Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae

Detalhes bibliográficos
Autor(a) principal: Belache, Fernanda de Noronha Sertori
Data de Publicação: 2018
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório do Centro Universitário Braz Cubas
Texto Completo: https://repositorio.cruzeirodosul.edu.br/handle/123456789/2655
Resumo: Alzheimer's disease (AD) is the leading age-related neurodegenerative disease. AD is characterized by neurofibrillary formation arising from Tau protein hyperphosphorylation and the formation of senile plaques by a β-amyloid protein tumor. The model of neurodegeneration with intracerebroventricular injection (icv) of streptozotocin (STZ) is used to study sporadic AD (AED). Drug intoxication is not free, as a consequence, oxidative stress and neuroinflammation. Plant sources with antioxidant properties have been prominent in works related to the development of alternative therapies for a disease. Phenolic compounds present in some species and biological activities antioxidant and antiinflammatory, and may help in reducing brain damage and cognitive impairment observed in AED. The objective of the present study was to evaluate the effect of the crude extract of the fruits of Vaccinium myrtillus L. on the expression of genes that perform functions in the neurogenesis, oxidative and neuroinflammatory processes in an animal model of neurodegeneration induced by the icv administration of STZ. Twenty male Wistar rats were divided into four experimental groups, with the same number of animals in each group (n = 5): group 1, control group, in which the animals received citrate buffer via icv and oral distilled water; group 2 who received STZ injection by icv and oral distilled water; group 3, group that received STZ by icv and were treated with the crude extract of the V. myrtillus fruit orally at the concentration of 200 mg/kg; and group 4 composed of animals that received STZ by icv and were treated with the oral extract at a concentration of 400 mg/kg. The experiment was conducted for 30 days. Behavioral tests were then performed for two months. After these tests, the euthanasia of the animals and the removal of the encephalon were made for hippocampal dissection and RNA extraction. Total tissue RNA was extracted with the Trizol™ Reagent extraction kit in combination with the PureLink® RNA Mini Kit (Life Technologies), converted into complementary DNA (cDNA) by the reverse transcription reaction. Samples were submitted to quantitative polymerase chain reaction (RT-qPCR) for relative quantification of brain-derived neurotrophic factor (BDNF), complement system C3, glutathione peroxidase (GPx) and catalase (CAT) genes. Analyzes showed that there was no significant difference in the expression of BDNF, GPx and CAT genes between the animals that received STZ icv and the control group after two months of treatment with crude extract. There was a significant difference in expression of C3 in the groups that received icv administration of STZ. This result demonstrates the existence of an inflammatory process associated with STZ injection. However, no difference in expression was observed between the groups that received STZ administration and were submitted to treatment with aqueous extract of V. myrtillus fruit, showing that the extract does not influence the regulation of the expression of this gene.
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spelling Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceaeBiotecnologia industrialEstreptozocinaDoença de AlzheimerAntioxidantesEstresse oxidativoCNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERALAlzheimer's disease (AD) is the leading age-related neurodegenerative disease. AD is characterized by neurofibrillary formation arising from Tau protein hyperphosphorylation and the formation of senile plaques by a β-amyloid protein tumor. The model of neurodegeneration with intracerebroventricular injection (icv) of streptozotocin (STZ) is used to study sporadic AD (AED). Drug intoxication is not free, as a consequence, oxidative stress and neuroinflammation. Plant sources with antioxidant properties have been prominent in works related to the development of alternative therapies for a disease. Phenolic compounds present in some species and biological activities antioxidant and antiinflammatory, and may help in reducing brain damage and cognitive impairment observed in AED. The objective of the present study was to evaluate the effect of the crude extract of the fruits of Vaccinium myrtillus L. on the expression of genes that perform functions in the neurogenesis, oxidative and neuroinflammatory processes in an animal model of neurodegeneration induced by the icv administration of STZ. Twenty male Wistar rats were divided into four experimental groups, with the same number of animals in each group (n = 5): group 1, control group, in which the animals received citrate buffer via icv and oral distilled water; group 2 who received STZ injection by icv and oral distilled water; group 3, group that received STZ by icv and were treated with the crude extract of the V. myrtillus fruit orally at the concentration of 200 mg/kg; and group 4 composed of animals that received STZ by icv and were treated with the oral extract at a concentration of 400 mg/kg. The experiment was conducted for 30 days. Behavioral tests were then performed for two months. After these tests, the euthanasia of the animals