Magister Pharmaceuticae - MPharm / Recent scientific findings have highlighted the beneficial roles of polycyclic cage compounds in neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. Further interest into the chemistry of these compounds is stimulated by their remarkable ability to improve the pharmacokinetics profile of known neuroprotective agents. As potent lipophilic scaffolds, they can be employed to target the brain delivery of desired compounds. Inflammation is a key mediator of neuronal cell's degeneration as activated microglia and other inflammatory mediators propagate oxidative damage and neuronal loss. Epidemiological and clinical evidence suggests that non-steroidal anti-inflammatory drugs (NSAIDs) slow down the progression and onset of neurodegenerative diseases. The beneficial effects of NSAIDs in ND can be attributed to their ability to inhibit cyclooxygenase enzymes thereby halting the biosynthesis of prostaglandins (PG) which are powerful mediators of inflammation. NSAIDs also inhibit the expression of pro- inflammatory genes. Despite their potential neuroprotective activity, NSAIDs are poorly lipophilic due to the presence of polar carboxylic acid groups and will therefore ionise at physiological pH, deterring them from reaching the desired site of action in the central nervous system (CNS).

A Doença de Alzheimer (DA) é uma doença cerebral progressiva que resulta em prejuízos na memória e disfunção cognitiva global. Entre as principais características neuropatológicas associadas a DA estão à presença de placas senis, emaranhados neurofibrilares e a hiperfosforilação da proteína Tau. A hiperativação do sistema glutamatérgico tem sido implicada na fisiopatologia da DA. O excesso de glutamato na fenda sináptica causa hiperativação do seu receptor ionótropico N-metill-D-aspartato (NMDA) o que favorece o aumento do influxo de cálcio e morte neuronal. A administração intracerebral de ácido ocadáico (AO) causa alterações morfológicas e funcionais similares à DA. O AO promove a inibição da proteína fosfatase 2A (PP2A) favorecendo as atividades cinásicas de proteínas como a cinase dependente de ciclina 5 (Cdk5). A memantina (MN) é uma das principais drogas utilizadas no tratamento da DA e o seu mecanismo de ação envolve um antagonismo não competitivo de baixa afinidade pela subunidade NR2B do receptor NMDA. Neste trabalho, foram avaliados efeitos do pré-tratamento com MN em um modelo semelhante a DA induzido pela administração intrahipocampal de AO em ratos. O pré-tratamento com MN preveniu o déficit na memória especial causado pela infusão intrahipocampal de AO. Os mecanismos envolvidos nestes efeitos neuroprotetores envolvem a prevenção do aumento de glutamato no liquido cefalorraquidiano, juntamente com a regulação da expressão de Cdk5 e em conseqüência a prevenção do aumento da fosforilação de Tau. Desta maneira, a MN pode ser um alvo terapêutico para prevenir as alterações comportamentais e neuroquímicas em um modelo similar a DA induzido pelo AO. / Alzheimer's disease (AD) is a progressive brain disease that causes memory loss and global cognitive dysfunction. The neuropathological alterations associated with AD include senile plaques, neurofibrillary tangles and Tau protein hyperphosphorylation. The glutamatergic system is implicated in the pathophysiology of AD. Indeed, the excessive glutamate levels in the synaptic cleft may cause hyperactivation of glutamate ionotropic N-metill-Daspartate (NMDA), which favors increase calcium influx and neuronal death. The intracerebral administration of okadaic acid (OA) causes morphological and functional alterations similar to AD. The OA inhibits the protein phosphatase 2A (PP2A) thus overstimulating the kinases activities. Memantine (MN) is a drug currently used in the treatment of AD, which mechanism involves a noncompetitive low affinity antagonism for NR2B subunit of NMDA receptors. In this work we evaluate the effects of pretreatment with MN in an AD-like model in rats induced by intrahippocampal administration of OA. The pretreatment with MN could prevent the spatial memory deficits caused by OA intrahipocampal administration in rats. The mechanisms underlying this neuroprotective effects involves the prevention of the increase in brain glutamate levels along with regulation of Cdk5 and, in consequence, downstream phosphorylation of Tau (ser199/202) protein. To conclude, MN has potential therapeutic role in preventing behavioral and neurochemical alterations caused by an AD like model induced by OA.

