The neuroendocrine pathways and mechanisms for the control of the reproduction in female pigs

Abstract Within the hypothalamic-pituitary-gonad (HPG) axis, the major hierarchical component is gonadotropin-releasing hormone (GnRH) neurons, which directly or indirectly receive regulatory inputs from a wide array of regulatory signals and pathways, involving numerous circulating hormones, neuropeptides, and neurotransmitters, and which operate as a final output for the brain control of reproduction. In recent years, there has been an increasing interest in neuropeptides that have the potential to stimulate or inhibit GnRH in the hypothalamus of pigs. Among them, Kisspeptin is a key component in the precise regulation of GnRH neuron secretion activity. Besides, other neuropeptides, including neurokinin B (NKB), neuromedin B (NMB), neuromedin S (NMS), α-melanocyte-stimulating hormone (α-MSH), Phoenixin (PNX), show potential for having a stimulating effect on GnRH neurons. On the contrary, RFamide-related peptide-3 (RFRP-3), endogenous opioid peptides (EOP), neuropeptide Y (NPY), and Galanin (GAL) may play an inhibitory role in the regulation of porcine reproductive nerves and may directly or indirectly regulate GnRH neurons. By combining data from suitable model species and pigs, we aim to provide a comprehensive summary of our current understanding of the neuropeptides acting on GnRH neurons, with a particular focus on their central regulatory pathways and underlying molecular basis.

The neuroendocrine pathways and mechanisms for the control of the reproduction in female pigs

Within the hypothalamic-pituitary-gonad (HPG) axis, the major hierarchical component is gonadotropin-releasing hormone (GnRH) neurons, which directly or indirectly receive regulatory inputs from a wide array of regulatory signals and pathways, involving numerous circulating hormones, neuropeptides, and neurotransmitters, and which operate as a final output for the brain control of reproduction. In recent years, there has been an increasing interest in neuropeptides that have the potential to stimulate or inhibit GnRH in the hypothalamus of pigs. Among them, Kisspeptin is a key component in the precise regulation of GnRH neuron secretion activity. Besides, other neuropeptides, including neurokinin B (NKB), neuromedin B (NMB), neuromedin S (NMS), α-melanocyte-stimulating hormone (α-MSH), Phoenixin (PNX), show potential for having a stimulating effect on GnRH neurons. On the contrary, RFamide-related peptide-3 (RFRP-3), endogenous opioid peptides (EOP), neuropeptide Y (NPY), and Galanin (GAL) may play an inhibitory role in the regulation of porcine reproductive nerves and may directly or indirectly regulate GnRH neurons. By combining data from suitable model species and pigs, we aim to provide a comprehensive summary of our current understanding of the neuropeptides acting on GnRH neurons, with a particular focus on their central regulatory pathways and underlying molecular basis.(AU)

Síndrome do mau ajustamento neonatal em potros: foco em neuroesteróides / Foal mal adjustment syndrome: focus on neurosteroids

O processo de transição do feto para a vida extra-uterina é considerado um período crítico que requer complexas adaptações fisiológicas do potro neonato. Eventos estressores de origem hipóxicoisquêmicas no periparto podem desencadear um quadro de encefalopatia neonatal equina, também conhecida como síndrome do mau ajustamento neonatal. O diagnóstico é feito baseado na avaliação clínica e na anamnese e avaliação do histórico da gestação. Casos leves a moderados tem prognóstico favorável. É imprescindível o entendimento da endocrinologia da gestação, do papel dos neuroesteróides no desenvolvimento do sistema nervoso fetal para que o estabelecimento precoce da terapia adequada seja realizado de maneira bem sucedida. Assim, o objetivo do presente é abordar os principais aspectos clínicos e fisiopatológicos da Síndrome do Mau Ajustamento Neonatal em neonatos equinos, com foco especial no papel dos neuroesteróides durante a maturação cerebral do feto no terço final da gestação e na transição para a vida extra-uterina.

The transition from fetus to extrauterine life is considered a critical period that requires complex physiological adaptations on the part of the newborn foal. Peripartum hypoxic-ischemic stressors can result in equine neonatal encephalopathy, also known to as neonatal maladjustment syndrome. The diagnosis is made based on clinical examination, anamnesis, and a review of the mare’s pregnancy history. Cases that are mild to moderate in severity have a favorable prognosis. It is critical to understand the endocrinology of pregnancy and the role of neurosteroids in the development of the fetal nervous system in order to successfully initiate appropriate therapy early. Thus, the purpose of this article is to discuss the major clinical and pathophysiological aspects of neonatal maladjustment syndrome in equine neonates, with a particular emphasis on the role of neurosteroids during fetal brain maturation in the final third of pregnancy and during the transition to extrauterine life.

Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation

Tetanus toxin blocks the release of the inhibitory neurotransmitters in the central nervous system and causes tetanus and its main form of prevention is through vaccination. The vaccine is produced by inactivation of tetanus toxin with formaldehyde, which may cause side effects. An alternative way is the use of ionizing radiation for inactivation of the toxin and also to improve the potential immunogenic response and to reduce the post-vaccination side effects. Therefore, the aim of this study was to characterize the tetanus toxin structure after different doses of ionizing radiation of 60Co. Methods Irradiated and native tetanus toxin was characterized by SDS PAGE in reducing and non-reducing conditions and MALD-TOF. Enzymatic activity was measured by FRET substrate. Also, antigenic properties were assessed by ELISA and Western Blot data. Results Characterization analysis revealed gradual modification on the tetanus toxin structure according to doses increase. Also, fragmentation and possible aggregations of the protein fragments were observed in higher doses. In the analysis of peptide preservation by enzymatic digestion and mass spectrometry, there was a slight modification in the identification up to the dose of 4 kGy. At subsequent doses, peptide identification was minimal. The analysis of the enzymatic activity by fluorescence showed 35 % attenuation in the activity even at higher doses. In the antigenic evaluation, anti-tetanus toxin antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels. Conclusion Ionizing radiation promoted structural changes in the tetanus toxin such as fragmentation and/or aggregation and attenuation of enzymatic activity as the dose increased, but antigenic recognition of the toxin remained at good levels indicating its possible use as an immunogen. However, studies of enzymatic activity of tetanus toxin irradiated with doses above 8 kGy should be further analyzed.(AU)

Characterization and evaluation of the enzymatic activity of tetanus toxin submitted to cobalt-60 gamma radiation

