1 Cl?udio_Queiroz_Disserta??o.pdf: 793741 bytes, checksum: f79a2a4bb52a8be9ba0b57baec9ed09c (MD5) Previous issue date: 1999 / Nos ?ltimos dois s?culos, o conhecimento sobre o sistema nervoso central expandiu-se consideravelmente, possibilitando atualmente o tratamento de muitas patologias do sistema nervoso central. Uma dessas patologias, entretanto, a discinesia tardia n?o apresenta nenhum tratamento terap?utico de efic?cia comprovada (Soares, 1997). A discinesia tardia ? uma s?ndrome caracterizada por movimentos involunt?rios repetitivos, normalmente envolvendo a l?ngua, boca e face, ocasionalmente atingindo tamb?m o pesco?o, membros superiores e quadris. Acredita-se ser a discinesia tardia um efeito colateral da exposi??o prolongada aos antipsic?ticos (neurol?pticos). Essa disfun??o motora pode persistir por meses ou anos ap?s a retirada do tratamento com neurol?ptico, podendo at? mesmo ser irrevers?vel (Karniol, 1979; Casey, 1985; Kane, 1995). Nesta tese de Mestrado, procuramos estudar os efeitos comportamentais da administra??o de buspirona sobre modelos animais de discinesia tardia. Os modelos animais utilizados foram: [1] a supersensibilidade dopamin?rgica induzida por um tratamento prolongado com haloperidol e quantificada pela atividade espont?nea de ratos em um campo aberto e [2] pelo comportamento estereotipado induzido pela apomorfina e [3] a quantifica??o dos movimentos orofaciais de ratos ap?s um tratamento repetido com reserpina. O tratamento prolongado com buspirona per se (3.0 mg/kg, i.p., duas vezes ao dia, por 30 dias) n?o resultou em uma supersensibilidade comportamental em nenhum dos dois modelos animais. O tratamento concomitante de buspirona foi capaz de diminuir os sintomas da supersensibilidade dopamin?rgica induzida pelo haloperidol (2.0 mg/kg, i.p., uma vez ao dia, por 30 dias) e quantificada pela atividade geral em campo aberto, mas n?o pelo comportamento estereotipado induzido pela apomorfina. Nos experimentos agudos, apesar de a buspirona per se diminuir tanto a atividade gera em campo aberto como o comportamento estereotipado induzido pela apomorfina, a co-administra??o de buspirona n?o foi capaz de modificar o efeitos agudos do haloperidol sobre esses dois modelos animais. No terceiro modelo, ratos foram tratados com salina ou buspirona (3.0 mg/kg, i.p., duas vezes ao dia) e ve?culo ou reserpina (0.1 mg/kg, s.c., dias intercalados) por 19 dias. No vig?simo dia, os animais foram observados para a quantifica??o de seus movimentos orofaciais: freq??ncia de protrus?o de l?ngua e movimentos mandibulares e dura??o do tremor facial. O tratamento com buspirona per se n?o foi capaz de induzir a movimentos orofaciais. Animais tratados com reserpina apresentaram maior freq??ncia de movimentos orofaciais em rela??o aos animais tratados com salina. A co-administra??o de buspirona foi capaz de atenuar o desenvolvimento da discinesia orofacial induzida pela reserpina. Verificou-se, tamb?m, que os animais tratados cronicamente com buspirona (3.0 mg/kg, i.p., duas vezes ao dia, 30 dias) desenvolvem maior resposta ao comportamento de bocejo induzido pela apomorfina. Assim, com este trabalho observamos que o tratamento prolongado com buspirona foi capaz de atenuar comportamentos dependentes da disponibilidade de dopamina end?gena (atividade geral em campo aberto e movimentos orofaciais induzidos pela reserpina) provavelmente por meio de uma supersensibilidade dos receptores pr?-sin?pticos (sugerida pelo aumento do comportamento de bocejo induzido por apomorfina). Os dados aqui apresentados, juntamente com a literatura cl?nica existente at? o momento, sugerem um poss?vel papel terap?utico da buspirona no tratamento da discinesia tardia. / In the last two centuries, the knowledgement about the central nervous systems increased enormously, making possible the treatment of patients who suffer of all sort of central nervous systems? diseases. One of this diseases is Tardive Dyskinesia, a syndrome characterized by repetitive involuntary movements, usually involving mouth, face and tongue and sometimes limb and trunk musculature. The syndrome is considered to be an adverse effect of prolonged administration of antipsychotic drugs (normally named neuroleptics). It persists for moths after neuroleptic has been discontinued and may be irreversible (Karniol, 1979; Casey, 1985; Kane, 1995). In a recent meta-analysis study, Soares (1997) concluded that there is no efficacious therapeutic interventions for tardive dyskinesia. In this thesis, we studied the behavior effects of buspirone administration on animal models of tardive dyskinesia. These models comprised the [1] dopaminergic supersensitivity induced by long-term haloperidol administration, which