Submitted by PPG Medicina e Ci?ncias da Sa?de (medicina-pg@pucrs.br) on 2018-03-22T14:21:52Z No. of bitstreams: 1 RODRIGO_BRACCINI_MADEIRA_DA_SILVA_TES.pdf: 6388000 bytes, checksum: c5e864eb62bd553c3a29e7bad1b85fef (MD5) / Approved for entry into archive by Tatiana Lopes (tatiana.lopes@pucrs.br) on 2018-04-05T12:24:56Z (GMT) No. of bitstreams: 1 RODRIGO_BRACCINI_MADEIRA_DA_SILVA_TES.pdf: 6388000 bytes, checksum: c5e864eb62bd553c3a29e7bad1b85fef (MD5) / Made available in DSpace on 2018-04-05T12:31:22Z (GMT). No. of bitstreams: 1 RODRIGO_BRACCINI_MADEIRA_DA_SILVA_TES.pdf: 6388000 bytes, checksum: c5e864eb62bd553c3a29e7bad1b85fef (MD5) Previous issue date: 2018-01-16 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Voltage-gated calcium channels (VGCC) play a critical role in neuroinflammatory diseases, such as multiple sclerosis (MS). We investigated the effects of the recombinant peptide CTK 01512-2, an inhibitor of N-type VGCC/TRPA1-mediated calcium influx, in the mouse model of experimental autoimmune encephalomyelitis (EAE). The effects of this molecule were compared to those displayed by ziconotide - a selective N-type VGCC blocker clinically used for chronic pain, and fingolimod - an orally active drug employed for MS treatment. The intrathecal (i.t.) administration of CTK 01512-2 markedly prevented hyperalgesia, body weight loss, splenomegaly, MS-like clinical and neurological scores, impaired motor coordination and spatial memory, with an efficacy comparable to that observed for ziconotide and fingolimod. This molecule displayed a favourable profile on EAE-induced neuroinflammatory changes, including inflammatory infiltrate, demyelination, increased pro-inflammatory cytokines, glial activation and glucose hypermetabolism in brain and spinal cord. The recovery of spatial memory, besides a reduction of serum leptin levels, allied to central and peripheral elevation of the anti-inflammatory cytokine IL-10, were solely modulated by CTK 01512-2, dosed intrathecally. The systemic i.v. administration of CTK 01512-2 also reduced the EAE-elicited MS-like symptoms and signals, similarly to that seen in animals that received fingolimod orally. Ziconotide lacked any significant effect when dosed by i.v. route. Collectively, these results indicate that CTK 01512-2 greatly improved the neuroinflammatory responses in a mouse model of MS, with a higher efficacy when compared to ziconotide, pointing out this molecule as a promising adjuvant for MS management. / Os canais de c?lcio voltagem-dependentes (CCVD) desempenham um papel importante nas doen?as neuroinflamat?rias, como na esclerose m?ltipla (EM). Foram investigados os efeitos do tratamento com o pept?deo recombinante, CTK 01512-2, um bloqueador dos CCVD do tipo N/TRPA1 - respons?veis pelo influxo de c?lcio, no modelo animal de encefalomielite autoimune experimental (EAE) em camundongos. Os efeitos dessa mol?cula foram comparados aos exibidos pela ziconotida ? um bloqueador seletivo dos CCVD do tipo N, usado clinicamente para dor cr?nica. As a??es do CTK 01512-2 tamb?m foram comparadas ?quelas exercidas pelo fingolimode ? um f?rmaco utilizado por via oral para o tratamento da EM. A administra??o intratecal (i.t.) de CTK 01512-2 preveniu marcadamente a hiperalgesia, perda de peso corporal, esplenomegalia, escores cl?nicos e neurol?gicos relacionados a EAE, comprometimento na coordena??o motora e mem?ria espacial, com efic?cia compar?vel ? observada para ziconotida e fingolimode. Essa mol?cula mostrou um perfil favor?vel nas mudan?as neuroinflamat?rias induzidas por EAE, incluindo infiltrado inflamat?rio, desmieliniza??o, aumento de citocinas pr?-inflamat?rias, ativa??o glial e no metabolismo da glicose no c?rebro e medula espinhal. A recupera??o da mem?ria espacial, redu??o dos n?veis s?ricos de leptina, al?m do aumento dos n?veis centrais e perif?ricos da citocina anti-inflamat?ria, IL-10, foram modulados apenas pelo CTK 01512-2, quando administrado pela via i.t. A administra??o sist?mica (i.v.) de CTK 01512-2 tamb?m reduziu os sinais e sintomas evocados por EAE, de forma semelhante ao fingolimode, administrado por via oral. A ziconotida n?o teve efeito significativo quando administrada pela via i.v. Coletivamente, esses resultados indicam que o CTK 01512-2 melhorou significativamente a resposta neuroinflamat?ria no modelo de animal de EM em camundongos, com efic?cia superior quando comparado com a ziconotida, apontando essa mol?cula como um adjuvante promissor no tratamento da EM.