Tetanus toxin blocks the release of the inhibitory neurotransmitters in the central nervous system and causes tetanus and its main form of prevention is through vaccination. The vaccine is produced by inactivation of tetanus toxin with formaldehyde, which may cause side effects. An alternative way is the use of ionizing radiation for inactivation of the toxin and also to improve the potential immunogenic response and to reduce the post-vaccination side effects. Therefore, the aim of this study was to characterize the tetanus toxin structure after different doses of ionizing radiation of 60Co. Methods Irradiated and native tetanus toxin was characterized by SDS PAGE in reducing and non-reducing conditions and MALD-TOF. Enzymatic activity was measured by FRET substrate. Also, antigenic properties were assessed by ELISA and Western Blot data. Results Characterization analysis revealed gradual modification on the tetanus toxin structure according to doses increase. Also, fragmentation and possible aggregations of the protein fragments were observed in higher doses. In the analysis of peptide preservation by enzymatic digestion and mass spectrometry, there was a slight modification in the identification up to the dose of 4 kGy. At subsequent doses, peptide identification was minimal. The analysis of the enzymatic activity by fluorescence showed 35 % attenuation in the activity even at higher doses. In the antigenic evaluation, anti-tetanus toxin antibodies were detected against the irradiated toxins at the different doses, with a gradual decrease as the dose increased, but remaining at satisfactory levels. Conclusion Ionizing radiation promoted structural changes in the tetanus toxin such as fragmentation and/or aggregation and attenuation of enzymatic activity as the dose increased, but antigenic recognition of the toxin remained at good levels indicating its possible use as an immunogen. However, studies of enzymatic activity of tetanus toxin irradiated with doses above 8 kGy should be further analyzed.(AU)

Neuroactive venom compounds obtained from Phlogiellus bundokalbo as potential leads for neurodegenerative diseases: insights on their acetylcholinesterase and beta-secretase inhibitory activities in vitro

Spider venom is a rich cocktail of neuroactive compounds designed to prey capture and defense against predators that act on neuronal membrane proteins, in particular, acetylcholinesterases (AChE) that regulate synaptic transmission through acetylcholine (ACh) hydrolysis - an excitatory neurotransmitter - and beta-secretases (BACE) that primarily cleave amyloid precursor proteins (APP), which are, in turn, relevant in the structural integrity of neurons. The present study provides preliminary evidence on the therapeutic potential of Phlogiellus bundokalbo venom against neurodegenerative diseases. Methods Spider venom was extracted by electrostimulation and fractionated by reverse-phase high-performance liquid chromatography (RP-HPLC) and characterized by matrix-assisted laser desorption ionization-time flight mass spectrometry (MALDI-TOF-MS). Neuroactivity of the whole venom was observed by a neurobehavioral response from Terebrio molitor larvae in vivo and fractions were screened for their inhibitory activities against AChE and BACE in vitro. Results The whole venom from P. bundokalbo demonstrated neuroactivity by inducing excitatory movements from T. molitor for 15 min. Sixteen fractions collected produced diverse mass fragments from MALDI-TOF-MS ranging from 900-4500 Da. Eleven of sixteen fractions demonstrated AChE inhibitory activities with 14.34% (± 2.60e-4) to 62.05% (± 6.40e-5) compared with donepezil which has 86.34% (± 3.90e-5) inhibition (p > 0.05), while none of the fractions were observed to exhibit BACE inhibition. Furthermore, three potent fractions against AChE, F1, F3, and F16 displayed competitive and uncompetitive inhibitions compared to donepezil as the positive control. Conclusion The venom of P. bundokalbo contains compounds that demonstrate neuroactivity and anti-AChE activities in vitro, which could comprise possible therapeutic leads for the development of cholinergic compounds against neurological diseases.(AU)

Neuroactive venom compounds obtained from Phlogiellus bundokalbo as potential leads for neurodegenerative diseases: insights on their acetylcholinesterase and beta-secretase inhibitory activities in vitro

Spider venom is a rich cocktail of neuroactive compounds designed to prey capture and defense against predators that act on neuronal membrane proteins, in particular, acetylcholinesterases (AChE) that regulate synaptic transmission through acetylcholine (ACh) hydrolysis - an excitatory neurotransmitter - and beta-secretases (BACE) that primarily cleave amyloid precursor proteins (APP), which are, in turn, relevant in the structural integrity of neurons. The present study provides preliminary evidence on the therapeutic potential of Phlogiellus bundokalbo venom against neurodegenerative diseases. Methods Spider venom was extracted by electrostimulation and fractionated by reverse-phase high-performance liquid chromatography (RP-HPLC) and characterized by matrix-assisted laser desorption ionization-time flight mass spectrometry (MALDI-TOF-MS). Neuroactivity of the whole venom was observed by a neurobehavioral response from Terebrio molitor larvae in vivo and fractions were screened for their inhibitory activities against AChE and BACE in vitro. Results The whole venom from P. bundokalbo demonstrated neuroactivity by inducing excitatory movements from T. molitor for 15 min. Sixteen fractions collected produced diverse mass fragments from MALDI-TOF-MS ranging from 900-4500 Da. Eleven of sixteen fractions demonstrated AChE inhibitory activities with 14.34% (± 2.60e-4) to 62.05% (± 6.40e-5) compared with donepezil which has 86.34% (± 3.90e-5) inhibition (p > 0.05), while none of the fractions were observed to exhibit BACE inhibition. Furthermore, three potent fractions against AChE, F1, F3, and F16 displayed competitive and uncompetitive inhibitions compared to donepezil as the positive control. Conclusion The venom of P. bundokalbo contains compounds that demonstrate neuroactivity and anti-AChE activities in vitro, which could comprise possible therapeutic leads for the development of cholinergic compounds against neurological diseases.(AU)

Intoxicação de um canino por ingestão de Cannabis sativa / Intoxication of a canine by ingestion of Cannabis sativa

A Cannabis sativa, popularmente conhecida como "maconha", é uma planta que possui elevada solubilidade, absorção, distribuição e metabolização no organismo. Seu principal componente químico é a substância psicoativa delta-9-tetrahidrocanabiol e possui receptores com ampla distribuição anatômica, principalmente no sistema nervoso central, periférico e células do sistema imunológico, influenciando na ação de diferentes neurotransmissores e respostas imunomoduladoras. Seus efeitos dependem da quantidade de substância ingerida e os sinais clínicos acometem principalmente os sistemas neurológico, gastrointestinal e cardiovascular. O objetivo deste trabalho é relatar o caso de um canino, fêmea, sem raça definida, de dois meses de idade, acompanhada em uma clínica veterinária de Fortaleza/CE. O animal apresentava histórico de ingestão de maconha e sinais clínicos como ataxia, hiperestesia e desorientação, sendo assim, foi realizado o diagnóstico presuntivo baseado na anamnese, no exame físico e no laboratorial. Foi adotado um tratamento sintomático à base de fluidoterapia, furosemida e aspartato de L-Ornitina. O animal ficou internado durante 24h, apresentou boa resposta ao tratamento sem complicações secundárias, com redução dos sinais clínicos, e após isso recebeu alta.