is quantified by the spontaneous activity (locomotion and rearing frequency) of rats observed in an open-field or [2] by the apomorphine-induced stereotyped behavior, and [3] the quantification of orofacial dyskinesia in rats repeatedly treated with reserpine. In the first an second models, buspirone per se (3.0 mg/kg, i.p., twice daily, for 30 days) did not produce dopaminergic supersensitivity. When buspirone was given in combination to haloperidol (2.0 mg/kg, i.p., once daily, for 30 days), it decreased the neuroleptic withdrawal symptoms as detected in open-field but not in apomorphine-induced stereotypy. Although single administration of buspirone per se decreased both open-field and apomorphine-induced stereotypy behavior, buspirone single administration did not modify the acute effects of haloperidol on these two behavioral models. In the third model, rats were co-treated with saline or buspirone (3.0 mg/kg, i.p., twice daily) and vehicle or reserpine (0.1 mg/kg, s.c., once every other day) for 19 days. On the day 20, the animals were observed for the quantification of the behavioral parameters of orofacial dyskinesia: tongue protrusion and vacuous chewing movements frequencies and duration of twitching of the facial musculature. Reserpine-treated rats exhibited a significant increase in the three behavioral parameters of orofacial dyskinesia relative to the saline-treated rats. The co-administration of buspirone in the reserpine-treated rats attenuated the development of orofacial dyskinesia, when compared to the reserpine-treated rats. We also verified that chronic (30 days) buspirone treatment was able to increase apomorphine-induced yawning behavior. The possibility is raised that buspirone attenuates haloperidol-induced increased locomotion and rearing and reserpine-induced orofacial dyskinesia through the development of dopamine autoreceptor supersensitivity. Taken together with previous clinical reports, the present data suggest that buspirone co-administration may lead to important clinical effects concerning different tardive dyskinesia treatment.

Orientadora: Profª Drª Marcela Bermúdez Echeverry / Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Neurociência e Cognição, 2015. / Estudos tem mostrado que o fator de transcricao FosB/ delta FosB e regulado em resposta ao tratamento cronico com antipsicoticos. Porem, so a regulacao do fator FosB no estriado pode ter uma relevancia funcional nas alteracoes motoras pelos antipsicoticos tipicos e atipicos, uma vez que o gene e expressado na via direta e indireta do estriado. Assim, o objetivo do presente trabalho foi determinar se as alteracoes motoras induzidas pela administracao cronica de haloperidol, olanzapina ou clozapina regulam a expressao do FosB de maneira diferenciada no estriado motor e limbico. Metodos: Camundongos adultos machos C57Bl receberam injecao intraperitoneal agudo, subcronico ou cronico de Haloperidol, Olanzapina ou Clozapina. Tambem foram avaliados os efeitos extrapiramidais. Experimento 1: Administracao subcronica (5 dias) foi estudado com haloperidol 4 mg/Kg, olanzapina 15 mg/Kg ou Clozapina 20 mg/Kg para confirmar o efeito cataleptico. Experimento 2: As alteracoes motoras apos do tratamento cronico (77 dias) foram avaliadas com o teste de catalepsia, campo aberto, rota rod, pole test e vacuous chewing movement, com as mesmas doses do experimento 2. No ultimo experimento a expressao da proteina FosB foi avaliada. Resultados: Experimento 1: Com a administracao subcronica tipicos e atipicos induzem a catalepsia, sendo menor com os atipicos. Experimento 2: O tratamento cronico com os dois grupos de neurolepticos induzem alteracoes motoras. Foi observado aumenta da imobilizacao no teste de catalepsia e no campo aberto, sendo maior no grupo Haloperidol. Tambem foi observado alteracao no rota rod. Los grupos Haloperidol e Olanzapina tiveram aumento da expressao do FosB no estriado dorsal e ventral. Porem, o grupo Olanzapina no mostrou aumento na regiao Core do nucleo accumbens, resultado oposto observado com a Clozapina, onde mostrou aumento da expressao na regiao limbica do estriado, sem ser significativo na parte dorsal. Conclusoes: O padrao da expressao do FosB na regiao Core do nucleo accumbens, pode ser correlacionado com os sintomas parkinsonianos induzidos pelos antipsicoticos atipicos como a Clozapina. Assim, a regiao Core, poder ser inclusa na circuitaria dos nucleos basais para o estudo das alteracoes motoras induzidas pelos antipsicoticos. / Background: Studies have shown that the transcription factor FosB/ÄFosB is upregulated in response to chronic neuroleptic treatment. Moreover, regulation of only the factor FosB in striatum might have