Efeito do ?cido asc?rbico sobre a espermatog?nese de camundongos com distrofia muscular de Duchenne

Autor: Braz, Janine Karla Fran?a da Silva
Jazyk: portugalština
Rok vydání: 2015
Předmět:
Zdroj: Repositório Institucional da UFRNUniversidade Federal do Rio Grande do NorteUFRN.
Druh dokumentu: masterThesis
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Objetivou-se avaliar as altera??es na espermatog?nese provocadas pela Distrofia Muscular de Duchenne (DMD) e o efeito do tratamento com ?cido asc?rbico na preven??o dessas inj?rias. Neste trabalho foram utilizados 24 camundongos, sendo 12 linhagem C57BL/10 (n?o-distr?ficos) e 12 C57BL/10Mdx (distr?ficos) divididos em seis grupos com 4 animais cada (C30 = Controle de 30 dias; D30 = Distr?fico com 30 dias; C60 = Controle com 60 dias; D60 = Distr?fico com 60 dias; CS = Controle com 60 dias suplementados com ?cido asc?rbico e; DS60 = Distr?fico com 60 dias suplementados com ?cido asc?rbico. Os animais dos grupos C30 e D30 foram eutanasiados aos trinta dias de idade, enquanto que os animais dos demais grupos aos 60 dias de idade. A suplementa??o com ?cido asc?rbico foi ministrada na ?gua na dosagem de 0,005g/dia durante 30 dias. Ap?s eutanasiados, os test?culos (direito e esquerdo) foram coletados, imediatamente, pesados e seccionados transversalmente, fixados em solu??o de Karnovysky, inclu?dos em resina histol?gica (an?lises morfol?gicas e morfom?tricas), submetidos a an?lise ultraestrutural e submentidos t?cnica de imunohistoqu?mica para caspase-3. Houve aumento significativo no percentual de t?nica pr?pria em D30 em rela??o a C30 e D60. As an?lises ultraestruturais indicaram ind?cios de apoptose mitocondrial das c?lulas de Sertoli que podem reduzir a efici?ncia esperm?tica em CS60 e DS60. Verificou-se maior densidade de volume das c?lulas apopt?ticas postivas para Caspase - 3 em D30 versus C30 e DS60 em rela??o a CS60.. Houve acentuada hipertrofia de c?lulas de Leydig entre D30 e D60. No entanto, com a suplementa??o observou-se revers?o dessa altera??o em DS60. Na ultraestrutura das c?lulas de Leydig observou-se a presen?a precoce de ves?culas lip?dicas no grupo distr?fico pr?-p?bere (D30). Dessa forma, a DMD afetou a organiza??o dos t?bulos semin?feros e intert?bulos, no entanto, a suplementa??o de ?cido asc?rbico., nas condi??es experimentais utilizadas para o tratamento da DMD foi suficiente apenas para reduzir a hipertofia das c?lulas de Leydig.
The objective was to evaluate spermatogenesis alterations caused by DMD and the effect of the treatment using ascorbic acid in preventing those injuries. Twenty four mice were used, 12 from the C57BL/10 lineage (non-dystrophic) and 12 from the C57BL/10Mdx (dystrophic). The sample was divided in six groups containing 4 animals each, as: C30 = 30 days control; D30 = Dystrophic with 30 days; C60 = 60 days control; D60 = Distrophic with 60 days; CS60 = 60 days control supplemented with ascorbic acid and DS60 = Dystrophic with 60 days supplemented with ascorbic acid. The ascorbic acid supplementation was given in the water, 0,005 mg/day. After euthanasia, the testicles (right and left) were collected, weighted and cross sectioned. The material was fixed in the Karnovsky solution for 24 hours, included in resin for histological studies (morphological and morphometric analyzes) submitted to ultrastructural analysis and immunohistochemistry for caspase-3. There was a significant increase in the tunica propria percentage in D30 compared to C30 and D60. The ultrastructural analysis showed mitochondrial apoptosis evidence of Sertoli cells that can reduce sperm efficiency in CS60 and DS60. A higher volume density of apoptotic cells postivas to Caspase-3 in C30 and D30 versus DS60 compared to CS60. There was severe hypertrophy of the Leydig cells between D30 and D60. However, with supplementation was observed reversal of this change in DS60. The ultrastructure of Leydig cells to early presence of lipid vesicles was observed in the group pre-pubertal dystrophic (D30). Thus, the DMD affect the organization of the seminiferous tubules and intertubule, however, the ascorbic acid supplementation used for the treatment of DMD has been just enough to reduce the hypertrophy of the Leydig cells.
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