Cannabis sativa, popularly known as "marijuana", is a plant that has high solubility, absorption, distribution and metabolization in the body. Its main chemical component is the psychoactive substance delta-9- tetrahydrocannabinol, which has receptors with wide anatomical distribution, mainly in the central, peripheral nervous system and cells of the immune system, influencing the action of different neurotransmitters and immunomodulatory responses. Its effects depend on the amount of substance ingested and the clinical signs mainly affect the neurological, gastrointestinal and cardiovascular systems. The objective of this work is to report the case of a canine, female, mixed breed, two months old, followed up at a veterinary clinic in Fortaleza / CE. The animal had a history of marijuana ingestion, and clinical signs such as ataxia, hyperesthesia and disorientation, thus making the presumptive diagnosis based on anamnesis, physical and laboratory examination. A symptomatic treatment based on fluid therapy, furosemide and L-Ornithine aspartate was adopted. The animal was hospitalized for 24 hours, presented a good response to treatment without secondary complications with reduced clinical signs, and after that he was discharged.

Neurobehavioral and neuroprotector effects of caffeine in animal models

This review aims to analyze and contrast the neurological effects associated with the use of caffeine on neurobehavior and neuroprotection in animal models. Caffeine belongs to the group of methylxanthines that exert a direct effect on adenosine receptors associated with inhibitory or excitatory G proteins, generating modification of cyclic AMP activity and intracellular calcium flow which produces alterations in the modulation system of the neurotransmitters dopamine and glutamate. The regulation of the neurotransmission systems generates protection against the inflammation of the central nervous system, by activation of the microglia and reinforcement of the blood-brain barrier. This drug will also restore cognition or prevent memory loss in Parkinson's or Alzheimer's diseases. It is important to establish new study models in other species to assess whether the behavior of the molecule is similar and to obtain other clinical applications in its behavioral and neuroprotective effects.(AU)

Neurobehavioral and neuroprotector effects of caffeine in animal models

This review aims to analyze and contrast the neurological effects associated with the use of caffeine on neurobehavior and neuroprotection in animal models. Caffeine belongs to the group of methylxanthines that exert a direct effect on adenosine receptors associated with inhibitory or excitatory G proteins, generating modification of cyclic AMP activity and intracellular calcium flow which produces alterations in the modulation system of the neurotransmitters dopamine and glutamate. The regulation of the neurotransmission systems generates protection against the inflammation of the central nervous system, by activation of the microglia and reinforcement of the blood-brain barrier. This drug will also restore cognition or prevent memory loss in Parkinson's or Alzheimer's diseases. It is important to establish new study models in other species to assess whether the behavior of the molecule is similar and to obtain other clinical applications in its behavioral and neuroprotective effects.

Neuroprotective properties of RT10, a fraction isolated from Parawixia bistriata spider venom, against excitotoxicity injury in neuron-glia cultures

L-Glutamate (L-Glu), the major excitatory neurotransmitter in the mammalian Central Nervous System (CNS), is essential to cognitive functions. However, when L-Glu is accumulated in large concentrations at the synaptic cleft, it can induce excitotoxicity that results in secondary damage implicated in many neurological disorders. Current therapies for the treatment of neurological disorders are ineffective and have side effects associated with their use; therefore, there is a need to develop novel treatments. In this regard, previous studies have shown that neuroactive compounds obtained from the venom of the spider Parawixia bistriata have neuroprotective effects in vitro and in vivo. In this sense, this work aimed to evaluate potential neuroprotective effects of fraction RT10, obtained from this spider venom, on primary cultures of neuron and glial cells subjected to glutamate excitotoxicity insults. Methods: Primary cultures of neurons and glia were obtained from the cerebral tissue of 1-day-old postnatal Wistar rats. After 7 days in vitro (DIV), the cultures were incubated with fraction RT10 (0.002; 0.02; 0.2 and 2 µg/µL) or riluzole (100 µM) for 3-hours before application of 5 mM L-Glu. After 12 hours, the resazurin sodium salt (RSS) test was applied to measure metabolic activity and proliferation of living cells, whereas immunocytochemistry for MAP2 was performed to measure neuronal survival. In addition, the cells were immunolabeled with NeuN and GFAP in baseline conditions. Results: In the RSS tests, we observed that pre-incubation with RT10 before the excitotoxic insults from L-Glu resulted in neuroprotection, shown by a 10% reduction in the cell death level. RT10 was more effective than riluzole, which resulted in a cell-death reduction of 5%. Moreover, qualitative analysis of neuronal morphology (by MAP2 staining, expressed as fluorescence intensity (FI), an indirect measure of neuronal survival) indicate that RT10 reduced the toxic effects of L-Glu, as shown by a 38 % increase in MAP2 fluorescence when compared to L-Glu insult. On the other hand, the riluzole treatment resulted in 17% increase of MAP2 fluorescence; therefore, the neuroprotection from RT10 was more efficacious. Conclusion: RT10 fraction exhibits neuroprotective effects against L-Glu excitotoxicity in neuron-glia cultured in vitro.(AU)

In silico analysis of binding interaction of conantokins with NMDA receptors for potential therapeutic use in Alzheimer's disease

Background The N-methyl-D-aspartate (NMDA) receptors are glutamate receptors that play vital roles in central nervous system development and are involved in synaptic plasticity, which is an essential process for learning and memory. The subunit N-methyl D-aspartate receptor subtype 2B (NR2B) is the chief excitatory neurotransmitter receptor in the mammalian brain. Disturbances in the neurotransmission mediated by the NMDA receptor are caused by its overexposure to glutamate neurotransmitter and can be treated by its binding to an antagonist. Among several antagonists, conantokins from cone snails are reported to bind to NMDA receptors. Methods This study was designed to analyze the binding mode of conantokins with NMDA receptors in both humans and rats. To study interactions, dockings were performed using AutoDock 4.2 and their results were further analyzed using various computational tools. Results Detailed analyses revealed that these ligands can bind to active site residues of both receptors as reported in previous studies. Conclusions In light of the present results, we suggest that these conantokins can act as antagonists of those receptors and play an important role in understanding the importance of inhibition of NMDA receptors for treatment of Alzheimer's disease.(AU)