Prion diseases are fatal transmisible neurodegenerative and infectious disorders (TSEs) of humans and animals, characterized by structural transition of the host-encoded cellular prion protein (PrPC) into the aberrantly folded pathologic isoform PrPSc. The main aim of this work is to summarize present information about prion diseases and their possibilities of determination pointed to electrochemical techniques. For this purpose cyclic voltammetry (CV), differential pulse voltammetry, differential pulse voltammetry Brdicka reaction and chronopotentiometric stripping analysis (CPSA) were used. The estimated detection limits were 32 ug/ml by CV, 16 ug/ml by DPV, 16 ug/ml by DPV -- Brdicka reaction and 8 ug/ml by CPSA. Subsequently, the influence of heat denaturation was observed. It clearly follows from the obtained results that signals of prion decreased linearly depending on the duration of the heat treatment at 99°C for various time intervals 0, 15, 30, 45, and 60 min. Moreover, we aimed our attention on studying of prion protein interaction with CdTe quantum dots (QDs) using electrochemistry. Primarily, we characterized electrochemical properties of QDs and the detection limit at 100 fg/ml was estimated. Further, electrochemical study of prion and QD interactions was carried out to find the most suitable conditions for sensitive detection of prion proteins. Detection limit (3 S/N) was estimated as 1 fg in 5 ul. This makes labeling of proteins with QDs of great importance due to easy applicability and possibility to use in miniaturized devices, which can be used in situ. This should open new possibilities how to determine the presence of these proteins on surgical equipment and other types of materials, which could be contagious.

Le concept de "cognition incarnée" considère que le schéma classique Perception-Cognition-Action proposant un flux séquentiel de traitement de l’information n'est pas approprié pour comprendre l'effet comportemental des troubles neurodégénératifs et trouver des solutions thérapeutiques innovantes. La découverte des neurones miroirs (NM) a donné un substrat biologique à cette théorie: on pense maintenant que les NM relient les connaissances sur les actions et les perceptions non seulement pour intégrer la perception dans la planification et l'exécution, mais aussi pour soutenir un large éventail de fonctions cognitives, par ex. empathie et langage. En même temps, il est maintenant clair que dans chaque maladie neurodégénérative les symptômes cognitifs et moteurs sont représentés le long d'un continuum. Les maladies neurodégénératives liées au vieillissement, comme la maladie d'Alzheimer (MA), la forme la plus courante de démence, sont devenues un enjeu social très important. Comme il n'y a pas de remède pour la MA, les études se concentrent sur la prévention. Une catégorie qui représente maintenant une cible privilégiée est le trouble cognitif léger (TCL), considéré comme une étape intermédiaire entre le vieillissement normal et la MA. Même si MA et TCL ont été caractérisées comme des maladies «cognitives» jusqu'à présent, un lien entre la fonction motrice et le risque de développer la MA a été reconnu.Le but principal de cette recherche est d'étudier l'intégrité du réseau NM dans la MA, le TCL et le vieillissement normal. La caractérisation de son fonctionnement dans les maladies neurodégénératives serait utile pour une meilleure compréhension de leurs mécanismes fonctionnels et manifestations cliniques. Cela permettrait également d’exploiter le NM dans la réhabilitation des symptômes.La thèse comprend deux parties : la première inclue une vaste recherche bibliographique destinée à décrire le cadre scientifique qui justifie une telle recherche.Nous avons d'abord passé en revue les preuves sur l'existence d'un système NM chez les singes et les humains, et ses multiples rôles possibles et après brièvement décrit le tableau clinique des principaux troubles neurodégénératifs, en montrant comment les symptômes cognitifs et moteurs s’entrecroisent. Ensuite, nous avons détaillé les résultats de la recherche documentaire sur les maladies neurodégénératives, NM et cognition incarnée, en les commentant à la lumière de cette théorie.La deuxième partie de la thèse décrit la procédure expérimentale qui a été réalisée dans le but de la recherche.Trois groupes appariés de 16 sujets chacun (CA-contrôles âgées, TCL amnésique avec atrophie hippocampique et MA) ont été évalués avec une batterie neuropsychologique centrée sur les fonctions liées au système NM, et une tâche IRMf spécifiquement créée pour tester les NM: celle- ci était constituée d'une tache d’observation, où aux sujets ont été montrés des vidéos d'une main droite saisissant différents objets, et d'une tache