Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-12-04T20:41:48Z No. of bitstreams: 1 JoaquimGoncalvesDeAzevedoNeto_DISSERT.pdf: 1209476 bytes, checksum: b176b29c8fe26fd7b3e020bb5279ef38 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-12-07T19:42:32Z (GMT) No. of bitstreams: 1 JoaquimGoncalvesDeAzevedoNeto_DISSERT.pdf: 1209476 bytes, checksum: b176b29c8fe26fd7b3e020bb5279ef38 (MD5) / Made available in DSpace on 2017-12-07T19:42:32Z (GMT). No. of bitstreams: 1 JoaquimGoncalvesDeAzevedoNeto_DISSERT.pdf: 1209476 bytes, checksum: b176b29c8fe26fd7b3e020bb5279ef38 (MD5) Previous issue date: 2017-04-27 / Nociceptina-orfanina FQ (N/OFQ) ? um heptadecapept?deo, que atua como ligante end?geno do receptor NOP. O receptor NOP tem ampla express?o no sistema nervoso e sua ativa??o induz efeitos inibit?rios, causando redu??o da libera??o de neurotransmissores e/ou inibi??o do disparo neuronal, dependendo do s?tio (pr? ou p?s-sin?ptico) no qual ? expresso. Estudos in vivo t?m demonstrado o envolvimento do sistema N/OFQ-receptor NOP na modula??o dos estados emocionais. Este estudo visou avaliar os efeitos do agonista NOP, Ro65-6570, sobre as altera??es comportamentais de camundongos submetidos a uma situa??o de estresse incontrol?vel. Para tanto, foi utilizado o modelo do desamparo aprendido (DA), que emprega eletrochoques, incontrol?veis e imprevis?veis, nas patas dos animais como evento estressor para induzir um fen?tipo do tipo desamparado, que consiste num d?ficit do comportamento de escape do compartimento eletrificado. Nem todos os animais expostos ao DA desenvolvem o fen?tipo do tipo desamparado, logo, dois grupos distintos foram observados: camundongos desamparados e n?o-desamparados. Ap?s a exposi??o ao DA, foram realizados testes para observar as altera??es comportamentais que o estresse causou, como labirinto em cruz elevado (LCE), nata??o for?ada e campo aberto. Foi observado efeito ansiog?nico do DA nos camundongos desamparados, com diminui??o no n?mero de entradas e tempo gasto nos bra?os abertos do LCE, quando comparados aos grupos n?o-desamparados e naive. Ap?s essa etapa, foi avaliada a a??o do diazepam (1 mg/kg, via i.p.) e do agonista NOP, Ro65-6570 (1 mg/kg, via i.p.), no comportamento do tipo ansioso dos animais desamparados, bem como em sua locomo??o. O diazepam foi capaz de reverter o comportamento do tipo ansioso observado no teste do LCE pelos camundongos desamparados, sem preju?zo na locomo??o dos animais. De forma similar, a administra??o de Ro65-6570 tamb?m apresentou efeito ansiol?tico somente nos camundongos desamparados, sem afetar a atividade locomotora. Em conclus?o, este estudo aponta o sistema da N/OFQ-receptor NOP como um alvo terap?utico inovador para o tratamento de transtornos psiqui?tricos relacionados ao estresse. / Nociceptin/orphanin FQ (N/OFQ) is a heptadecapeptide, acting as an endogenous ligand of the NOP receptor. The NOP receptor has wide expression in the central nervous system and its activation induces inhibitory effects, causing reduction of neurotransmitters release and/or inhibition of neuronal firing, depending on the site (pre or post-synaptic) in which it is expressed. In vivo studies have showed the involvement of the N/OFQ-NOP receptor system in the modulation of emotional states. This study aimed to evaluate the effects of the NOP agonist, Ro65-6570, on the mouse behavioral changes after an uncontrollable stressful situation due to unpredictable electric footshocks. To achieve this aim, the learned helplessness model (LH) was used as a stressor to induce a helpless behavior. Helpless phenotype consists of a deficit in the escape behavior of the electrified compartment. Not all animals exposed to the LH developed