In silico analysis of binding interaction of conantokins with NMDA receptors for potential therapeutic use in Alzheimer's disease

Background The N-methyl-D-aspartate (NMDA) receptors are glutamate receptors that play vital roles in central nervous system development and are involved in synaptic plasticity, which is an essential process for learning and memory. The subunit N-methyl D-aspartate receptor subtype 2B (NR2B) is the chief excitatory neurotransmitter receptor in the mammalian brain. Disturbances in the neurotransmission mediated by the NMDA receptor are caused by its overexposure to glutamate neurotransmitter and can be treated by its binding to an antagonist. Among several antagonists, conantokins from cone snails are reported to bind to NMDA receptors. Methods This study was designed to analyze the binding mode of conantokins with NMDA receptors in both humans and rats. To study interactions, dockings were performed using AutoDock 4.2 and their results were further analyzed using various computational tools. Results Detailed analyses revealed that these ligands can bind to active site residues of both receptors as reported in previous studies. Conclusions In light of the present results, we suggest that these conantokins can act as antagonists of those receptors and play an important role in understanding the importance of inhibition of NMDA receptors for treatment of Alzheimer's disease.(AU)

Estudos comportamentais e neurofisiológicos da exposição prolongada à cetamina como antidepressivo e o modelo de separação materna para indução de depressão pós-parto em ratos

A depressão pós-parto é uma doença multifatorial mentalmente incapacitante que afeta mundialmente as mulheres. O tratamento recomendado para essa enfermidade consiste na combinação de psicoterapia e tratamento farmacológico. Atualmente, a busca de antidepressivos está em expansão, sendo priorizados fármacos com mecanismo de ação misto. A cetamina vem sendo bastante estudada para o uso como antidepressivo, incluindo o tratamento para a depressão pós-parto. Apesar disso, há poucas informações na literatura a respeito da segurança do uso da cetamina durante o período pós-natal, tornando-se necessários mais estudos sobre o uso desse fármaco com essa finalidade. Assim, no presente trabalho propõe-se estudar os efeitos antidepressivos da exposição prolongada de cetamina em ratos adultos e em ratas lactantes induzidas à depressão pós-parto. Para tanto, o trabalho foi dividido em três capítulos. O Capítulo 1 tem como objetivo estudar os efeitos do modelo de separação materna para indução da depressão pós-parto em ratas. Para isso, ratas lactantes foram separadas de seus filhotes por 3 h diárias do dia pós-natal 2 ao 12. Foram realizados estudos comportamentais, neuroquímicos, hormonais, de expressão de RNA e de citocinas pró-inflamatórias. Os resultados mostraram que as ratas submetidas ao modelo de separação materna apresentaram alterações no comportamento materno, no nado forçado, nos níveis neurotransmissores cerebrais e nos níveis de corticosterona e TSH, consistentes com um fenótipo depressivo. Esses dados em conjunto sugerem que o modelo de separação materna em ratas pode ser útil para avaliação de fármacos com potencial atividade para o tratamento da depressão pós-parto. O Capítulo 2 tem como objetivo estudar os efeitos fármaco-toxicológicos do tratamento prolongado com diferentes doses subanestésicas de cetamina com potencial atividade antidepressiva em ratos. Para isso, ratos adultos foram tratados com 5, 10 ou 20 mg/kg de cetamina, por via intraperitoneal, por 21 dias e foi feita a avaliação do peso corporal, do consumo de água e ração e da bateria de observação funcional (FOB) semanalmente, bem como as análises neuroquímica, hematológica e bioquímica sérica ao final do tratamento. Os resultados mostraram alterações no ganho de peso e no sistema serotoninérgico. Em conjunto, esses dados indicam que as alterações observadas podem ser explicadas pelo mecanismo de ação da cetamina, sugerindo ausência de efeitos fármaco-toxicológicos desse fármaco nas doses utilizadas. O Capítulo 3 tem como objetivo estudar os efeitos do tratamento com diferentes doses subanestésicas de cetamina em ratas lactantes induzidas à depressão pós-parto. Para isso, ratas lactantes foram induzidas a depressão pós-parto pelo modelo descrito no Capítulo 1, tratadas com as mesmas doses de cetamina do Capítulo 2, durante toda a lactação, e submetidas as mesmas avaliações descritas no Capítulo 1. Os resultados mostraram que o tratamento prolongado com cetamina causou alterações no ganho de peso, no comportamento materno, na caixa claro/escuro, na neuroquímica cerebral e induziu comportamento estereotipado. Esses dados em conjunto sugerem que o tratamento prolongado com doses subanestésicas de cetamina não causou efeito antidepressivo em ratas induzidas à depressão pós-parto pelo modelo de separação materna, e que a dose mais alta foi capaz de induzir comportamento estereotipado.

Postpartum depression is a multifactorial mental illness that affects women worldwide. The approved treatment for this disease consists of a combination of psychotherapy and pharmacological treatment. Currently, there is growing interest in the development of new antidepressants, prioritizing drugs with mixed mechanism of action. Ketamine is being extensively studied for its use as an antidepressant, including as a treatment for postpartum depression. However, there is limited evidence on the safety of its use during the postnatal period. For this reason, further studies on the use of this drug as a treatment for postpartum depression during the postnatal period are needed. Thus, the present work aims to study the antidepressant effects of prolonged exposure to ketamine in adult rats and in lactating female rats that have been induced to postpartum depression. This work is divided into three chapters. Chapter 1 aims to study the effects of the maternal separation model for postpartum depression induction in female rats. For this purpose, dams were separated from their offspring for 3 hours per day between postnatal days 2 to 12. Behavioral, neurochemical, hormonal, RNA expression and pro-inflammatory cytokine studies were carried out. The results showed that the dams submitted to the maternal separation model showed changes in: maternal behavior, the forced swim test, brain neurotransmitters levels and corticosterone and TSH levels, consistent with a depressive phenotype. These data suggest that the maternal separation model in rats may be useful for evaluating drugs with potential activity for the treatment of postpartum depression. Chapter 2 aims to analyze the pharmaco-toxicological effects of prolonged treatment using different subanesthetic doses of ketamine with potential antidepressant activity in rats. Adult male rats were treated with 5, 10 or 20 mg/kg of ketamine, via intraperitoneal injections, for 21 days. Their body weight, water and food intake and the functional observation battery (FOB) were evaluated weekly. In addition, neurochemical, hematological and biochemical analyzes were conducted at the end of treatment. The results showed changes in weight gain and in the brain serotonergic system. Together, these data indicate that the observed changes can be explained by ketamine mechanism of action. This suggests that the doses used did not have any pharmaco-toxicological effects. Chapter 3 examines the effects of the treatment using different subanesthetic doses of ketamine in lactating rats induced to postpartum depression. For this purpose, dams were induced to postpartum depression by the same model described in Chapter 1, treated with the same doses of ketamine as in Chapter 2 throughout lactation, and submitted to the same assessments described in Chapter 1. The results showed that prolonged treatment using ketamine caused changes in weight gain, maternal behavior, the light/dark box test, the brain neurochemistry and induced stereotyped behavior. Taken together, these data suggest that prolonged treatment using subanesthetic doses of ketamine did not cause an antidepressant effect in dams induced to postpartum depression by the maternal separation model, whereas the higher dose of ketamine was able to induce stereotyped behavior.