motrice où les sujets ont observé des images d'objets orientés pour être saisis avec la main droite, et ont fait le geste correspondant.Chez les CA, l'analyse de conjonction (comparant l'activation de l'IRMf pendant l'observation et l'exécution) a indiqué l'activation d'un réseau bilatéral fronto-pariétal dans les zones NM « classiques» et du gyrus temporal supérieur (STG), entrée visuelle corticale aux NM. Le groupe TCL a montré une activation similaire, cependant, les zones pariétales ont été moins activées et le STG n'a pas été activé, tandis que l'inverse était vrai pour la zone de Broca droite. Nous n'avons observé aucune activation du réseau fronto-pariétal chez le groupe MA. Dans tous les tests neuropsychologiques (y compris les tests de fonctions attribuées à NM), les sujets MA ont été plus mauvais que les CA, alors que les sujets TCL montraient seulement des troubles de mémoire épisodique et fluidité sémantique (...). / The concept of “embodied cognition” considers that the classical Perception-Cognition-Action architecture proposing a sequential flow of processing with clean cuts between all modules is not appropriate to understand the behavioral effect of neurodegenerative disorders and to find innovative therapeutic solutions. In the last decades, the discovery of the mirror neurons (MN) has given a biological substrate to this theoretical perspective: the MN are now thought linking together knowledge about actions and perceptions not only to integrate perception in action planning and execution but also as a neural mechanism supporting a wide range of cognitive functions, e.g. empathy and language. At the same time, it is now clear that in each neurodegenerative disease both cognitive and motor symptoms are represented along a continuum. In the current demographic context, neurodegenerative diseases linked to aging have become a very important social issue. Alzheimer Disease (AD), the most common form of dementia, is a neurodegenerative disease strictly linked to aging. As actually there is no cure, several studies are focusing on prevention. A category which now represents a preferential target of intervention is Mild Cognitive Impairment (MCI), considered as an intermediate stage between normal aging and AD. Even if AD and MCI have been characterized as “cognitive” diseases until now, a link between motor function and the risk of developing AD has been recognized.The main purpose of this research is to investigate the integrity of the MN network in AD, MCI and normal aging. Characterizing the functioning of the MN network in neurodegenerative diseases would be useful to better understand functional mechanisms and their clinical manifestations. It would also allow to capitalize on these kinds of neurons in the rehabilitation of motor and cognitive symptoms.The thesis consists of two parts: the first part includes an extensive bibliographic research intended to describe the scientific frame which justifies such a research.We first reviewed the evidence about the existence of a MN system in monkeys and humans, and its multiple possible roles in humans.We then briefly reviewed the clinical picture of the main neurodegenerative disorders, showing how cognitive and motor symptoms intersect in all of them.Next, we detailed the results of literature searching on neurodegenerative diseases, MN, and embodied cognition, commenting them at the light of this hypothesis.The second part of the thesis describe the experimental procedure which has been performed to evaluate the integrity of the MN network in normal elderly and people with AD and MCI, and its results.Three matched groups of 16 subjects each (normal elderly-NE, amnesic MCI with hippocampal atrophy and AD) were evaluated with a neuropsychological battery centered on functions thought to be linked to the MN system, and a fMRI task specifically created to test MN: that comprised of an observation run, where subjects were shown videos of a right hand grasping different objects, and of a motor run, where subjects observed visual pictures of objects oriented to be grasped with the right hand, and made the corresponding gesture.In NE subjects, the conjunction analysis (comparing fMRI activation during observation and execution), indicated the activation of a bilateral fronto-parietal network in “classical” MN areas, and of the superior temporal gyrus (STG), an area thought to provide the cortical visual input to the MN. The MCI group showed the activation of areas belonging to the same network, however, parietal areas were activated to a lesser extent and the STG was not activated, while the opposite was true for the right Broca’s area. We did not observe any activation of the fronto-parietal network in AD participants (...).

