Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da USP |
Texto Completo: | http://www.teses.usp.br/teses/disponiveis/25/25149/tde-25112019-222647/ |
Resumo: | The fluoride ion (F), when ingested in excess, may alter various cellular functions. As liver is an organ of high metabolic activity, it has become the target of investigations on the side effects of F, related to changes in the expression of metabolic proteins, increase in oxidative stress and changes in mitochondrial function. Thus, the objective of the present work was to evaluate the protein expression profile of liver mitochondria from rats chronically exposed to two F concentrations through the drinking water (15 or 50 mgF/L) for 20 or 60 days. Thirty-six liver samples from 21-day-old male Wistar rats were divided into 6 groups (n= 6 animals per group) according to the concentration of F administered in drinking water (0, 15 or 50 mg/L) and the treatment time (20 or 60 days). Mitochondria were extracted from hepatic tissue and prepared for quantitative label-free proteomic analysis (Xevo Q-TOF, Waters). PLGS software was used to detect changes in protein expression among the different groups. Most of the changes were observed in the metabolic pathways such as glycolysis, gluconeogenesis, - oxidation, urea cycle, tricarboxylic acids cycle and electron transport chain, in addition alterations in proteins involved in the antioxidant system, calcium homeostasis and apoptosis. The dose of 15 mgF/L, when administered for 20 days, reduced glycolysis, which was counterbalanced by an increase in other energetic pathways. At 60 days, however, an increase in all energy pathways was observed. On the other hand, the dose of 50 mgF/L, when administered for 20 days, reduced the enzymes involved in all energetic pathways, indicating a lower rate of energy production, with less generation of ROS and consequent reduction of antioxidant enzymes. However, when the 50 mgF/L dose was administered for 60 days, an increase in energy metabolism was seen but in general no changes were observed in the antioxidant enzymes. Except for the group treated with 50 mgF/L for 20 days, all the other groups had alterations in proteins in attempt to maintain calcium homeostasis and avoid apoptosis. Thus, the results suggest that the organism seems to adapt to the effects of F over time, activating pathways to reduce the toxicity of this ion. Ultimately, our findings corroborate the safety of the use of fluoride for caries control. |
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Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periodsAnálise proteômica das mitocôndrias de fígados de ratos expostos à exposição crônica por fluoreto em dois períodos experimentaisAntioxidant systemEnergy metabolismFígadoFluoretoFluorideLiverMetabolismo energéticoMitochondriaMitocôndriaSistema antioxidanteThe fluoride ion (F), when ingested in excess, may alter various cellular functions. As liver is an organ of high metabolic activity, it has become the target of investigations on the side effects of F, related to changes in the expression of metabolic proteins, increase in oxidative stress and changes in mitochondrial function. Thus, the objective of the present work was to evaluate the protein expression profile of liver mitochondria from rats chronically exposed to two F concentrations through the drinking water (15 or 50 mgF/L) for 20 or 60 days. Thirty-six liver samples from 21-day-old male Wistar rats were divided into 6 groups (n= 6 animals per group) according to the concentration of F administered in drinking water (0, 15 or 50 mg/L) and the treatment time (20 or 60 days). Mitochondria were extracted from hepatic tissue and prepared for quantitative label-free proteomic analysis (Xevo Q-TOF, Waters). PLGS software was used to detect changes in protein expression among the different groups. Most of the changes were observed in the metabolic pathways such as glycolysis, gluconeogenesis, - oxidation, urea cycle, tricarboxylic acids cycle and electron transport chain, in addition alterations in proteins involved in the antioxidant system, calcium homeostasis and apoptosis. The dose of 15 mgF/L, when administered for 20 days, reduced glycolysis, which was counterbalanced by an increase in other energetic pathways. At 60 days, however, an increase in all energy pathways was observed. On the other hand, the dose of 50 mgF/L, when administered for 20 days, reduced the enzymes involved in all energetic pathways, indicating a lower rate of energy production, with less generation of ROS and consequent reduction of antioxidant enzymes. However, when the 50 mgF/L dose was administered for 60 days, an increase in energy metabolism was seen but in general no changes were observed in the antioxidant enzymes. Except for the group treated with 50 mgF/L for 20 days, all the other groups had alterations in proteins in attempt to maintain calcium homeostasis and avoid apoptosis. Thus, the results suggest that the organism seems to adapt to the effects of F over time, activating pathways to reduce the toxicity of this ion. Ultimately, our findings corroborate the safety of the use of fluoride for caries control.O íon fluoreto (F), quando ingerido em excesso, pode alterar várias funções celulares. Como o fígado é um órgão de alta atividade metabólica, tornou-se alvo de investigações sobre os efeitos colaterais do F, relacionados a alterações na expressão de proteínas metabólicas, aumento do estresse oxidativo e alterações na função mitocondrial. Assim, o objetivo do presente trabalho foi avaliar o perfil de expressão proteica de mitocôndrias hepáticas de ratos expostos cronicamente a duas concentrações de F na água de beber (15 ou 50 mgF / L) por 20 ou 60 dias. Trinta e seis amostras de fígado de ratos Wistar machos com 21 dias de idade foram divididos em 6 grupos (n= 6 animais por grupo) de acordo com a concentração de F administrada em água potável (0, 15 ou 50 mg / L) e o tratamento tempo (20 ou 60 dias). As mitocôndrias foram extraídas do tecido hepático e preparadas para análise proteômica quantitativa sem marcadores (Xevo Q-TOF, Waters). O software PLGS foi utilizado para detectar mudanças na expressão de proteínas entre os diferentes grupos. A maioria das alterações foi observada nas vias metabólicas como glicólise, gliconeogênese, -oxidação, ciclo da ureia, ciclo dos ácidos tricarboxílicos e cadeia de transporte de elétrons, além de alterações nas proteínas envolvidas