A cetamina associada ou não ao álcool, quais as consequências toxicológicas e sua influência no estresse oxidativo? Estudo em ratos

O uso recreacional e os efeitos deletérios de drogas lícitas e/ou ilícitas é considerado um problema de saúde pública. Dentre estas drogas destaca-se a cetamina, um anestésico empregado na veterinária e o etanol, a droga lícita mais utilizada. Além dos efeitos centrais, estudos revelam que a cetamina e o etanol possuem propriedades imunomodulatórias. No entanto, poucos estudos são realizados com a associação destas drogas; assim, o objetivo deste estudo foi o de avaliar os efeitos tóxicos, imunotóxicos, o potencial oxidativo e os possíveis efeitos neurotóxicos resultantes do consumo associado ou não das mesmas. Para isso, ratos Wistar foram tratados uma vez ao dia, por até 28 dias, com injeções intraperitoneais (15 ou 30mg/kg/PV) de cetamina (K) ou com etanol a 10% (E) via oral (gavagem ou adicionado a água de bebida), ou ainda em associação (KE). O grupo controle recebeu apenas os veículos dos tratamentos. Ao final do experimento, os animais foram submetidos à eutanásia para realização de avaliações toxicológicas, imunológicas, oxidativas e antioxidativas, e de neurotransmissores centrais. Os resultados revelaram redução no ganho de peso dos animais tratados com KE e aumento de ácido ascórbico urinário. A redução de pH e glicose urinária foi observada nos grupos E e KE. Na bioquímica, todos os grupos apresentaram aumento de HDL, porém a associação das drogas levou a um aumento do colesterol, enquanto no grupo K, observou-se diminuição dos triglicérides e da fosfatase alcalina. Ainda, somente o grupo KE apresentou alterações na função renal. Todos os grupos experimentais exibiram alterações histopatológicas hepáticas e/ou vesicais. Os grupos E e KE apresentaram alterações no número de células mieloides e linfoides concomitantes ao aumento na celularidade de medula óssea. Apenas animais do grupo KE apresentaram alteração na resposta do tipo Th2. Foi observado o aumento da peroxidação lipídica nos animais tratados com K, bem como aumento na atividade de GPx e CAT e do conteúdo de GSSG dos animais tratados com E e/ou KE nos órgãos analisados. Ao nível central, observou-se elevação dos níveis de DA e NOR no hipocampo dos animais do grupo E, e 5HT nos animais do grupo K; além de aumento de DA e 5HT no córtex pré-frontal dos animais dos grupos E e K, respectivamente. Assim, concluímos que a associação KE promove redução do ganho de peso não relacionado ao consumo de ração. O etanol, em associação ou não, promove alterações nos parâmetros urinários; a cetamina promove diminuição nos níveis de FA e as drogas quando em associação alteram o perfil lipídico e renal na dependência da dose e do tempo de administração. Efeitos pró-oxidantes entraram em equilíbrio devido à ação de antioxidantes. O etanol, em associação ou não, promove alterações em células do sistema imune, no entanto, sem promover imunomodulação sobre suas respostas. O etanol e/ou a cetamina também promovem alterações histopatológicas hepáticas, enquanto que a associação ainda promove lesões vesicais. Além disso, ambas as drogas promovem alterações no perfil neuroquímico central; porém, quando associadas não promoveram efeitos sinérgicos sobre os parâmetros avaliados.

The recreational use and the deleterious effects of licit and/or illicit drugs are considered a public health issue. Among these drugs are ketamine, used in veterinary anaesthesia, and ethanol the most commonly licit drug. Besides their central effects, studies have shown that ketamine and ethanol have immunomodulatory properties. However, few studies are conducted with the association of both drugs; thus, we aimed to evaluate the toxic and immunotoxic effects, as well the oxidative potential and the possible neurotoxic effects resulted from the consumption of these drugs associated or not. For this, Wistar rats were treated once daily for up to 28 days with intraperitoneal injections (15 or 30mg/kg/BW) of ketamine (K) or orally with 10% ethanol (E) (gavage or drinking water) or the association of both treatments (KE). Control group received only vehicles. At the end of the experimental period, the animals were euthanized for toxicological, immunotoxicological, oxidative stress and antioxidant status evaluation, and central levels of neurotransmitters. KE animals showed reduced body weight gain and increased levels of urinary ascorbic acid. Reduction of urinary pH and glucose was observed in E and KE groups. Biochemistry analysis showed an increase in HDL levels in all experimental groups; although, KE showed an increase in cholesterol levels, while K group exhibited reduction in triglycerides and alkaline phosphatase (ALP) levels. Only KE group showed renal function alterations. All experimental groups showed hepatic and/or bladder histopathology alterations. E and KE groups exhibited alterations in myeloid and lymphoid cells number with concomitant increased in bone marrow cellularity. Only animals of KE group presented changes in Th2 response. Increased in lipid peroxidation was observed in K-treated animals, as well as GPx and CAT activities and GSSG content of animals treated with E and/or KE. We observed elevation in DA and NOR in hippocampus of E group and 5HT in K group, in addition to an increase in DA and 5HT of the prefrontal cortex of E and K groups, respectively. Thus, we conclude that KE association promotes reduction in body weight gain not related to food consumption. Ethanol, in combination or not, promotes urinary alterations; ketamine reduces ALP levels and the drugs, when in combination, alter lipid and renal profile depending on dose and time of administration. Pro-oxidant effects were balanced due to the action of antioxidants. Ethanol, associated or not, promotes alterations on immune cells without immunomodulatoty responses. Ethanol and/or ketamine also promotes liver histopathological alterations, while this association promotes bladder lesions. In addition, both drugs promote changes in central neurochemical levels; however, when associated do not promoted synergistic effects in evaluated parameters.