abstract: Several debilitating neurological disorders, such as Alzheimer's disease, stroke, and spinal cord injury, are characterized by the damage or loss of neuronal cell types in the central nervous system (CNS). Human neural progenitor cells (hNPCs) derived from human pluripotent stem cells (hPSCs) can proliferate extensively and differentiate into the various neuronal subtypes and supporting cells that comprise the CNS. As such, hNPCs have tremendous potential for disease modeling, drug screening, and regenerative medicine applications. However, the use hNPCs for the study and treatment of neurological diseases requires the development of defined, robust, and scalable methods for their expansion and neuronal differentiation. To that end a rational design process was used to develop a vitronectin-derived peptide (VDP)-based substrate to support the growth and neuronal differentiation of hNPCs in conventional two-dimensional (2-D) culture and large-scale microcarrier (MC)-based suspension culture. Compared to hNPCs cultured on ECMP-based substrates, hNPCs grown on VDP-coated surfaces displayed similar morphologies, growth rates, and high expression levels of hNPC multipotency markers. Furthermore, VDP surfaces supported the directed differentiation of hNPCs to neurons at similar levels to cells differentiated on ECMP substrates. Here it has been demonstrated that VDP is a robust growth and differentiation matrix, as demonstrated by its ability to support the expansions and neuronal differentiation of hNPCs derived from three hESC (H9, HUES9, and HSF4) and one hiPSC (RiPSC) cell lines. Finally, it has been shown that VDP allows for the expansion or neuronal differentiation of hNPCs to quantities (>1010) necessary for drug screening or regenerative medicine purposes. In the future, the use of VDP as a defined culture substrate will significantly advance the clinical application of hNPCs and their derivatives as it will enable the large-scale expansion and neuronal differentiation of hNPCs in quantities necessary for disease modeling, drug screening, and regenerative medicine applications. / Dissertation/Thesis / Masters Thesis Bioengineering 2016

A Doença de Alzheimer (DA) é uma doença cerebral progressiva que resulta em prejuízos na memória e disfunção cognitiva global. Entre as principais características neuropatológicas associadas a DA estão à presença de placas senis, emaranhados neurofibrilares e a hiperfosforilação da proteína Tau. A hiperativação do sistema glutamatérgico tem sido implicada na fisiopatologia da DA. O excesso de glutamato na fenda sináptica causa hiperativação do seu receptor ionótropico N-metill-D-aspartato (NMDA) o que favorece o aumento do influxo de cálcio e morte neuronal. A administração intracerebral de ácido ocadáico (AO) causa alterações morfológicas e funcionais similares à DA. O AO promove a inibição da proteína fosfatase 2A (PP2A) favorecendo as atividades cinásicas de proteínas como a cinase dependente de ciclina 5 (Cdk5). A memantina (MN) é uma das principais drogas utilizadas no tratamento da DA e o seu mecanismo de ação envolve um antagonismo não competitivo de baixa afinidade pela subunidade NR2B do receptor NMDA. Neste trabalho, foram avaliados efeitos do pré-tratamento com MN em um modelo semelhante a DA induzido pela administração intrahipocampal de AO em ratos. O pré-tratamento com MN preveniu o déficit na memória especial causado pela infusão intrahipocampal de AO. Os mecanismos envolvidos nestes efeitos neuroprotetores envolvem a prevenção do aumento de glutamato no liquido cefalorraquidiano, juntamente com a regulação da expressão de Cdk5 e em conseqüência a prevenção do aumento da fosforilação de Tau. Desta maneira, a MN pode ser um alvo terapêutico para prevenir as alterações comportamentais e neuroquímicas em um modelo similar a DA induzido pelo AO. / Alzheimer's disease (AD) is a progressive brain disease that causes memory loss and global cognitive dysfunction. The neuropathological alterations associated with AD include senile plaques, neurofibrillary tangles and Tau protein hyperphosphorylation. The glutamatergic system is implicated in the pathophysiology of AD. Indeed, the excessive glutamate levels in the synaptic cleft may cause hyperactivation of glutamate ionotropic N-metill-Daspartate (NMDA), which favors increase calcium influx and neuronal death. The intracerebral administration of okadaic acid (OA) causes morphological and functional alterations similar to AD. The OA inhibits the protein phosphatase 2A (PP2A) thus overstimulating the kinases activities. Memantine (MN) is a drug currently used in the treatment of AD, which mechanism involves a noncompetitive low affinity antagonism for NR2B subunit of NMDA receptors. In this work we evaluate the effects of pretreatment with MN in an AD-like model in rats induced by intrahippocampal administration of OA. The pretreatment with MN could prevent the spatial memory deficits caused by OA intrahipocampal administration in rats. The mechanisms underlying this neuroprotective effects involves the prevention of the increase in brain glutamate levels along with regulation of Cdk5 and, in consequence, downstream phosphorylation of Tau (ser199/202) protein. To conclude, MN has potential therapeutic role in preventing behavioral and neurochemical alterations caused by an AD like model induced by OA.