functional relevance to the motor side effects by either typical or atypical neuroleptics, once this gene is expressed in striatal direct and indirect pathways. Therefore, the goal of this study was determine whether the motor side effects after chronic administration of haloperidol, olanzapine or clozapine shown differential regulation FosB expression in motor and limbic caudate putamen. Methods: Adult male mice C57Bl received either acute, subchronic, or chronic intraperitoneally injections of haloperidol, olanzapine or clozapine, and extrapyramidal effects were evaluated.. Experiment 1: Subchronic administration (5 days) was studied with haloperidol 4 mg/Kg, Olanzapine 15 mg/Kg and clozapine 20 mg/Kg to confirm a steady cataleptic effect. Experiment 2: the motor side effects after chronic treatment (77 days) was evaluated through the catalepsy test, open field, Rota Rod, pole test and vacuous chewing movements, with same doses that experiment 2. Expression FosB protein in the last experiment was examined. Results: Experiment 1: with subchronic administration both typical and atypical neuroleptics induced cataleptic effect, even though the effect was lower with olanzapine and clozapine Experiment 2: chronic treatment showed motor side effects in both neuroleptics groups, evaluated with catalepsy test, open field, with an increased immobilization time in haloperidol group. Also, an alteration in Rota Rod test was observed with both neuroleptics groups, however, haloperidol group was unique showing increased time in pole test, with a little number of vacuous chewing movements but not significant. Haloperidol and olanzapine groups showed increased FosB protein expression in dorsal and ventral striatum. However, olanzapine group had not expression in the Core region in accumbens nucleus, opposite to clozapine group that showed an enlarged expression in this limbic/motor region, with scarcely expression in dorsal striatum. Conclusions: A unique pattern of regulation of FosB expression in core region, accumbens nucleus, may correlates with parkinsonism effect of the atypical neuroleptics, clozapine. Therefore, core region could be included in the basal ganglia circuit to study motor alterations by neuroleptics.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Regular practice of physical activity promotes beneficial effects to the body. However, excessive duration and intensity of exercise may surpass individual tolerance to exercise, generating oxidative stress (OS). Studies have shown these effects in various organs, such as the heart and muscles, but little is known about their action and mechanisms in the brain. Various neurological and neurodegenerative diseases are associated with OS and neurotoxicity. Considering these aspects, the first objective of this study was to determine the influence of chronic moderate exercise in an OS model induced by reserpine in rats. The animals were submitted to daily sessions of swimming, with a gradual increase in the length of training. After eight weeks, the animals received two injections of reserpine or control solutions (1 mg/kg-sc), alternately. A behavioral evaluation was performed, after which the rats were euthanized and the striatum was dissected for enzymatic and biochemical assays. Reserpine increased the vacuous chewing movements frequency (VCM) and facial twitching (FT), as well as catalase activity, but decreased reduced glutathione levels (GSH). Exercise partially prevented FT, and partially recovered GSH levels, but did not modify the effects on catalase and VCM. There was a positive correlation between catalase activity and orofacial dyskinesia (OD) and a negative correlation between GSH and OD. The second objective of this study was to evaluate the effects of an intense physical activity in the same model of OS. Rats were submitted to eleven weeks of swimming (1 h/day), where each rat s load was increased according to its body weight until reaching 7% of its weight. Behavioral evaluations were performed before euthanasia and the striatum was then dissected for assays. The effectiveness of the training was confirmed through reduced levels of serum lactate and cardiac hypertrophy, observed in exercised animals. Intense exercise reduced the locomotor index and exploratory activity of the animals, demonstrating the development of emotional stress. In the presence of reserpine, exercise increased lipid peroxidation (TBARS) and caused an increase in catalase activity, which were positively correlated with each other. Based on the results, it was concluded that chronic physical activity of moderate intensity improved the antioxidant defenses in movement disorders associated with cerebral OS. On