Behavioral and neurochemical evidence of deltamethrin anxiogenic-like effects in rats / Evidências neuroquímicas e comportamentais do efeito ansiogênico da deltametrina em ratos

Pyrethroid insecticides are extensively used for pest control around the house, flea prevention for pets, and plant sprays for the home and in agriculture. Deltamethrin (DTM) is a Type II pyrethroid insecticide used to control a variety of insects in agriculture and domestic environments. The present study investigated the possible anxiogenic effects of DTM (1, 3, and 10 mg/kg) in rats using behavioral and neurochemical methods. We assessed general locomotor activity and behavior in the elevated plus maze and open field test. Striatal and hippocampal neurotransmitter and metabolite levels were also measured. DTM (i) reduced locomotion and rearing frequency, (ii) slightly increased the duration of immobility, (iii) reduced the time engaged in social interaction, (iv) reduced the percentage of entries into and time spent on the open arms of the elevated plus maze, (v) reduced the number of center crossings in the elevated plus maze, (vi) Striatal and hippocampal neurotransmitter and metabolite levels were also measured. DTM (i) reduced locomotion and rearing frequency, (ii) slightly increased the duration of immobility, (iii) reduced the time engaged in social interaction, (iv) reduced the percentage of entries into and time spent on the open arms of the elevated plus maze, (v) reduced the number of center crossings in the elevated plus maze, (vi) increased striatal serotonin neurotransmitter and its metabolite, and (vii) did not alter motor coordination on the rotarod, grooming duration in the open field test, rectal temperature, or hippocampal neurotransmitter levels. These data suggest that DTM at the present doses and under these experimental conditions presented a similar profile to that of anxiogenic drugs, unrelated with the increased serotonin neurotransmission.(AU)

Inseticidas piretróides são amplamente utilizados para controle de pragas, como na prevenção de pulgas em animais de estimação e sprays de plantas para a casa e na agricultura. Deltametrina (DTM) é um inseticida piretróide tipo II usado para controlar uma variedade de insetos na agricultura e ambientes domésticos. O presente estudo investigou os possíveis efeitos ansiogênicos de DTM (1, 3 e 10 mg/kg) em ratos, utilizando métodos comportamentais e neuroquímicos. Foi avaliada a atividade locomotora geral e comportamento no labirinto em cruz elevado e teste de campo aberto. Os níveis de neurotransmissores e metabólitos no estriado e hipocampo também foram mensurados. DTM (i) reduziu a locomoção e a frequência de levantar, (ii) aumentou da duração da imobilidade, (iii) reduziu o tempo de interacção social, (iv) reduziu a percentagem de entradas e tempo gasto nos braços abertos do elevado labirinto em cruz, (v) reduziu o número de cruzamentos no centro do labirinto em cruz elevado, (vi) aumentou neurotransmissor serotonina e de seu metabólito estriatal, e (vii) não alterou a coordenação motora no rotarod, duração do grooming no teste de campo aberto, temperatura retal, ou níveis de neurotransmissores do hipocampo. Estes dados sugerem que DTM nas presentes doses e sob estas condições experimentais apresentaram um perfil semelhante ao de drogas ansiogénicas, não relacionados ao aumento da serotonina estriatal.(AU)

Repeated collection of cerebrospinal fluid from the lumbosacral region of ewes / Repeated collection of cerebrospinal fluid from the lumbosacral region of ewes

Background: Cerebrospinal fl uid is a vital fl uid from the central nervous system. Since CSF contains proteins, enzymes, hormones, neuropeptides and neurotransmitters that play critical regulatory roles in many different physiological processes, it has been extensively studied to explore different nervous disorders. Since CSF is a vital fl uid from the CNS, used for clinical examination of the CNS in ruminants and other domestic species. CSF may be collected in ewes under field conditions, which allows the diagnosis of bacterial and metabolic diseases, as well as using it for cytological studies and biochemical analysis. Depending on the study, in opportunity the sampling protocol should be repeated to measure dynamic changes in the parameters selected for the analysis. Under fi eld conditions, obtaining CSF samples from ewes is a diffi cult task. Thus, the aim of this work was to determine if it is possible to obtain repeated extractions of CSF by lumbosacral puncture from the same ewes under fi eld conditions.Materials, Methods & Results: The CSF was sampled in three successive weekly collections from nine ewes sedated with ketamine. The procedure collections were made by the same trained operator, who stood behind the ewe, facing its back. Having checked that the sagital plane of the animal was perpendicular to the horizontal plane the puncture point was found by manual

Background: Cerebrospinal fl uid is a vital fl uid from the central nervous system. Since CSF contains proteins, enzymes, hormones, neuropeptides and neurotransmitters that play critical regulatory roles in many different physiological processes, it has been extensively studied to explore different nervous disorders. Since CSF is a vital fl uid from the CNS, used for clinical examination of the CNS in ruminants and other domestic species. CSF may be collected in ewes under field conditions, which allows the diagnosis of bacterial and metabolic diseases, as well as using it for cytological studies and biochemical analysis. Depending on the study, in opportunity the sampling protocol should be repeated to measure dynamic changes in the parameters selected for the analysis. Under fi eld conditions, obtaining CSF samples from ewes is a diffi cult task. Thus, the aim of this work was to determine if it is possible to obtain repeated extractions of CSF by lumbosacral puncture from the same ewes under fi eld conditions.Materials, Methods & Results: The CSF was sampled in three successive weekly collections from nine ewes sedated with ketamine. The procedure collections were made by the same trained operator, who stood behind the ewe, facing its back. Having checked that the sagital plane of the animal was perpendicular to the horizontal plane the puncture point was found by manual

Efeitos da sibutramina e do metilfenidato em modelo animal do Transtorno do Déficit de Atenção e Hiperatividade (TDAH) induzido em camundongos por etanol no período pós-natal