Les maladies à Prions sont des maladies neurodégénératives fatales, touchant l'homme et l'animal. Même si le risque de transmission de la maladie de la vache folle à l'homme semble maîtrisé, il persiste actuellement un risque de santé publique lié à la transmission iatrogène de cette forme, notamment par transfusion sanguine. Pour contrôler cette transmission, il est donc essentiel de mieux comprendre les mécanismes moléculaires et cellulaires de réplication et de dissémination des Prions. Ces mécanismes de réplication se produisent à des échelles de temps et de taille difficilement accessibles expérimentalement, et ont ainsi fait l'objet de nombreuses modélisations théoriques utiles pour aider à la compréhension des mécanismes. L'objectif de cette thèse est de compléter ces modèles mathématiques, afin d'étudier plus spécifiquement les conséquences dynamiques sur la réplication des Prions, des propriétés de réplication taille-dépendante d'une part, et de l'hétérogénéité des cellules impliquées dans la réplication d'autre part. Dans un premier temps, nous avons généralisé un modèle de polymérisation nucléée pour prendre en compte un taux d'élongation des fibrilles dépendant de leur taille. Nous avons principalement déduit de cette étude que la distribution en taille des agrégats semble une donnée expérimentale très informative sur les mécanismes élémentaires de réplication, au contraire du profil cinétique d'accumulation de la PrPres peu sensibles aux propriétés de réplication taille-dépendantes. Dans un second temps, après une caractérisation expérimentale de l'hétérogénéité cellulaire de réplication, nous avons intégré le mécanisme de réplication intracellulaire à un modèle multicellulaire par automate cellulaire continu stochastique. De manière appliquée, cette étude nous a permis d'identifier des étapes du processus de culture cellulaire critiques pour l'établissement d'une infection chronique, et nous a permis de proposer plusieurs protocoles pour augmenter la sensibilité des cultures cellulaires aux infections à Prions. / Prion diseases are neurodegenerative, fatal and transmissible diseases, with no effective treatment. The risk of transmission of bovine spongiform encephalopathy to humans is now under control ; however the risk of human-to-human transmission of variant Creutzfeldt-Jakob disease via medical treatments (notably through blood transfusion) remains. Thus, understanding cellular and molecular mechanisms responsible for Prion replication and dissemination is critical to efficiently control Prion transmission. The mechanisms of Prion replication are poorly characterised and occur at time and size scale achieved experimentally with difficulty. Thus, mathematical models can help us understand prion multiplication by testing which mechanisms best fit to experimental data. Therefore the objectives of our study are to complete existing mathematical model in order to investigate the size-dependent replicative properties of prion aggregates and the cellular heterogeneity. Firstly, we have extended a previous study of the nucleated polymerization model to take into account size dependent replicative properties of prion aggregates. This is achieved by a choice of coefficients in the model that are not constant. Our results suggest that the size distribution of prion aggregates could be one of the most informative experimental data to study elementary replication mechanisms and to investigate strain phenomenon. Secondly, we have modelled the multicellular dynamics of prion replication by integrating intracellular replication (by nucleated polymerization) into a continuous and stochastic cellular automaton. The model formulation is based on an experimental characterisation of cellular heterogeneity. From an applicative point of view, this theoretical study has allowed us to propose several protocols to increase cell culture sensitivity to prion infection.