the helpless phenotype, and then two distinct groups were observed: helpless and non-helpless mice. After the LH, tests were performed aimed to identify the behavioral changes induced by stress, such as elevated plus maze (EPM), forced swimming and open field. LH exposure induced anxiogenic-like effects in helpless mice. In fact, when compared to non-helpless and naive controls, helpless mice displayed decreased number of entries and time spent in the open arms of the EPM. After then, the effects of diazepam (1 mg/kg, via ip) and the NOP agonist, Ro65-6570 (1 mg/kg, via ip) were assessed in the anxiogenic-like behavior of helpless mice, as well as in spontaneous locomotion. Diazepam was able to reverse the anxiogenic-related behaviors of helpless mice in the EPM, without affecting locomotion. Similarly, the administration of Ro65-6570 induced anxiolytic effects only in helpless mice, without changes in the locomotor activity. In conclusion, this study supports to the N/OFQ-NOP receptor system as an innovative therapeutic target for the treatment of psychiatric disorders related to stress.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-05-31T20:39:36Z No. of bitstreams: 1 IrisUcellaDeMedeiros_TESE.pdf: 1627038 bytes, checksum: abacbae1a6fa2f0ce88c897b540dedc4 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-06-03T00:28:56Z (GMT) No. of bitstreams: 1 IrisUcellaDeMedeiros_TESE.pdf: 1627038 bytes, checksum: abacbae1a6fa2f0ce88c897b540dedc4 (MD5) / Made available in DSpace on 2016-06-03T00:28:56Z (GMT). No. of bitstreams: 1 IrisUcellaDeMedeiros_TESE.pdf: 1627038 bytes, checksum: abacbae1a6fa2f0ce88c897b540dedc4 (MD5) Previous issue date: 2015-08-14 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Durante as ?ltimas d?cadas t?m sido demonstrado que existe uma rela??o entre a depress?o maior e a ativa??o do sistema imunol?gico. Nociceptina/orfanina FQ (N/OFQ) ? o ligante natural do receptor acoplado ? prote?na Gi chamado NOP, ambos constituem um sistema pept?dico que est? envolvido na regula??o do humor e de respostas inflamat?rias. Considerando essas a??es, a presente tese teve como objetivo investigar as consequ?ncias do bloqueio da sinaliza??o do receptor NOP nos comportamentos doentio e do tipo depressivo induzidos pela administra??o de lipopolissacar?deo (LPS) em camundongos. A administra??o sist?mica de doses de LPS, que n?o causam sepse, induzem altera??es nos comportamentos de camundongos relacionadas com a atividade das citocinas pr?-inflamat?rias fator de necrose tumoral-? (TNF-?) e interleucinas 6 (IL-6) e 1? (IL- 1 ?). Ap?s 2 a 6 h e 24 h da inje??o intraperitoneal, camundongos tratados com LPS apresentam, respectivamente, o comportamento doentio e o comportamento do tipo depressivo. No presente trabalho, a administra??o de LPS (0,8 mg/kg, ip) induziu sinais de doen?a em camundongos Swiss e CD-1, como perda de peso, redu??o transit?ria da temperatura retal e diminui??o da ingest?o de ra??o e ?gua. Al?m disso, nas 24 h ap?s a inje??o de LPS, essas mesmas linhagens de camundongos mostraram aumento no tempo de imobilidade durante os 6 min que foram submetidos ao teste de suspens?o pela cauda (TSC). O tratamento com nortriptilina (30 mg/kg, ip, 60 minutos antes do TSC) reduziu o tempo de imobilidade dos camundongos controle e tratados com LPS, e foi utilizado como antidepressivo padr?o. A administra??o pr?via ao LPS do antagonista do receptor NOP SB-612111 (10 mg/kg, ip), n?o alterou os comportamento doentio e do tipo depressivo induzidos pelatoxina. No entanto, quando injetado 24 h ap?s o tratamento com LPS, SB-612111 (ip, 30 minutos antes do TSC), como tamb?m o antagonista pept?dico do receptor NOP UFP-101 (10 nmol/2ul, icv, 5 min antes do