Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro

Detalhes bibliográficos
Ano de defesa: 2021
Autor(a) principal: Azevedo, Venância Antonia Nunes
Orientador(a): Batista, Ana Liza Paz Souza
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/60208
Resumo: The aim of this study was to investigate the effect of different concentrations of Aloe vera extract on growth, morphology, viability, antrum formation and mRNA levels for SOD, CAT, GPX1 and PRDX6 in vitro cultured bovine secondary follicles. For this, the ovaries of multiparous cows were obtained from a local slaughterhouse. In the laboratory, the ovarian cortex was fragmented, and the secondary follicles (150 to 250µm) were isolated and cultured individually at 38.5°C, CO2 at 5%, for 18 days in a control medium consisting of TCM-199 supplemented with BSA, insulin, transferrin, selenium, FSH, penicillin and streptomycin, ascorbic acid, glutamine and hypoxanthine. The treatments consisted of adding 2.5, 5.0, 10.0 or 20.0% of the Aloe vera extract to the culture medium. Every 6 days, Follicular growth (diameter and daily growth), morphology and antrum formation was evaluated. At the end of the culture, follicular viability (fluorescence with calcein / ethidium), and the expression of transcripts for antioxidant enzymes SOD, CAT, GPX1 and PRDX6, by real-time PCR, were evaluated. Data on follicular growth and daily growth rate were submitted to the Kolmogorov-Smirnov test, followed by the Kruskal-Wallis test. To analyze survival and antrum formation, the Chi-square test was used. The Student's t-test was used to assess the levels of luminous intensity for Calceina-AM and homodimer- ethidium. The mRNA levels were analyzed by the Kruskal-Wallis test. Differences were considered significant when P <0.05. The results showed that Aloe vera maintained follicular morphology similar to the control group. Follicles cultured the presence of Aloe vera showed a progressive increase in follicular diameter until the 12 day of culture. An increase in daily growth rate was observed from day 0 to day 6 in treatment with 5.0% Aloe vera when compared to the control group. Follicles cultured with 2.5% Aloe vera had higher rate of antrum formation than those cultured in the control group. Regarding viability, follicles cultured with 2.5 and 5.0% Aloe vera showed a higher viability rate than follicles cultured in control medium. The presence of 2.5, 5.0 and 20.0% Aloe vera in the culture medium maintained the levels of mRNA for SOD, CAT and PRDX6 in follicles similar to the control group. In conclusion, Aloe vera maintains the morphology of secondary follicles cultured in vitro for 18 days, and 2.5% of Aloe vera promotes antrum formation, increases follicular viability and maintains mRNA expression for SOD and PRDX6 after cultivation.
id UFC-7_85ee8b3ebc7b64107e4d951e6d986867
oai_identifier_str oai:repositorio.ufc.br:riufc/60208
network_acronym_str UFC-7
network_name_str Repositório Institucional da Universidade Federal do Ceará (UFC)
repository_id_str
spelling Azevedo, Venância Antonia NunesBatista, Ana Liza Paz Souza2021-09-02T00:33:42Z2021-09-02T00:33:42Z2021-05-18Azevedo, V.A.N. Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro. 2021. 105f. Dissertação (Mestrado em Biotecnologia) - Programa de Pós-Graduação em Biotecnologia, Campus de Sobral, Universidade Federal do Ceará, Sobral, 2021.http://www.repositorio.ufc.br/handle/riufc/60208The aim of this study was to investigate the effect of different concentrations of Aloe vera extract on growth, morphology, viability, antrum formation and mRNA levels for SOD, CAT, GPX1 and PRDX6 in vitro cultured bovine secondary follicles. For this, the ovaries of multiparous cows were obtained from a local slaughterhouse. In the laboratory, the ovarian cortex was fragmented, and the secondary follicles (150 to 250µm) were isolated and cultured individually at 38.5°C, CO2 at 5%, for 18 days in a control medium consisting of TCM-199 supplemented with BSA, insulin, transferrin, selenium, FSH, penicillin and streptomycin, ascorbic acid, glutamine and hypoxanthine. The treatments consisted of adding 2.5, 5.0, 10.0 or 20.0% of the Aloe vera extract to the culture medium. Every 6 days, Follicular growth (diameter and daily growth), morphology and antrum formation was evaluated. At the end of the culture, follicular viability (fluorescence with calcein / ethidium), and the expression of transcripts for antioxidant enzymes SOD, CAT, GPX1 and PRDX6, by real-time PCR, were evaluated. Data on follicular growth and daily growth rate were submitted to the Kolmogorov-Smirnov test, followed by the Kruskal-Wallis test. To analyze survival and antrum formation, the Chi-square test was used. The Student's t-test was used to assess the levels of luminous intensity for Calceina-AM and homodimer- ethidium. The mRNA levels were