and the removal of the encephalon were made for hippocampal dissection and RNA extraction. Total tissue RNA was extracted with the Trizol™ Reagent extraction kit in combination with the PureLink® RNA Mini Kit (Life Technologies), converted into complementary DNA (cDNA) by the reverse transcription reaction. Samples were submitted to quantitative polymerase chain reaction (RT-qPCR) for relative quantification of brain-derived neurotrophic factor (BDNF), complement system C3, glutathione peroxidase (GPx) and catalase (CAT) genes. Analyzes showed that there was no significant difference in the expression of BDNF, GPx and CAT genes between the animals that received STZ icv and the control group after two months of treatment with crude extract. There was a significant difference in expression of C3 in the groups that received icv administration of STZ. This result demonstrates the existence of an inflammatory process associated with STZ injection. However, no difference in expression was observed between the groups that received STZ administration and were submitted to treatment with aqueous extract of V. myrtillus fruit, showing that the extract does not influence the regulation of the expression of this gene.A doença de Alzheimer (DA) é a principal doença neurodegenerativa relacionada à idade. A DA é caracterizada pela formação de emaranhados neurofibrilares decorrente da hiperfosforilação da proteína Tau e a formação de placas senis por acúmulo da proteína βamiloide. O modelo de neurodegeneração baseado na injeção intracerebroventricular (icv) de estreptozotocina (STZ) é utilizado para estudar a DA esporádica (DAE). A injeção dessa droga gera radicais livres no encéfalo, e, como consequência, leva a estresse oxidativo e neuroinflamação. Fontes vegetais com propriedades antioxidantes têm se destacado em estudos relacionados ao desenvolvimento de terapias alternativas para a doença. Compostos fenólicos presentes em alguns vegetais e frutos possuem atividade antioxidante e antiinflamatória, podendo auxiliar na diminuição de danos cerebrais e dos prejuízos cognitivos observados na DAE. O objetivo da presente pesquisa foi avaliar o efeito do extrato bruto dos frutos de Vaccinium myrtillus L. sobre a expressão de genes que desempenham funções na neurogênese, processo oxidativo e neuroinflamatório em um modelo animal de neurodegeneração, induzida pela administração icv de STZ. Vinte ratos machos da linhagem Wistar foram divididos em quatro grupos experimentais, com número de animais iguais em cada grupo (n=5): grupo 1, grupo controle, em que os animais receberam tampão citrato por via icv e água destilada via oral; grupo 2 que recebeu injeção icv de STZ (3mg/Kg) e água destilada via oral; grupo 3, grupo que recebeu STZ por injeção icv e os animais foram tratados com o extrato bruto do fruto de V. myrtillus via oral na concentração de 200 mg/kg; e grupo 4 composto por animais que receberam STZ por injeção icv e foram tratados com o extrato via oral na concentração de 400 mg/kg. Os animais foram tratados com extrato durante 30 dias. Após dois meses do fim do tratamento, foi feita a eutanásia dos animais e a remoção do encéfalo para dissecação do hipocampo e extração do RNA. O RNA total dos tecidos foi extraído com o kit de extração Trizol™ Reagent em combinação com PureLink® RNA Mini Kit (Life Technologies) e convertido em DNA complementar (cDNA) por meio da reação de transcrição reversa. As amostras foram submetidas à reação em cadeia da polimerase quantitativa (RT-qPCR) para quantificação relativa da expressão dos genes fator neurotrófico derivado do encéfalo (BDNF), C3 do sistema complemento, glutationa peroxidase (GPx) e catalase (CAT). Análises mostraram que não houve diferença significativa da expressão dos genes BDNF, GPx e CAT entre os animais que receberam STZ icv e o grupo controle após dois meses do tratamento com extrato bruto. Houve diferença significativa de expressão de C3, nos grupos que receberam a administração icv de STZ. Esse resultado demonstra a existência de um processo inflamatório associado a injeção de STZ. Entretanto, não foi verificada diferença de expressão entre os grupos que receberam a administração de STZ e foram submetidos ao tratamento com extrato aquoso do fruto de V. myrtillus mostrando que o extrato não exerce influência na regulação da expressão desse gene.Universidade PositivoBrasilPós-GraduaçãoPrograma de Pós-Graduação em Biotecnologia IndustrialUPPincerati, Márcia Reginahttp://lattes.cnpq.br/1632493068360047Silva, Ilton Santos dahttp://lattes.cnpq.br/2457028554249650Belache, Fernanda de Noronha Sertori2021-08-19T20:28:37Z20182021-08-19T20:28:37Z2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://repositorio.cruzeirodosul.edu.br/handle/123456789/2655porinfo:eu-repo/semantics/openAccessreponame:Repositório do Centro Universitário Braz Cubasinstname:Centro Universitário Braz Cubas (CUB)instacron:CUB2021-08-24T18:37:35Zoai:repositorio.cruzeirodosul.edu.br:123456789/2655Repositório InstitucionalPUBhttps://repositorio.brazcubas.edu.br/oai/requestbibli@brazcubas.edu.bropendoar:2021-08-24T18:37:35Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB)false
dc.title.none.fl_str_mv Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae
title Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae
spellingShingle Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae
Belache, Fernanda de Noronha Sertori
Biotecnologia industrial
Estreptozocina