the other hand, excessive exercise caused negative emotional disorders and, in the presence of another aggressor agent, modified brain antioxidant capacity, which possibly could aggravate cases of neurological and/or neurodegenerative diseases associated with oxidative processes. / A atividade física praticada de maneira regular promove adaptações benéficas ao organismo, enquanto a inadequação do tempo e intensidade pode exceder a tolerância individual ao exercício gerando estresse oxidativo (EO). Estudos mostram esses efeitos em diversos órgãos como, por exemplo, coração e músculos, mas pouco se conhece sobre sua ação e mecanismos em nível cerebral. Diversas doenças neurológicas e neurodegenerativas estão associadas ao EO e neurotoxicidade. Considerando esses aspectos, o primeiro objetivo desse estudo foi determinar a influência do exercício crônico moderado em modelo de EO induzido por reserpina em ratos. Os animais foram submetidos a sessões diárias de natação com aumento gradual no tempo de treinamento e, após oito semanas, receberam duas doses de solução de reserpina ou controle (1 mg/kg-sc) em dias alternados. Fez-se avaliação comportamental, eutanásia dos animais e retirada da região estriatal do cérebro para determinação enzimática e bioquímica. A reserpina aumentou a freqüência dos movimentos de mascar vazio (MMV) e o tempo de tremor facial (TF); aumentou a atividade da catalase e diminui os níveis de glutationa reduzida (GSH). O exercício preveniu parcialmente o TF e houve recuperação parcial nos níveis de GSH, mas não modificou os efeitos sobre a catalase e MMV. Foi observada uma correlação positiva entre a atividade da catalase e o desenvolvimento de discinesia orofacial (DO), e uma correlação negativa entre GSH e DO. O segundo objetivo desse trabalho foi avaliar os efeitos de uma atividade física intensa sobre este mesmo modelo de EO. Os animais foram submetidos a onze semanas de natação (1 h/dia) com aumento gradual na carga de treinamento até que essa atingisse 7% de seu peso corporal. Realizaram-se as avaliações de comportamento, eutanásia e retirada do estriado para análises. A efetividade do treinamento foi confirmada através dos níveis diminuídos de lactato sérico e do desenvolvimento de hipertrofia cardíaca, observados nos animais exercitados. O exercício intenso reduziu a atividade locomotora e exploratória dos animais, demonstrando desenvolvimento de estresse emocional. Na presença de reserpina, o exercício elevou a peroxidação lipídica (TBARS) e provocou aumento na atividade da catalase, cujos parâmetros apresentaram correlação positiva. Com estes estudos se concluiu que a atividade física crônica de intensidade moderada foi capaz de melhorar as defesas antioxidantes nos distúrbios motores associados ao EO cerebral. Por outro lado, o exercício excessivo provocou alterações emocionais negativas e, quando na presença de um agressor adicional, modificou a capacidade antioxidante do cérebro, o que poderia agravar casos de doenças neurológicas e/ou neurodegenerativas associadas a processos oxidativos.

About 20 to 25 % of psychiatric patients treated with typical antipsychotics may develop an important movement disorder named tardive dyskinesia, with complex etiology and no effective treatment. Cognitive loss is also associated with antipsychotic treatment being harmful and often difficult to identify. Some studies have shown beneficial effects of Omega 3 polyunsaturated fatty acids on tardive dyskinesia and on cognition. Considering the available evidences, we conducted a randomized, double-blind, placebo-controlled, clinical trial, which evaluated the effect of fish oil supplementation rich in n-3 polyunsaturated fatty acids (3 g/dia) or placebo (3 capsules / day), for 12 weeks, on motor and cognitive disturbances of patients on use of neuroleptics. Two evaluations were performed, one before supplementation (baseline) and another at the end of it. For this, were used the following evaluation tools: Abnormal Involuntary Movement Scale for tardive dyskinesia and Mini-Mental State Examination for cognition. Peripheral blood samples were collected before supplementation (baseline) and 4, 8 and 12 weeks after, in order to monitor the supplementation effects on biochemical parameters- triglycerides (TG), fasting glucose, total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and on parameters of blood clotting- prothrombin time and partial thromboplastin time. The biological material collected was also used to run the Comet Assay in leukocytes with the aim of investigate the treatment effect on the DNA damage index. After 3 months of supplementation, we observed that fish oil caused an improvement of 18.5 % in tardive