No presente trabalho foram avaliados os efeitos da estimulação em ambiente enriquecido (AE) para a memória e para os marcadores neuropatológicos da Doença de Alzheimer (DA) em camundongos transgênicos (TG) que superexpressam a proteína precursora de amilóide humana. Nos animais TG ou C57Bl/6, estimulados dos 3 aos 7 meses de idade, o AE promoveu manutenção da memória relacionada a estímulo aversivo por até 2 meses. Em animais mais velhos (8 a 12 meses de idade), a manutenção da memória foi verificada, independente do estímulo. Em ambas as idades, a memória espacial não foi alterada com o AE. Não houve alteração do número de placas senis nos animais mais jovens. Porém, em TG de 12 meses estimulados, houve redução de 85% na densidade de placas no hipocampo quando comparados ao grupo TG controle. Nessa idade, porém, não foi possível identificar a formação de enovelamentos neurofibrilares. Em conclusão, a estimulação crônica em ambiente enriquecido contribuiu para a formação de reservas cognitivas, protetoras no caso de doenças neurodegenerativas como a DA. / In the present work the effects of chronic stimulation in enriched envitonment (EE) for memory and neuropathological hallmarks of the Alzheimer\'s disease (AD) were evaluated in transgenic mice (TG) that overexpress the human amyloid precursor protein. In TG or C57Bl/6 animals, stimulated from 3 to 7 months of age, submition in EE kept fear conditioning memory for as long as two months. In older mice (8 to 12 months of age) memory keeping was verified, independent of the stimulus. In both ages, spatial memory was not altered with EE. There was no difference in senile plaques density in younger animals. However, in stimulated 12-months-old animals, there was a reduction of 85% in the density of senile plaques in hippocampus, when compared to TG control animals. In this age, however, it was not possible to identify neurofibrilary tangles. In conclusion, chronic stimulation in enriched environment contributed to formation of cognitive reserves, protective in the presence of neurodegenerative diseases, as AD.

A Doença de Alzheimer (DA) é uma doença cerebral progressiva que resulta em prejuízos na memória e disfunção cognitiva global. Entre as principais características neuropatológicas associadas a DA estão à presença de placas senis, emaranhados neurofibrilares e a hiperfosforilação da proteína Tau. A hiperativação do sistema glutamatérgico tem sido implicada na fisiopatologia da DA. O excesso de glutamato na fenda sináptica causa hiperativação do seu receptor ionótropico N-metill-D-aspartato (NMDA) o que favorece o aumento do influxo de cálcio e morte neuronal. A administração intracerebral de ácido ocadáico (AO) causa alterações morfológicas e funcionais similares à DA. O AO promove a inibição da proteína fosfatase 2A (PP2A) favorecendo as atividades cinásicas de proteínas como a cinase dependente de ciclina 5 (Cdk5). A memantina (MN) é uma das principais drogas utilizadas no tratamento da DA e o seu mecanismo de ação envolve um antagonismo não competitivo de baixa afinidade pela subunidade NR2B do receptor NMDA. Neste trabalho, foram avaliados efeitos do pré-tratamento com MN em um modelo semelhante a DA induzido pela administração intrahipocampal de AO em ratos. O pré-tratamento com MN preveniu o déficit na memória especial causado pela infusão intrahipocampal de AO. Os mecanismos envolvidos nestes efeitos neuroprotetores envolvem a prevenção do aumento de glutamato no liquido cefalorraquidiano, juntamente com a regulação da expressão de Cdk5 e em conseqüência a prevenção do aumento da fosforilação de Tau. Desta maneira, a MN pode ser um alvo terapêutico para prevenir as alterações comportamentais e neuroquímicas em um modelo similar a DA induzido pelo AO. / Alzheimer's disease (AD) is a progressive brain disease that causes memory loss and global cognitive dysfunction. The neuropathological alterations associated with AD include senile plaques, neurofibrillary tangles and Tau protein hyperphosphorylation. The glutamatergic system is implicated in the pathophysiology of AD. Indeed, the excessive glutamate levels in the synaptic cleft may cause hyperactivation of glutamate ionotropic N-metill-Daspartate (NMDA), which favors increase calcium influx and neuronal death. The intracerebral administration of okadaic acid (OA) causes morphological and functional alterations similar to AD. The OA inhibits the protein phosphatase 2A (PP2A) thus overstimulating the kinases activities. Memantine (MN) is a drug currently used in the treatment of AD, which mechanism involves a noncompetitive low affinity antagonism for NR2B subunit of NMDA receptors. In this work we evaluate the effects of pretreatment with MN in an AD-like model in rats induced by intrahippocampal administration of OA. The pretreatment with MN could prevent the spatial memory deficits caused by OA intrahipocampal administration in rats. The mechanisms underlying this neuroprotective effects involves the prevention of the increase in brain glutamate levels along with regulation of Cdk5 and, in consequence, downstream phosphorylation of Tau (ser199/202) protein. To conclude, MN has potential therapeutic role in preventing behavioral and neurochemical alterations caused by an AD like model induced by OA.