no sistema antioxidante e homeostase do cálcio. A dose de 15 mgF/L, quando administrada por 20 dias, reduziu a glicólise, que foi contrabalançada pelo aumento de outras vias energéticas. Aos 60 dias, no entanto, um aumento em todas as vias de energia foi observado. Por outro lado, a dose de 50 mgF/L, quando administrada por 20 dias, reduziu as enzimas envolvidas em todas as vias energéticas, indicando menor taxa de produção de energia, com menor geração de ROS e consequente redução de enzimas antioxidantes. No entanto, quando a dose de 50 mgF/L foi administrada por 60 dias, observou-se um aumento no metabolismo energético, mas, em geral, não foram observadas alterações nas enzimas antioxidantes. Com exceção do grupo tratado com 50 mgF/L por 20 dias, todos os demais grupos apresentaram alterações nas proteínas na tentativa de manter a homeostase do cálcio e evitar a apoptose. Assim, os resultados sugerem que o organismo parece se adaptar aos efeitos do F ao longo do tempo, ativando vias para reduzir a toxicidade desse íon. Em última análise, nossos achados corroboram com a segurança do uso do flúor no controle da cárie.Biblioteca Digitais de Teses e Dissertações da USPBuzalaf, Marilia Afonso RabeloAraújo, Tamara Teodoro2019-04-04info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://www.teses.usp.br/teses/disponiveis/25/25149/tde-25112019-222647/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2024-08-02T13:27:02Zoai:teses.usp.br:tde-25112019-222647Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212024-08-02T13:27:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods Análise proteômica das mitocôndrias de fígados de ratos expostos à exposição crônica por fluoreto em dois períodos experimentais |
title |
Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods |
spellingShingle |
Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods Araújo, Tamara Teodoro Antioxidant system Energy metabolism Fígado Fluoreto Fluoride Liver Metabolismo energético Mitochondria Mitocôndria Sistema antioxidante |
title_short |
Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods |
title_full |
Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods |
title_fullStr |
Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods |
title_full_unstemmed |
Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods |
title_sort |
Proteomic analysis of mitochondria from rat liver exposed to chronic fluoride exposure in two experimental periods |
author |
Araújo, Tamara Teodoro |
author_facet |
Araújo, Tamara Teodoro |
author_role |
author |
dc.contributor.none.fl_str_mv |
Buzalaf, Marilia Afonso Rabelo |
dc.contributor.author.fl_str_mv |
Araújo, Tamara Teodoro |
dc.subject.por.fl_str_mv |
Antioxidant system Energy metabolism Fígado Fluoreto Fluoride Liver Metabolismo energético Mitochondria Mitocôndria Sistema antioxidante |
topic |
Antioxidant system Energy metabolism Fígado Fluoreto Fluoride Liver Metabolismo energético Mitochondria Mitocôndria Sistema antioxidante |
description |
The fluoride ion (F), when ingested in excess, may alter various cellular functions. As liver is an organ of high metabolic activity, it has become the target of investigations on the side effects of F, related to changes in the expression of metabolic proteins, increase in oxidative stress and changes in mitochondrial function. Thus, the objective of the present work was to evaluate the protein expression profile of liver mitochondria from rats chronically exposed to two F concentrations through the drinking water (15 or 50 mgF/L) for 20 or 60 days. Thirty-six liver samples from 21-day-old male Wistar rats were divided into 6 groups (n= 6 animals per group) according to the concentration of F administered in drinking water (0, 15 or 50 mg/L) and the treatment time (20 or 60 days). Mitochondria were extracted from hepatic tissue and prepared for quantitative label-free proteomic analysis (Xevo Q-TOF, Waters). PLGS software was used to detect changes in protein expression among the different groups. Most of the changes were observed in the metabolic pathways such as glycolysis, gluconeogenesis, - oxidation, urea cycle, tricarboxylic acids cycle and electron transport chain, in addition alterations in proteins involved in the antioxidant system, calcium homeostasis and apoptosis. The dose of 15 mgF/L, when administered for 20 days, reduced glycolysis, which was counterbalanced by an increase in other energetic pathways. At 60 days, however, an increase in all energy pathways was observed. On the other hand, the dose of 50 mgF/L, when administered for 20 days, reduced the enzymes involved in all energetic pathways, indicating a lower rate of energy production, with less generation of ROS and consequent reduction of antioxidant enzymes. However, when the 50 mgF/L dose was administered for 60 days, an increase in energy metabolism was seen but in general no changes were observed in the antioxidant enzymes. Except for the group treated with 50 mgF/L for 20 days, all the other groups had alterations in proteins in attempt to maintain calcium homeostasis and avoid apoptosis. Thus, the results suggest that the organism seems to adapt to the effects of F over time, activating pathways to reduce the toxicity of this ion. Ultimately, our findings corroborate the safety of the use of fluoride for caries control. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-04-04 |
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 |
http://www.teses.usp.br/teses/disponiveis/25/25149/tde-25112019-222647/ |
url |
http://www.teses.usp.br/teses/disponiveis/25/25149/tde-25112019-222647/ |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
|
dc.rights.driver.fl_str_mv |
Liberar o conteúdo para acesso público. info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Liberar o conteúdo para acesso público. |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.coverage.none.fl_str_mv |
|
dc.publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da USP instname:Universidade de São Paulo (USP) instacron:USP |
instname_str |
Universidade de São Paulo (USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Biblioteca Digital de Teses e Dissertações da USP |
collection |
Biblioteca Digital de Teses e Dissertações da USP |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP) |
repository.mail.fl_str_mv |
virginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.br |
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1809091082694164480 |