TSC), inverteram significativamente os efeitos da toxina. O protocolo de indu??o do comportamento do tipo depressivo pela administra??o intraperitoneal de LPS tamb?m foi testado em camundongos nocautes para o receptor NOP (NOP(-/-)) e seus respectivos wild types (NOP(+/+)). O tratamento com LPS provocou altera??es significativas, como a redu??o tempor?ria da temperatura retal nos camundongos NOP(-/-) e perda de peso corporal e redu??o no consumo de ra??o e ?gua em ambos os camundongos NOP(+/+) e NOP(-/-). O consumo de ?gua foi significativamente diferente entre os gen?tipos. A inje??o de LPS induziu altera??es transit?rias nas citocinas pr?-inflamat?rias. Nas 6 horas ap?s o tratamento com LPS, os n?veis s?ricos de TNF-? mostraram-se aumentados significativamente nos camundongos NOP (+/+) e NOP (-/-), j? os n?veis de IL-6 mostraram-se significativamente aumentados apenas no soro dos camundongos NOP (+/+). Nas 24 horas ap?s a inje??o de LPS, os n?veis s?ricos das citocinas pr?-inflamat?rias retornaram ? linha de base. O tratamento com LPS provocou efeitos de tipo depressivo nos camundongos NOP (+/+), mas foi ineficaz nos camundongos NOP (-/-). Os dados obtidos nesta tese mostram que o bloqueio farmacol?gico e gen?tico da sinaliza??o mediada pelo receptor NOP n?o previne os comportamentos doentio e do tipo depressivo induzidos por LPS, no entanto revertem o comportamento do tipo depressivo. Em conclus?o, esses resultados evidenciam o envolvimento do sistema pept?dico N/OFQ - receptor NOP na modula??o dos comportamentos relacionados ao humor e ? ativa??o do sistema imunol?gico. / During the last decades, it has been established that there is a relationship between major depression and activation of immune system. Nociceptin/orphanin FQ (N/OFQ) is the natural ligand of a Gi-protein coupled receptor named NOP, both compose the peptidergic system wich is involved in the regulation of mood states and inflammatory responses. Considering these actions, the present thesis aimed to investigate the consequences of blocking NOP signaling in lipopolysaccharide (LPS)-induced sickness and depressive-like behaviors in mice. Systemic administration of LPS doses, that do not cause sepsis in mice, induce changes in their behaviors related with activity of pro-inflammatory cytokines tumor necrosis factor-? (TNF-?) and interleukins 6 (IL-6) and 1? (IL-1 ?). At the time points of 2 to 6 h and 24 h after intraperitoneal injection, mice treated with LPS displayed, respectively, sickness and depressive-like behaviors. In the present work the administration of LPS 0.8 mg/kg (ip) significantly induced sickness signs in Swiss and CD-1 mice, such as weight loss, transient reduction in rectal temperature and decrease of food and water intake. Moreover at 24 h after LPS injection these same mice strains displayed significantly increased immobility time on the tail suspension test (TST) when compared with control mice, this alteration was not related with possible locomotion impairments as verified on the open field test. Treatment with Nortriptyline 30 mg/kg (ip, 60 min prior the TST) reduced the immobility time of control and LPS-treated mice and was used as standard antidepressant. The NOP receptor antagonist SB-612111 (10 mg/kg, ip), 30 min prior LPS, did not modify LPS-induced sickness signs and depressive-like behavior. However, when injected 24 h after LPS treatment, SB-612111 (ip, 30 min prior the TST) as well as the peptidergic NOP receptor antagonist UFP-101 (10 nmol/2?L, icv, 5 min prior the TST) significantly reversed the toxin effects. The protocol of LPS-induced depressive-like states was also tested in NOP receptor knockout mice (NOP(-/-)) and their respective wild types (NOP(+/+)). LPS evoked