dyskinesia, while cognitive performance did not change when compared with the baseline assessment. Furthermore, lipid profile of the patients was not modified, fasting glucose was reduced in all assessments and prothrombin time increased in the last evaluation. In addition, fish oil did not show genotoxic effect (DNA damage index did not change). Therefore, we have demonstrated the therapeutic potential of fish oil rich in Omega 3 on tardive dyskinesia of patients treated with neuroleptics and its low risk of side effects. The increase of prothrombin time suggests a possible anticoagulant effect, requiring its monitoring during chronic treatment. / Cerca de 20 a 25 % dos pacientes psiquiátricos tratados com antipsicóticos típicos podem desenvolver uma importante síndrome motora denominada discinesia tardia (DT), de etiologia complexa e ainda sem tratamento efetivo. Perdas cognitivas também estão associadas ao uso desses medicamentos, sendo muitas vezes de difícil identificação e igualmente prejudiciais. Alguns estudos têm demonstrado efeitos benéficos dos ácidos graxos poliinsaturados Omega 3 (AGPI n-3) sobre a DT e sobre a cognição. Considerando as evidências disponíveis, realizamos este trabalho que consistiu de um ensaio clínico randomizado, duplo-cego, controlado por placebo. Avaliamos os efeitos da suplementação com de óleo de peixe rico em AGPI n-3 (3 g/dia) ou placebo (3 cápsulas/dia) sobre distúrbios motores e cognitivos de pacientes sob uso de neurolépticos, por um período de 12 semanas. Foram efetuadas duas avaliações, uma antes da suplementação (basal) e outra ao final da mesma. Para isso utilizamos as seguintes ferramentas de avaliação: Escala de Movimentos Involuntários Anormais (EMIA), para a DT e Mini-Exame do Estado Mental (MEEM), para a cognição. Amostras de sangue periférico foram colhidas antes da suplementação (basal) e 4, 8 e 12 semanas após, com o propósito de acompanhar seus os efeitos sobre parâmetros bioquímicos- triglicerídeos (TG), glicemia de jejum, colesterol total, colesterol ligado à lipoproteína de alta densidade (HDL-C) e colesterol ligado à lipoproteína de baixa densidade (LDL-C), bem como sobre parâmetros de coagulação sanguínea- tempo de protrombina (TP) e tempo de tromboplastina parcial (TTP). Com o material biológico colhido, também executamos o Ensaio Cometa em leucócitos, para investigar a influência do tratamento sobre o índice de dano no DNA. Após 3 meses de suplementação, observamos que o óleo de peixe produziu uma melhora de 18,5 % na DT, enquanto o desempenho cognitivo não sofreu alteração em relação à avaliação basal. Por sua vez, o perfil lipídico dos pacientes não foi modificado, a glicemia de jejum foi reduzida em todas as avaliações e o TP aumentou na última avaliação. Ainda, o índice de dano no DNA não foi alterado, mostrando que o óleo de peixe não apresentou efeito genotóxico. Com isso, demonstramos o potencial terapêutico do óleo de peixe rico em Omega 3 sobre a DT de pacientes tratados com neurolépticos e seu baixo risco de efeitos colaterais. O aumento do TP sugere um possível efeito anticoagulante, necessitando de acompanhamento durante tratamentos crônicos.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Chronic treatment of psychotic disorders is associated with adverse effects that affect motor function. Movement disorders include Parkinsonism, akathisia, dystonia and tardive dyskinesia. Magnesium (Mg) is an essential mineral for various physiological functions in the body and its supplementation is used in several diseases. The purpose of this study was investigate the effect of Mg supplementation on the prevention and reversal of orofacial dyskinesia (OD), designated as experiment 1 and 2, respectively, as well as the effect on oxidative stress parameters. In both experiments, male Wistar adult rats were used. In experiment 1, rats were randomly divided into two groups both supplemented with oral solution of magnesium aspartate (40 mg/Kg/mL) or deionized water. After 28 days of supplementation, half of each experimental group was treated with a reserpine solution (0.7 mg/kg/ml, sc) (Mg + R and R groups) or vehicle (C and Mg groups) for 3 days (every other day). One day (24 hours) after the last administration of R/vehicle, all animals were subjected to OD and catalepsy time behavioral assessments. In the experiment 2 rats were randomly divided into two groups and treated with a solution of reserpine (0.7 mg / kg / ml, sc) (R groups) or vehicle (Group C) for 3 days (every other day). Twenty-four hours after the last administration of R/vehicle, the development of OD was quantified. One-half of each experimental group was supplemented immediately once a day (by gavage) with magnesium