O transtorno do déficit de atenção e hiperatividade (TDAH) é uma condição que pode ser caracterizada pela falta de atenção, impulsividade e hiperatividade. A fisiopatologia do TDAH está relacionada, principalmente, a alterações no sistema dopaminérgico, noradrenérgico e serotoninérgico do sistema nervoso central. Dentre os tratamentos utilizados destaca-se a farmacoterapia com metilfenidato, potencial droga de abuso, que age como inibidor da recaptação de dopamina, noradrenalina e serotonina; por outro lado, o sal de sibutramina monoidratada, que possui mecanismo de ação farmacológico semelhante nestes sistemas de neurotransmissão central, ainda não teve sua utilização testada em um modelo do TDAH. Assim, o objetivo deste trabalho foi estudar os efeitos da administração prolongada (28 32 dias) de sibutramina e de metilfenidato em modelo animal do TDAH induzido pela exposição ao etanol no período pós-natal em camundongos, avaliando-se o ganho de peso semanal, o consumo de água e de ração, bem como o comportamento animal, por meio da avaliação geral no campo aberto e nos testes do labirinto em cruz elevado, da suspensão pela cauda, do reconhecimento de objetos e do labirinto em T. Foram avaliados também os níveis dos neurotransmissores e seus metabólitos em diferentes estruturas cerebrais. Os resultados mostraram que o modelo animal do TDAH induzido pela exposição ao etanol no período pós-natal apresentou hipoatividade no campo aberto seguida de aumento da atividade, não apresentou alterações nos níveis de ansiedade no labirinto em cruz elevado, como também mostrou comportamento tipo-depressivo no teste de suspensão pela cauda e marcante déficit na memória de trabalho e atenção no teste de reconhecimento de objetos e labirinto em T. Em relação ao tratamento prolongado com sibutramina e metilfenidato, não foram observadas alterações no ganho de peso semanal e consumo de água e ração. No campo aberto o metilfenidato normalizou a atividade dos camundongos, enquanto a sibutramina causou hiperatividade. No labirinto em cruz elevado não foram observadas alterações nos níveis de ansiedade. No teste de suspensão pela cauda o metilfenidato ocasionou comportamento tipo-depressivo nos camundongos salina, enquanto a sibutramina reverteu os efeitos depressivos dos etanol. O metilfenidato melhorou a memória de trabalho e atenção dos camundongos que receberam etanol tanto no teste de reconhecimento de objetos quanto no labirinto em T, já a sibutramina foi capaz de fazê-lo apenas no labirinto em T.

The attention deficit hyperactivity disorder (ADHD) is a condition that can be characterized by the lack of attention, impulsivity and hyperactivity. The pathophysiology of ADHD is related mainly to changes in the dopaminergic system, noradrenergic and serotoninergic of central nervous system. Among the treatments used stands out the pharmacotherapy with methylphenidate, potential drug of abuse, which acts as an inhibitor of the reuptake of dopamine, noradrenaline and serotonin; on the other hand, the salt of sibutramine monohydrate, which has a pharmacological mechanism of action similar in these systems of central neurotransmission, have not had their use tested in a model of ADHD. Thus, the objective of this work was to study the effects of prolonged administration (28 - 32 days) of sibutramine and methylphenidate in an animal model of ADHD induced by exposure to ethanol in the postnatal period in mice, evaluating the weight gain weekly, the consumption of water and feed, as well as animal behavior, through the general assessment in open field, and in the elevated plus maze, and in the tests of tail suspension, the recognition of objects and the T maze. We evaluated the levels of neurotransmitters and their metabolites in different brain structures. The results showed that the animal model of ADHD induced by exposure to ethanol in the postnatal period showed hypoactivity in the open field followed by increased activity, showed no changes in the levels of anxiety in the elevated plus maze, as also shown depressive-like behavior in the tail suspension test and striking deficit in working memory and attention in the test of recognition of objects and T maze. In relation to the prolonged treatment with sibutramine and methylphenidate, no alterations were observed in weight gain weekly, and consumption of water and food. In the open field, methylphenidate normalized the activity of mice, while sibutramine caused hyperactivity. In the elevated plus maze no changes were observed in anxiety levels. In the tail suspension test methylphenidate caused depressive-like behavior in mice salina, while sibutramine reversed the effects of depression of ethanol. Methylphenidate has improved the working memory and attention of mice that received ethanol both in the test of recognition of objects as the maze in T, since sibutramine was able to do it only in the maze in T.

Repeated collection of cerebrospinal fluid from the lumbosacral region of ewes

Background: Cerebrospinal fluid is a vital fluid from the central nervous system. Since CSF contains proteins, enzymes, hormones, neuropeptides and neurotransmitters that play critical regulatory roles in many different physiological processes, it has been extensively studied to explore different nervous disorders. Since CSF is a vital fluid from the CNS, used for clinical examination of the CNS in ruminants and other domestic species. CSF may be collected in ewes under field conditions, which allows the diagnosis of bacterial and metabolic diseases, as well as using it for cytological studies and biochemical analysis. Depending on the study, in opportunity the sampling protocol should be repeated to measure dynamic changes in the parameters selected for the analysis. Under field conditions, obtaining CSF samples from ewes is a difficult task. Thus, the aim of this work was to determine if it is possible to obtain repeated extractions of CSF by lumbosacral puncture from the same ewes under fi eld conditions. Materials, Methods & Results: The CSF was sampled in three successive weekly collections from nine ewes sedated with ketamine. The procedure collections were made by the same trained operator, who stood behind the ewe, facing its back. Having checked that the sagital plane of the animal was perpendicular to the horizontal plane the puncture point was found by manual palpation at the slight depression between the ends of the spinous apophyses of the last lumbar and first sacral vertebrae. The wool was separated, and the area was cleansed with iodine solution. The puncture was performed with a spinal needle, after it had penetrated through the skin, the needle was pushed forward very slowly. When was listening for any vibrations ('clicks'), suggesting that the needle had crossed the dural membrane and entered into the arachnoidal space. Then, the syringe needle was withdrawn and the CSF came out slowly, either immediately or after some slow movements of the needle. If CSF did not come out, the puncture was deepened further on until the ventral arachnoidal space was reached. In the first and second collection, limpid CSF samples were obtained in all (9/9, 100%) and in 8/9 animals (89%), respectively. However, limpid CSF samples were obtained only in 4 of the animals one week later (4/9, 44 % P = 0.01). The volume of CSF extracted ranged from 0.6 to 0.9 mL/sample/animal. Discussion: The sequential collection of CSF in ewes is possible under field conditions to obtain a high percentage of samples to the along of three weekly extraction events. When only the first extraction event was considered, the sampling was totally effective even entirety of the animals. Yet by the third sampling, we obtained fewer samples than in the second event. In the present technique of repeated puncture was yielded a high efficacy in the first collection at random chosen ewes. The decrease in effectiveness was probably due to cumulative tissue damage and formation of extensive fibrous adhesions in the subarachnoidal space, which would compromise partially or totally the flow of CSF. The volume of CSF collected by ewe along the three repeated extractions did not vary, although it tended to decrease as repeated collections were performed. This tendency could also be linked with cumulative tissue damage. Nevertheless, our range of volume of CSF obtained for ewe is similar to volumes obtained in similar report. We concluded that the efficiency of weekly CSF extraction in ewes managed under field conditions decreases in the third sampling occasion.

