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Previous issue date: 2013-03-01 / Objectives: to investigate separately the therapeutic potential of neuropeptide NAP and the intra-arterial (IA) transplantation of human umbilical cord blood (HUCB) mononuclear cells in an animal model of neonatal hypoxia-ischemia (HI). Methods: male Wistar rats at postnatal day 7 were subjected to an HI model by permanent occlusion of right common carotid artery and systemic hypoxia (8% O2 for 2 h). The animals were randomly assigned to groups receiving an intraperitoneal injection of NAP (3 μg/g) immediately (0 h) and 24 h after HI. Other animals received HUCB mononuclear cells (1 x 106 or 1 x 107 cells/50 μL) into the left common carotid artery 24 h after HI insult by using the microneedle technique. Only for NAP study, brain DNA damage, lipid peroxidation and reduced glutathione (GSH) content were determined 48 h post-HI insult. The spatial version of the Morris water maze learning task and the stereological volume assessment were performed in adult animals that received both therapies. The accelerated rotarod test and cerebral and body weight determination were applied only for cellular therapy study. The HUCB mononuclear cells migration was monitored through nested-PCR analysis in the brain and systemic organs of transplanted-HI rats. Results: we observed that NAP prevented the acute HI-induced DNA and lipid membrane damage and also recovered the GSH levels in the injured hemisphere of the HI rat pups. Further, NAP was able to hinder impairments in learning and long-term spatial memory and to significantly reduce brain damage in adult animals previously subjected to neonatal HI. The IA transplantation in neonatal HI rat seemed to be a feasible and safe delivery route for HUCB mononuclear cells. The intra-arterially delivered cells hindered dose-dependently the learning and spatial memory impairments without brain damage recovery in adult rats. Additionally we further showed that HI insult or IA cell transplantation had no long-term impact in the body weight and motor function in rodents. The HUCB mononuclear cells could be promptly identified in the ischemic brain after IA transplantation and also in some peripheral organs until at least 30 days later. Conclusions: the viability and long-lasting beneficial effects of neuropeptide NAP and IA transplantation of HUCB mononuclear cells support its translational characteristic for neonatal HI management. Therefore, NAP and intracarotid delivery of cells became promising candidates for the treatment of HI-induced brain damage and life-long disabilities / Objetivos: investigar, separadamente, o potencial terap?utico do neuropept?deo NAP e do transplante intra-arterial de c?lulas mononucleares de sangue de cord?o umbilical humano (CMSCUH) em um modelo experimental de hip?xia-isquemia (HI) neonatal. M?todos: ratos Wistar machos com 7 dias de vida foram submetidos ao modelo de HI neonatal atrav?s da oclus?o permanente da art?ria car?tida comum direita e hip?xia sist?mica (8% O2 por 2 h). A administra??o do neuropept?deo NAP (3 μg/g, i.p.) ocorreu imediatamente (0 h) e 24 h ap?s a aplica??o do modelo. J? o transplante de CMSCUH (1 x 106 ou 1 x 107 c?lulas/50 μL) foi realizado 24 h p?s-HI atrav?s de t?cnica de microagulha na art?ria car?tida comum esquerda. Somente no estudo do NAP, ensaio cometa alcalino, peroxida??o lip?dica e n?veis de glutationa reduzida foram determinados no hipocampo e c?rtex cerebral 48 h p?s-HI. Para ambas as terapias empregaram-se a vers?o espacial do labirinto aqu?tico de Morris (LAM) e determina??o estereol?gica da volumetria hemisf?rica cerebral na fase adulta destes animais. Utilizou-se o teste do rotarod acelerado e pesagem corporal e cerebral dos animais adultos apenas no estudo do transplante intra-arterial de CMSCUH. A migra??o das CMSCUH foi monitorada atrav?s da t?cnica de nested-PCR no c?rebro e ?rg?os sist?micos. Resultados: o tratamento com NAP restabeleceu a integridade hipocampal e cortical do DNA e membranas lip?dicas, junto ao incremento do sistema glutationa. Al?m disso, o NAP impediu o desenvolvimento de d?ficits do aprendizado e da mem?ria espacial juntamente ? redu??o da atrofia cerebral em ratos adultos. O transplante intracarot?dico de CMSCHU em ratos neonatos HI demonstrou-se fact?vel e seguro. Este tratamento preveniu, de maneira dose-dependente, o desenvolvimento de preju?zo cognitivo de ratos adultos previamente expostos ? HI neonatal, sem influ?ncia sobre a atrofia cerebral. A fun??o motora e peso corporal de ratos adultos n?o sofreram influ?ncia da HI neonatal ou da administra??o intra-arterial de CMSCHU. As CMSCHU puderam ser detectadas tanto logo ap?s o transplante como tardiamente (30 dias) no c?rebro e ?rg?os sist?micos dos animais transplantados. Conclus?es: os efeitos ben?ficos e duradouros do neuropept?deo NAP e do transplante intra-arterial de CMSCHU, assim como sua viabilidade, fornecem evid?ncias de uma poss?vel translacionalidade destas duas terapias inovadoras para o tratamento da HI neonatal. Desta forma, o neuropept?deo NAP e a via intracarot?dica para o transplante de CMSCHU tornam-se candidatos potenciais para a preven??o do dano HI cerebral e decorrentes sequelas
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/1410 |
| Date | 01 March 2013 |
| Creators | Greggio, Samuel |
| Contributors | Costa, Jaderson Costa da |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Medicina/Pediatria e Sa?de da Crian?a, PUCRS, BR, Faculdade de Medicina |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da PUC_RS, instname:Pontifícia Universidade Católica do Rio Grande do Sul, instacron:PUC_RS |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 3098206005268432148, 500, 600, -8624664729441623247 |
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