aspartate (40 mg / kg / ml) (Mg, and groups R + Mg) or deionized water (groups C and R). The OD was measured during subsequent days (every 48 hours). Mg supplementation was maintained throughout the time of behavioral assessment (10 consecutive days). After behavioral evaluations, all animals were euthanized by exsanguination. Blood was drawn for analysis of erythrocytes lipid peroxidation (LP). The brains were immediately dissected for separating cortex, striatum and substantia nigra for determining reactive species (RS) and protein carbonyl (PC). The results showed that Mg supplementation before reserpine administration was sufficient to prevent movement disorder observed in the vacuous chewing movement frequency (VCM) and catalepsy time and also was able to prevent the generation of RS and PC, in both the cortex and the substantia nigra regions, also preventing LP in the erythrocytes. Supplementation of Mg after treatment with reserpine was able to minimize the frequency of VCM and catalepsy time, reduce the generation of RS and PC levels in both cortex and the corpus striatum regions and also reversing the increasing the level of LP in the erythrocytes. Our results underscore the importance of including alternative therapies through supplementation of essential natural substances, like magnesium, which can prevent or ameliorate motor disturbances, often related to chronic treatment of psychotic disorders that so far have no effective treatment. / O tratamento crônico de distúrbios psicóticos está associado a efeitos adversos que afetam a função motora. Os distúrbios do movimento incluem o Parkinsonismo, acatisia, distonias e também discinesia tardia. O magnésio (Mg) é um mineral essencial para diversas funções fisiológicas no organismo e sua suplementação tem sido empregada em diversas doenças. O objetivo deste estudo foi investigar o efeito da suplementação de Mg sobre a prevenção e a reversão da discinesia orofacial (DO), designados como experimento 1 e 2, respectivamente, bem como o efeito sobre parâmetros de estresse oxidativo. Em ambos os experimentos foram utilizados ratos machos Wistar adultos. No experimento 1, os ratos foram divididos aleatoriamente em dois grupos, ambos suplementados oralmente com solução de aspartato de magnésio (40 mg/Kg/mL) ou água deionizada. Depois de 28 dias de suplementação, metade de cada grupo experimental foi tratada com uma solução de reserpina (0,7 mg/kg/mL; sc) (R e Mg + R grupos) ou veículo (C e Mg grupos) durante 3 dias (em dias alternados). Um dia (24 horas) após a última administração de R / veículo, todos os animais foram submetidos a avaliações comportamentais de DO, através da quantificação de movimentos de mascar no vazio (MMV), e tempo de catalepsia. No experimento 2 os ratos foram divididos aleatoriamente em dois grupos e tratados com solução de reserpina (0,7 mg/kg/mL; sc) (grupos R) ou veículo (grupo C) durante 3 dias (em dias alternados). Vinte e quatro horas após a última administração de R / veículo, o desenvolvimento de DO foi quantificada. Metade de cada grupo experimental foi suplementado imediatamente, uma vez por dia (por gavagem) com aspartato de magnésio (40 mg/kg/mL) (grupos Mg e R + Mg) ou água desionizada (grupos C e R). A DO foi quantificada durante os dias subsequentes (cada 48h). A suplementação de Mg foi mantida durante todo o tempo de avaliação comportamental (10 dias consecutivos). Após as avaliações comportamentais todos os animais foram eutanasiados por exsangüinação. O sangue foi retirado para análise dos níveis de lipoperoxidação (LP) eritrocitária. Os cérebros foram imediatamente dissecados para a separação da região do córtex, corpo estriado e substantia nigra para a determinação de espécies reativas (ER) e de proteína carbonil (PC). Os resultados mostraram que a suplementação de Mg antes da administração da reserpina foi suficiente para prevenir os distúrbios do movimento, observados pela frequência de MMV e tempo de catalepsia; bem como foi capaz de evitar a geração de ER e PC, tanto na região do córtex como na substantia nigra, também impedindo a LP nos eritrócitos. A suplementação de Mg após o tratamento com reserpina foi capaz de minimizar a frequência de MMV e o tempo de catalepsia, reduzir a geração de ER e os níveis de PC nas regiões do córtex e do corpo estriado, revertendo também o aumento do nível de LP nos eritrócitos. Nossos resultados ressaltam a importância da inclusão de terapias alternativas através da suplementação de substancias naturais essenciais, como o magnésio, as quais podem prevenir ou atenuar distúrbios motores, frequentemente relacionados ao tratamento crônico de distúrbios psicóticos que até o momento não dispõe de um tratamento eficaz.