transient rectal temperature reduction in NOP(-/-) mice and loss of body weight, food and water intake reduction in both NOP(+/+) and NOP(-/-) mice. The consumption of water was significantly different due to the genotype. LPS injection induced transient changes in pro-inflammatory cytokines. At 6 h after LPS injection, serum levels of TNF-? were significantly increased in NOP(+/+) and NOP(-/-) mice, as the IL-6 levels were significantly increased just in NOP(+/+) serum. At 24 h after LPS treatment the pro-inflammatory cytokines had returned to the baseline levels in both genotypes. LPS treatment elicited depressive-like effects in NOP(+/+) but not in NOP(-/-) mice. The data obtained during the execution of this doctoral thesis reveal that pharmacological and genetic blockade of NOP signaling does not affect LPS evoked sickness signs while reversing depressive-like behavior. In conclusion, these results highlight the involvement of the peptidergic system N/OFQ - NOP receptor in the modulation of behaviors related to mood and activation of the immune system.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-06-02T23:17:43Z No. of bitstreams: 1 EpifanioFernandesDaSilva_DISSERT.pdf: 1808656 bytes, checksum: a66e4e7210681ca2462da4ceef086da5 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-06-12T21:13:37Z (GMT) No. of bitstreams: 1 EpifanioFernandesDaSilva_DISSERT.pdf: 1808656 bytes, checksum: a66e4e7210681ca2462da4ceef086da5 (MD5) / Made available in DSpace on 2017-06-12T21:13:37Z (GMT). No. of bitstreams: 1 EpifanioFernandesDaSilva_DISSERT.pdf: 1808656 bytes, checksum: a66e4e7210681ca2462da4ceef086da5 (MD5) Previous issue date: 2017-02-23 / INTRODU??O: A agressividade ? um comportamento comum a diversas esp?cies animais, incluindo humanos. Entretanto, a viol?ncia e a impulsividade, associadas ? agressividade, s?o um problema social e podem ser consideradas patol?gicas, pois est?o presentes em v?rios transtornos psiqui?tricos. Diversas ?reas encef?licas est?o associadas ? express?o do comportamento agressivo, como a am?gdala, hipot?lamo e c?rtex pr?-frontal. V?rios sistemas de neurotransmiss?o est?o mediando o comportamento agressivo, dentre eles: serotonina, dopamina, noradrenalina e GABA. De maneira geral, os alvos terap?uticos dispon?veis para controle da agressividade modulam a fun??o dos sistemas de neurotransmissores acima. A nociceptina/orfanina FQ (N/OFQ) ? um heptadecapept?deo que atua como ligante do receptor NOP. Evid?ncias cl?nicas e pr?-cl?nicas mostram o envolvimento do sistema N/OFQ ? receptor NOP com transtornos psiqui?tricos, incluindo aqueles nos quais a agressividade est? associada. OBJETIVO: Este trabalho investigou o efeito de f?rmacos cl?ssicos e ligantes do receptor NOP no comportamento agressivo de camundongos machos, por meio do teste do residente-intruso. M?TODOS: Foram utilizados camundongos Swiss machos. Valproato 300 mg/kg, L?tio 50 mg/kg, Carbamazepina 20 mg/kg e Diazepam 1 mg/kg foram os f?rmacos cl?ssicos utilizados nesse estudo. Dentre os ligantes NOP utilizados destacam-se: Ro 65-6570 (0,01 ? 1 mg/kg), agonista pleno, AT-090 (0,01-0,1 mg/kg), agonista parcial, SB-612111 (1- 10 mg/kg), antagonista NOP. Para o teste do residente-intruso, camundongos machos foram isolados por 7 dias (residentes). Nos 8? e 11? dia, foram realizadas sess?es de avalia??o da agressividade, por meio da inser??o de um camundongo intruso na caixa do residente por 10 min. No dia 8, a agressividade basal foi avaliada sem qualquer tratamento pr?vio; no dia 11, o mesmo camundongo residente foi novamente avaliado, ap?s ter recebido o tratamento relativo ao seu grupo experimental. O campo aberto foi utilizado para avaliar o efeito dos f?rmacos na atividade