As doenças neurodegenerativas (DN) estão entre as principais causas de mortalidade e morbidade nos países ocidentais. Não há ainda um tratamento definitivo para estas neuropatias, mas os estudos têm indicado mecanismos comuns de toxicidade que incluem disfunção mitocondrial, estresse oxidativo e apoptose. Assim, as mitocôndrias constituem alvos importantes para futuras estratégias de neuroproteção a fim de tratar, prevenir ou até mesmo retardar a neurodegeneração. Neste contexto, o canabidiol (CBD), um constituinte não psicoativo da Cannabis sativa e cuja propriedade neuroprotetora tem sido sugerida por diferentes estudos, surge como uma alternativa bastante promissora. Diferentes mecanismos moleculares podem estar envolvidos na neuroproteção exercida pelo CBD. Embora o potencial efeito benéfico do canabidiol com relação às doenças neurodegenerativas já tenha sido sugerido, não há ainda estudos que abordem precisamente os mecanismos de proteção contra a toxicidade mitocondrial cerebral, evento chave no processo neurodegenerativo. O presente estudo teve como objetivo investigar os efeitos do CBD em mitocôndrias cerebrais de rato, bem como possíveis mecanismos de neuroproteção. Foram avaliados os seguintes parâmetros: função mitocondrial, estresse oxidativo mitocondrial e transição de permeabilidade de membrana mitocondrial (TPMM). Os resultados obtidos sugerem que o canabidiol é capaz de proteger as mitocôndrias cerebrais contra o intumescimento osmótico induzido por cálcio/fosfato, contra a produção de H2O2 induzida por terc-butil hidroperóxido e contra a peroxidação lipídica induzida por Fe2+ e citrato. A captação mitocondrial de cálcio e a capacidade fosforilativa não foram afetadas. / Neurodegenerative diseases (ND) are among the leading causes of mortality and morbidity in Western countries. There is not a definitive treatment for these neuropathies, but studies have indicated mechanisms of toxicity which include mitochondrial dysfunction, oxidative stress and apoptosis. Therefore, mitochondria are important targets for future neuroprotective strategies to treat, prevent or even slow the neurodegeneration. In this context, cannabidiol (CBD), a constituent of non-psychoactive Cannabis sativa and whose neuroprotective property has been suggested by different studies, emerges as a promising alternative. Different molecular mechanisms may be involved in the neuroprotection exerted by CBD. Although the potential beneficial effects of cannabidiol in relation to neurodegenerative diseases has already been suggested, there are no studies addressing specifically the mechanisms of protection against mitochondrial toxicity brain, a key event in the neurodegenerative process. This study aimed to investigate the effects of CBD on rat brain mitochondria, as well as the mechanisms of neuroprotection. The following parameters were evaluated: mitochondrial function, mitochondrial oxidative stress and permeability transition of the mitochondrial membrane (MPT). The results suggest that cannabidiol can protect brain mitochondria against: the osmotic swelling induced by calcium/phosphate, the production of H2O2 induced by tert-butyl hydroperoxide and the lipid peroxidation induced by Fe2+ and citrate. The mitochondrial calcium uptake and phosphorylative capacity were not affected.