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Previous issue date: 2013-03-22 / Epilepsy affects 1% of the world population and 30% of these patients are refractory to available medication. Stem cells host hope in the treatment of epilepsy. Given their ability to proliferate, differentiate and production of factors which may activate endogenous mechanisms to restore the injured brain. Knowing that the administration of bone marrow mononuclear cells (BMMC) in animals have therapeutic potential in an experimental model of epilepsy, the aim of this study is to investigate the mechanisms by which administered cells exert their beneficial. In order to better understand the mechanisms of action of transplanted cells, a comparative study was done to detect the expression of trophic factors as brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), nerve growth factor (NGF), transforming growth factor beta (TGF-R) and vascular endothelial growth factor (VEGF) in hippocampi of each experimental groups by ELISA. Experimental model of epilepsy was induced by pilocarpine injection (320 mg/kg; ip). Seizures were scored by Racine s scale. The duration of SE was controlled with diazepan (10mg/kg; ip; 90 minutes after SE onset). Twenty-two days after SE, rats were randomly assigned into groups: Control, Pilo, Pilo+BMMC evaluated in periods 3, 7 and 14 days after transplant. BMMC groups received cell transplantation (obtained from EGFP C57BL/6 mice) via tail vein (1x107 cells, 100L). While control animals received saline instead of pilocarpine. Pilocarpine-treated animals were monitored for the presence of spontaneous seizures for 22 days (7 days prior to cell transplant). Our results showed that there was a change in the protein expression of BDNF, GDNF, NGF, TGF-R and VEGF in the hippocampus of epileptic animals treated with BMMC compared to untreated epileptic animals and control, with variations in the expression of each factor at different times after transplantation. The expression of BDNF, GDNF, NGF and VEGF was high and TGF-R1 reduced after transplantation of BMMC compared to untreated epileptic animals. However, there was no difference in the expression of these factors in untreated epileptic animals compared to control animals, except TGF-R1, which proved to be high in the group of untreated epileptic animals compared to control animals. The results of this study provide additional data on the potential benefit of BMMC as well as provide insight into the mechanism by which BMMC promote functional recovery in epileptic rats. / A epilepsia atinge cerca de 1% da popula??o mundial, sendo que aproximadamente 30% desses pacientes n?o respondem ao tratamento medicamentoso. Por sua vez, as c?lulas-tronco t?m sido consideradas uma esperan?a de tratamento da epilepsia, visto que t?m grande capacidade de prolifera??o, diferencia??o e produ??o de fatores, podendo ativar mecanismos de restaura??o end?gena no c?rebro lesado. Sabendo-se que a administra??o de c?lulas mononucleares da medula ?ssea (CMMO) apresenta potencial terap?utico em um modelo experimental de epilepsia, o objetivo deste estudo ? investigar se o transplante das CMMO em ratos com epilepsia cr?nica modula a express?o de fatores tr?ficos. Com o intuito de melhor compreender os mecanismos de a??o das c?lulas transplantas foi realizada a detec??o da express?o de fatores tr?ficos como o fator neurotr?fico derivado do c?rebro (BDNF), fator neurotr?fico derivado da glia (GDNF), fator de crescimento neural (NGF), fator de crescimento transformador R1 (TGF-R1) e fator de crescimento endotelial vascular (VEGF) em diferentes per?odos ap?s transplante das CMMO nos hipocampos dos grupos experimentais atrav?s da t?cnica de ELISA. A pilocarpina (PILO) foi administrada nos animais (320 mg/kg i.p.,) para indu??o do modelo de epilepsia cr?nica. As crises comportamentais foram classificadas de acordo com a escala de Racine e a dura??o do SE foi controlada com diazepam (10 mg/kg, i.p., 90 minutos). Ap?s 22 dias, o total dos animais foi dividido em grupos: Controle, Pilo e Pilo+CMMO avaliados nos tempos de 3, 7 e 14 dias ap?s o transplante. Os grupos CMMO receberam transplante de c?lulas da camada mononuclear da medula ?ssea, obtidas de camundongos EGFP C57BL/6, via veia caudal (1x107 c?lulas, 100L). Os animais controle receberam solu??o salina nas mesmas condi??es do grupo transplantado. Os animais tratados com pilocarpina foram v?deo-monitorados durante sete dias pr?-transplante para observa??o de crises espont?neas recorrentes (CERs). Nossos resultados mostraram que houve altera??o da express?o proteica de BDNF, GDNF, NGF, TGF-R1 e VEGF nos hipocampos dos animais epil?pticos tratados com as CMMO em rela??o aos animais epil?pticos n?o tratados e controle, havendo varia??es da express?o de cada fator em diferentes tempos ap?s o transplante. A express?o dos fatores BDNF, GDNF, NGF e VEGF mostrou-se elevada e do TGF-R1 reduzida ap?s o transplante das CMMO em rela??o aos animais epil?pticos n?o tratados. Por?m, n?o houve diferen?a na express?o destes fatores nos animais epil?pticos n?o tratados em rela??o aos animais controle, exceto o TGF-R1, o qual se mostrou elevado no grupo de animais epil?pticos n?o tratados em rela??o aos animais controle. Os resultados deste estudo fornecem dados adicionais sobre o benef?cio do potencial das CMMO, bem como fornecer uma vis?o sobre o mecanismo pelo qual as CMMO favorecem a recupera??o funcional em ratos epil?pticos.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/1723 |
| Date | 22 March 2013 |
| Creators | Zanirati, Gabriele Goulart |
| Contributors | Costa, Jaderson Costa da |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Medicina e Ci?ncias da Sa?de, 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/masterThesis |
| 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 | 7620745074616285884, 500, 600, -8624664729441623247 |
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