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
Autor(a) principal: | |
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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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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 |
status_str |
publishedVersion |
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) instacron:CUB |
instname_str |
Centro Universitário Braz Cubas (CUB) |
instacron_str |
CUB |
institution |
CUB |
reponame_str |
Repositório do Centro Universitário Braz Cubas |
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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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1798311341538148352 |