analyzed by the Kruskal-Wallis test. Differences were considered significant when P <0.05. The results showed that Aloe vera maintained follicular morphology similar to the control group. Follicles cultured the presence of Aloe vera showed a progressive increase in follicular diameter until the 12 day of culture. An increase in daily growth rate was observed from day 0 to day 6 in treatment with 5.0% Aloe vera when compared to the control group. Follicles cultured with 2.5% Aloe vera had higher rate of antrum formation than those cultured in the control group. Regarding viability, follicles cultured with 2.5 and 5.0% Aloe vera showed a higher viability rate than follicles cultured in control medium. The presence of 2.5, 5.0 and 20.0% Aloe vera in the culture medium maintained the levels of mRNA for SOD, CAT and PRDX6 in follicles similar to the control group. In conclusion, Aloe vera maintains the morphology of secondary follicles cultured in vitro for 18 days, and 2.5% of Aloe vera promotes antrum formation, increases follicular viability and maintains mRNA expression for SOD and PRDX6 after cultivation.O objetivo deste estudo foi investigar o efeito de diferentes concentrações do extrato de Aloe vera no crescimento, morfologia, viabilidade, formação de antro e os níveis RNAm para SOD, CAT, GPX1 e PRDX6 em folículos secundários bovinos cultivados in vitro. Para isso, os ovários de vacas multíparas foram obtidos em um abatedouro local. No laboratório, o córtex ovariano foi fragmentando e os folículos secundários (150 a 250µm) foram isolados e cultivados individualmente a 38,5°C, CO2 a 5%, por 18 dias em meio controle constituído pelo TCM-199 suplementado com BSA, insulina, transferrina, selênio, FSH, penicilina e estreptomicina, ácido ascórbico, glutamina e hipoxantina. Os tratamentos consistiram da adição de 2,5, 5, 10 ou 20% do extrato de Aloe vera ao meio de cultivo. A cada 6 dias, avaliou-se o crescimento folicular (diâmetro e crescimento diário), a morfologia e a formação do antro. Ao final do cultivo, a viabilidade folicular (fluorescência com calceína/etídeo) e a expressão de transcritos para enzimas antioxidantes SOD, CAT, GPX1 e PRDX6, por PCR em tempo real, foram avaliadas. Os dados referentes ao crescimento folicular e da taxa de crescimento diária foram submetidos ao teste de Kolmogorov- Smirnov, seguido do teste Kruskal-Wallis. Para analisar a sobrevivência e formação de antro foi utilizado o teste Qui-quadrado. O teste t de Student foi utilizado para avaliar os níveis de intensidade luminosa para CalceÍna- AM e Homodímero-etídio. Os níveis de mRNA foram analisados pelo teste de Kruskal-Wallis, seguido do teste post hoc não paramétrico. As diferenças foram consideradas significativas quando P <0,05. Os resultados mostraram que o Aloe vera manteve a morfologia folicular semelhante ao grupo controle. Os folículos cultivados na presença de Aloe vera apresentaram um aumento progressivo do diâmetro folicular até o dia 12 de cultivo. Um aumento na taxa de crescimento diário foi observado do dia 0 ao dia 6 no tratamento com 5% de Aloe vera quando comparado ao grupo de controle. Folículos cultivados com 2,5% de Aloe vera tiveram maior taxa de formação de antro do que aqueles cultivados no grupo controle. Quanto à viabilidade, folículos cultivados com 2,5 e 5% de Aloe vera apresentaram maior taxa de viabilidade do que folículos cultivados em meio controle. A presença de 2,5, 5 e 20% de Aloe vera no meio de cultura manteve os níveis de mRNA para SOD, CAT e PRDX6 nos folículos semelhantes ao grupo controle. Em conclusão, a Aloe vera mantém a morfologia de folículos secundários cultivados in vitro por 18 dias, sendo que 2,5% de Aloe vera promove a formação de antro, aumenta a viabilidade folicular e mantém a expressão de mRNA para SOD e PRDX6 após o cultivo.Aloe veraFoliculogêneseVacaEfeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitroEffect of Aloe Vera extract on growth, viability and gene expression of in vitro cultured bovine secondary folliclesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2021_dis_vanazevedo.pdf2021_dis_vanazevedo.pdfAzevedo, V.A.N. Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro. 2021. 105f. Dissertação (Mestrado em Biotecnologia) - Programa de Pós-Graduação em Biotecnologia, Campus de Sobral, Universidade Federal do Ceará, Sobral, 2021.application/pdf1950319http://repositorio.ufc.br/bitstream/riufc/60208/1/2021_dis_vanazevedo.pdfb881ca0d88c73321a9785279541e63dfMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/60208/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52riufc/602082021-09-01 21:33:42.969oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2021-09-02T00:33:42Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro
dc.title.en.pt_BR.fl_str_mv Effect of Aloe Vera extract on growth, viability and gene expression of in vitro cultured bovine secondary follicles
title Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro
spellingShingle Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro
Azevedo, Venância Antonia Nunes
Aloe vera
Foliculogênese
Vaca
title_short Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro
title_full Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro
title_fullStr Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro
title_full_unstemmed Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro
title_sort Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro
author Azevedo, Venância Antonia Nunes
author_facet Azevedo, Venância Antonia Nunes
author_role author
dc.contributor.author.fl_str_mv Azevedo, Venância Antonia Nunes
dc.contributor.advisor1.fl_str_mv Batista, Ana Liza Paz Souza
contributor_str_mv Batista, Ana Liza Paz Souza
dc.subject.por.fl_str_mv Aloe vera
Foliculogênese
Vaca
topic Aloe vera
Foliculogênese
Vaca
description The aim of this study was to investigate the effect of different concentrations of Aloe vera extract on growth, morphology, viability, antrum formation and mRNA levels for SOD, CAT, GPX1 and PRDX6 in vitro cultured bovine secondary follicles. For this, the ovaries of multiparous cows were obtained from a local slaughterhouse. In the laboratory, the ovarian cortex was fragmented, and the secondary follicles (150 to 250µm) were isolated and cultured individually at 38.5°C, CO2 at 5%, for 18 days in a control medium consisting of TCM-199 supplemented with BSA, insulin, transferrin, selenium, FSH, penicillin and streptomycin, ascorbic acid, glutamine and hypoxanthine. The treatments consisted of adding 2.5, 5.0, 10.0 or 20.0% of the Aloe vera extract to the culture medium. Every 6 days, Follicular growth (diameter and daily growth), morphology and antrum formation was evaluated. At the end of the culture, follicular viability (fluorescence with calcein / ethidium), and the expression of transcripts for antioxidant enzymes SOD, CAT, GPX1 and PRDX6, by real-time PCR, were evaluated. Data on follicular growth and daily growth rate were submitted to the Kolmogorov-Smirnov test, followed by the Kruskal-Wallis test. To analyze survival and antrum formation, the Chi-square test was used. The Student's t-test was used to assess the levels of luminous intensity for Calceina-AM and homodimer- ethidium. The mRNA levels were analyzed by the Kruskal-Wallis test. Differences were considered significant when P <0.05. The results showed that Aloe vera maintained follicular morphology similar to the control group. Follicles cultured the presence of Aloe vera showed a progressive increase in follicular diameter until the 12 day of culture. An increase in daily growth rate was observed from day 0 to day 6 in treatment with 5.0% Aloe vera when compared to the control group. Follicles cultured with 2.5% Aloe vera had higher rate of antrum formation than those cultured in the control group. Regarding viability, follicles cultured with 2.5 and 5.0% Aloe vera showed a higher viability rate than follicles cultured in control medium. The presence of 2.5, 5.0 and 20.0% Aloe vera in the culture medium maintained the levels of mRNA for SOD, CAT and PRDX6 in follicles similar to the control group. In conclusion, Aloe vera maintains the morphology of secondary follicles cultured in vitro for 18 days, and 2.5% of Aloe vera promotes antrum formation, increases follicular viability and maintains mRNA expression for SOD and PRDX6 after cultivation.
publishDate 2021
dc.date.accessioned.fl_str_mv 2021-09-02T00:33:42Z
dc.date.available.fl_str_mv 2021-09-02T00:33:42Z
dc.date.issued.fl_str_mv 2021-05-18
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv Azevedo, V.A.N. Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro. 2021. 105f. Dissertação (Mestrado em Biotecnologia) - Programa de Pós-Graduação em Biotecnologia, Campus de Sobral, Universidade Federal do Ceará, Sobral, 2021.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/60208
identifier_str_mv Azevedo, V.A.N. Efeito do extrato de Aloe Vera no crescimento, viabilidade e na expressão gênica de folículos secundários bovinos cultivados in vitro. 2021. 105f. Dissertação (Mestrado em Biotecnologia) - Programa de Pós-Graduação em Biotecnologia, Campus de Sobral, Universidade Federal do Ceará, Sobral, 2021.
url http://www.repositorio.ufc.br/handle/riufc/60208
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
instacron_str UFC
institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
bitstream.url.fl_str_mv http://repositorio.ufc.br/bitstream/riufc/60208/1/2021_dis_vanazevedo.pdf
http://repositorio.ufc.br/bitstream/riufc/60208/2/license.txt
bitstream.checksum.fl_str_mv b881ca0d88c73321a9785279541e63df
8a4605be74aa9ea9d79846c1fba20a33
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
_version_ 1847793278600085504