ATIVIDADE CITOTÓXICA E ANTI-INFLAMATÓRIA DE EXTRATO DE FOLHAS DE Jatropha curcas L. EM CÉLULAS GLIAIS

A Jatropha curcas L. (J. curcas L.), planta da família da Euforbeáceae, conhecida localmente como pinhão manso, apresenta alta plasticidade adaptativa, sendo cultivada em diferentes regiões de clima tropical e subtropical do mundo. Configura-se uma alternativa na medicina popular tradicional e veterinária para tratamentos de várias condições patológicas. Estudos demonstram que ação farmacológica como antimicrobiano, antifúngico, antioxidante, cicatrizante e anti-inflamatório possa está associado à presença de classes de compostos bioativos como os flavonoides e taninos. No entanto, não existem relatos da ação de compostos isolados desta planta no Sistema Nervoso Central (SNC). A escassez de conhecimento técnico-científico sobre a espécie gera uma demanda por estudos mais aprofundados. Assim o presente estudo objetivou realizar um mapeamento prospectivo sitemático sobre evolução tecnológica das diversas utilizações da planta Jatropha curcas L., obter extrato metanólico de folhas de J. curcas L. (EJC) e suas frações (FnEJC), realizar triagem fitoquímica qualitativa dos bioativos do EJC e investigar a atividade citotóxica e anti-inflamatória em culturas primárias de células gliais submetidas a estímulo inflamatório. A prospecção de registros tecnológicos a partir da base de dados do Escritório Europeu de Patentes (Espacenet®), entre o período de 1983 a 2012, não revelou documentos de tecnologiais (patentes) com finalidade terapêutica farmacológica da J. curcas L. A triagem fitoquímica do EJC apresentou resultado positivo para xantonas, leucoantocianidinas, catequinas (taninos catéquicos), flavanonas e flavonoides, xantonas, esteroides, terpenoides e saponinas. Foi observada atividade anti-inflamatória do extrato metanólico de folhas de J. curcas L. e frações em células gliais obtidas a partir de córtex cerebral de ratos Wistar neonatos, tratadas com lipopolissacarideo de Escherichia coli (LPS 1 g mL-1). O EJC (0,1 g mL-1 a 1.000 g mL-1) não foi citotóxica para as células gliais e apresentou EC50 de 10,794 g mL-1. O tratamento com LPS induziu ativação de astrócitos e microglias caracterizada por modificações morfológicas e da expressão de marcadores epecíficos GFAP e Iba-1, respectivamente além dada ativação do Fator Nuclear kaap B (NF-kB) em astrócitos além de aumento da produção de Oxido nítrico (NO). O Tratamento com o EJC preservou células quiescentes e atenuou ativação de células gliais pós indução inflamatória. Observou-se que EJC, F1EJC e F2EJC (0,1 g mL-1) inibiram a produção de NO, inclusive após estímulo inflamatório com LPS. Os resultados sugerem que o EJC inibe a ativação do NF-kB por apresentar compostos bioativos da J. curcas L. com possível bloqueio da resposta glial na regulação de genes envolvidos no processo inflamatório. Os resultados também contribuíram para a geração do documento de depósito de patente no Instituto Nacional da Propriedade Industrial (INPI) no intuito de assegurar as novas descobertas desenvolvidas.

Jatropha curcas L. (Euforbeáceae), known locally as pinhão manso, is a plant with highly adaptable plant cultivated in different regions of tropical and subtropical regions of the world. Its adopeted in traditional folk medicine for the treatment of various pathological conditions, including arthritis, wounds and especially inflammation. Parts of the plant heve been demonstrated biological activites, as antimicrobial, antifungal, antioxidant, healing and anti-inflammatort, however, there are no evidences about effects on the Central Nervous System (CNS). Increasing evidence shows that glia cells, in speciall astrocytes and microglia, play a crucial role in the inflammatory response in the CNS, implicated in the pathogenesis and progression of neurodegenerative diseases. The glial reaction (gliosis) is chacterized by a set of alterations including changes in the synthesis and distribution of cytoskeletal proteins, release of molecules involved in inflammation, and reduced ability to regulate extracellular neurotransmitters and ions. The objective of the present work was to perform a prospective study about the technological evolution of the various uses of the Jatropha curcas L., to obtain a methanolic extract from leaves of J. curcas L. (EJC) and its fractions (FnEJC), to perform qualitative phytochemical study of EJC and to investigate the cytotoxic and anti-inflammatory activity in primary cultures of glial cells submitted to inflammatory stimuli. The prospection of technological registrations from the database of the European Patent Office (Espacenet®), the platform with the largest number of associates in the world, between the yars of 1983 and 2012, did not reveal technology documents (patents) with pharmacological therapeutic purpose of J. curcas L. Phytochemical screening of EJC showed positive results for xanthones, leucoanthocyanidins, catechins (catheteric tannins), flavanones and flavonoids, xanthones, steroids, terpenoids and saponins. Glial cell cultures were obtained from the cerebral cortex of neonates Wistar rats and treated wit LPS (1 g mL-1). EJC at 0.1 1,000 g mL-1 was not toxic to glial cells, and presented EC50 of 10.794 g mL-1. Treatment with LPS induced astrocyte and microglia activation characterized by morphological modifications and changes on expression of GFAP and Iba-1, markers of astrocytes and microglia, respectively, as well as activation of transcription factor NF-kB and production of NO. EJC treatment associated with LPS attenuated gliosis and activation of the LPS-induced Nuclear kaap B Factor (NF-kB). Moreover, (EJC) and fractions F1EJC and F2EJC reduced LPS-induced prodution of NO. Togheter these results indicates that EJC presentes anti-inflammatory antivity in reative glial cells associated to inibition of NF-kB. The results also contributed to the generation of the patent document of invention in the National Institute of Industrial Property (INPI) in order to ensure the new discoveries developed.