locomotora. RESULTADOS: Valproato, L?tio, Carbamazepina reduziram o comportamento agressivo no teste do residente-intruso, enquanto que o tratamento com Diazepam n?o afetou a agressividade dos residentes. A administra??o de Ro 65-6570 (em todas as doses testadas) e AT-090 (na dose mais alta), aumentou o comportamento agressivo. J? o agonista parcial, AT-090, nas menores doses, reduziu discretamente a agressividade dos residentes. O tratamento com SB-612111 n?o modificou o comportamento agressivo dos animais. Nenhum dos tratamentos alterou a atividade locomotora dos animais. CONCLUS?O: Os f?rmacos cl?ssicos utilizados na cl?nica para tratamento de transtornos psiqui?tricos, os quais incluem sintomas de agressividade, foram eficazes em controlar a agressividade nos camunodngos residentes. Por outro lado, a ativa??o do receptor NOP tende a aumentar o comportamento agressivo, enquanto que o bloqueio deste sinal n?o foi modifica este comportamento. Em ?ltima an?lise, com estes dados sugere-se que os agonistas NOP poderiam promover como efeito adverso aumento da agressividade. / INTRODUCTION: Several species including humans display aggressive behavior. However, violence and impulsivity related to aggressiveness represent a social problem. Indeed, aggressive behavior can be considered symptoms of many psychiatric disorders. Some of the brain areas involved in aggression include amygdala, hypothalamus, and prefrontal cortex. Aggressiveness is modulated by different neurotransmitters, such as serotonin, dopamine, noradrenaline and GABA. These systems represent the therapeutic targets available to treat aggressiveness. The nociceptin/orphanin FQ (N/OFQ) is a heptadecapeptide acting as endogenous ligand of NOP receptor. Clinical and preclinical findings suggest the involvement of N/OFQ ? NOP receptor system with psychiatric disorders, including those related to aggressiveness. AIM: This study investigated the effects of standard drugs as well as NOP receptor ligands on aggressiveness in mice submitted to the resident-intruder test. METHODS: Male Swiss mice were used to develop this study. Valproate 300 mg/kg, Lithium 50 mg/kg, Carbamazepine 20 mg/kg, and Diazepam 1 mg/kg were used as standard drugs. The NOP ligands Ro 65-6570 (0.01 ? 1 mg/kg), full agonist, AT-090 (0,01 ? 0,1 mg/kg), partial agonist, and SB-612111 (1 ? 10 mg/kg), antagonist, were used. In the resident-intruder test, male mice were housed individually for 7 days (residents) before the experiment. The aggressiveness of each resident mouse was tested twice, at 8th and 11th days, by inserting an intruder mouse in the resident cage for 10 min. Day 8 of experiment, the basal aggressiveness of resident mice was recorded without pharmacological treatment; Day 11 of experiment, the same mouse was re-tested after being treated. The open field was used to evaluated the spontaneous locomotor activity . RESULTS: Valproate, Lithium, and Carbamazepine reduced the aggressive behavior of resident mice, while Diazepam did not affect the agressiveness. Ro 65-6570 (at all doses) and AT-090 (at the highest dose), increased aggressiveness. The partial agonist, AT-090, at lowest doses, slightly reduced aggressive behavior. The treatment with SB-61211 did not modified the aggressive behavior of mice. None of the treatments affected the locomotor activity. CONCLUSION: Standard drugs used in therapy for psychiatric disorders were effective on aggressiveness control in the resident mice. In contrast, the activation of NOP receptor tends to increase the aggressive behavior, while the blockade of this signal did not modify this behavior. Ultimately, these data suggest that NOP agonists could increase aggressive behavior as an adverse event.