Doença de Alzheimer
Antioxidantes
Estresse oxidativo
CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
title_short Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae
title_full Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae
title_fullStr Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae
title_full_unstemmed Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae
title_sort Expressão dos genes BDNF, C3, GPx e CAT em um modelo animal de neurodegeneração após tratamento com extrato bruto do fruto de Vaccinium myrtillus L., ericaceae
author Belache, Fernanda de Noronha Sertori
author_facet Belache, Fernanda de Noronha Sertori
author_role author
dc.contributor.none.fl_str_mv Pincerati, Márcia Regina
http://lattes.cnpq.br/1632493068360047
Silva, Ilton Santos da
http://lattes.cnpq.br/2457028554249650
dc.contributor.author.fl_str_mv Belache, Fernanda de Noronha Sertori
dc.subject.por.fl_str_mv Biotecnologia industrial
Estreptozocina
Doença de Alzheimer
Antioxidantes
Estresse oxidativo
CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
topic Biotecnologia industrial
Estreptozocina
Doença de Alzheimer
Antioxidantes
Estresse oxidativo
CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
description Alzheimer's disease (AD) is the leading age-related neurodegenerative disease. AD is characterized by neurofibrillary formation arising from Tau protein hyperphosphorylation and the formation of senile plaques by a β-amyloid protein tumor. The model of neurodegeneration with intracerebroventricular injection (icv) of streptozotocin (STZ) is used to study sporadic AD (AED). Drug intoxication is not free, as a consequence, oxidative stress and neuroinflammation. Plant sources with antioxidant properties have been prominent in works related to the development of alternative therapies for a disease. Phenolic compounds present in some species and biological activities antioxidant and antiinflammatory, and may help in reducing brain damage and cognitive impairment observed in AED. The objective of the present study was to evaluate the effect of the crude extract of the fruits of Vaccinium myrtillus L. on the expression of genes that perform functions in the neurogenesis, oxidative and neuroinflammatory processes in an animal model of neurodegeneration induced by the icv administration of STZ. Twenty male Wistar rats were divided into four experimental groups, with the same number of animals in each group (n = 5): group 1, control group, in which the animals received citrate buffer via icv and oral distilled water; group 2 who received STZ injection by icv and oral distilled water; group 3, group that received STZ by icv and were treated with the crude extract of the V. myrtillus fruit orally at the concentration of 200 mg/kg; and group 4 composed of animals that received STZ by icv and were treated with the oral extract at a concentration of 400 mg/kg. The experiment was conducted for 30 days. Behavioral tests were then performed for two months. After these tests, the euthanasia of the animals and the removal of the encephalon were made for hippocampal dissection and RNA extraction. Total tissue RNA was extracted with the Trizol™ Reagent extraction kit in combination with the PureLink® RNA Mini Kit (Life Technologies), converted into complementary DNA (cDNA) by the reverse transcription reaction. Samples were submitted to quantitative polymerase chain reaction (RT-qPCR) for relative quantification of brain-derived neurotrophic factor (BDNF), complement system C3, glutathione peroxidase (GPx) and catalase (CAT) genes. Analyzes showed that there was no significant difference in the expression of BDNF, GPx and CAT genes between the animals that received STZ icv and the control group after two months of treatment with crude extract. There was a significant difference in expression of C3 in the groups that received icv administration of STZ. This result demonstrates the existence of an inflammatory process associated with STZ injection. However, no difference in expression was observed between the groups that received STZ administration and were submitted to treatment with aqueous extract of V. myrtillus fruit, showing that the extract does not influence the regulation of the expression of this gene.
publishDate 2018
dc.date.none.fl_str_mv 2018
2018
2021-08-19T20:28:37Z
2021-08-19T20:28:37Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
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dc.identifier.uri.fl_str_mv https://repositorio.cruzeirodosul.edu.br/handle/123456789/2655
url https://repositorio.cruzeirodosul.edu.br/handle/123456789/2655
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.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Positivo
Brasil
Pós-Graduação
Programa de Pós-Graduação em Biotecnologia Industrial
UP
publisher.none.fl_str_mv Universidade Positivo
Brasil
Pós-Graduação
Programa de Pós-Graduação em Biotecnologia Industrial
UP
dc.source.none.fl_str_mv reponame:Repositório do Centro Universitário Braz Cubas
instname:Centro Universitário Braz Cubas (CUB)
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instname_str Centro Universitário Braz Cubas (CUB)
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collection Repositório do Centro Universitário Braz Cubas
repository.name.fl_str_mv Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB)
repository.mail.fl_str_mv bibli@brazcubas.edu.br
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