Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro

Detalhes bibliográficos
Autor(a) principal: Jaques, Jeandre Augusto dos Santos
Data de Publicação: 2010
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/11133
Resumo: The exposure to the cigarette smoke xenobiotics is related with a compromised immune system and the development of many diseases such as Chronic Obstructive Pulmonary Disease (COPD) and lung cancer. The NTPDase and acetylcholinesterase (AChE) are important enzymes involved in the regulation of immune system and alterations in their activities have been showed in many diseases including the related with cigarette smoke-exposure. The curcumin (Cur), polyphenol isolated from the rhizomes of Curcuma longa and widely employed in the culinary and oriental traditional medicine, has many biological activities such as antioxidant, anticarcinogenic and antiinflammatory. Considering the relevance of lymphocytes in the establishment and progression of immune responses, the purpose of this study was to evaluate the effect of curcumin in the NTPDase and AChE activities in peripheral lymphocytes (PL) and lung lymphocytes (LL) from rats exposed to cigarette smoke. The establishment of a protocol to lung lymphocytes separation using the lung tissue was performed with the objective of optimize the cellular viability and the purity of separation, using as references the protocols for separation of human peripheral and lung lymphocytes. The experimental procedure to evaluate the effect of curcumin on the NTPDase and AChE in lymphocytes from rats exposed to the cigarette smoke was divided in two sets of experiments. In the first set, the animals were randomly divided into four groups: Vehicle; Cur 12.5 mg/kg; Cur 25 mg/kg; Cur 50 mg/kg. In the second experimental set, the animals were divided into 5 groups: Vehicle; smoke exposed; smoke and Cur 12.5 mg/kg; smoke and Cur 25 mg/kg; smoke and Cur 50 mg/kg. Curcumin was diluted with corn oil, administered by oral gavage, not exceeding 1 ml/kg body weight. The treatment with curcumin and cigarette smoke was carried out once a day, 5 days each week, during 30 days. It was administered the curcumin or corn oil, and approximately 10 minutes later, the smoking groups were exposed to the sidestream smoke of four commercial cigarettes (nicotine 0.9 mg, tar 10 mg each) inside a whole-body smoke exposure chamber. Control animals were placed in an equal chamber for the same amount of time. After thirty days, the animals were euthanized, the lung was removed and the peripheral blood collected for separation of lymphocytes. With the standardization of the protocol to the separation of lung lymphocytes it was achieved a percentage of lymphocytes higher than 82% and it was observed a strong correlation between the enzyme activity using ATP and ADP as substrates, which indicates that the results can be expressed in milligram of protein, millions of cells and millions of viable cells. The results from the experiments with the cigarette smoke exposure demonstrated that the treatment with curcumin prevented the alterations observed in the cigarette smoke-exposed animals such as the decrease of ATP and ADP hydrolysis in PL and LL, and the increase of AChE activity in PL. We suggest that the treatment with curcumin was protective, since the high concentrations of ATP are positively related with inflammation and tissue damage, and the emphysematous damages observed in cigarette smoke-exposed was not observed in the groups treated with curcumin.
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spelling 2017-04-242017-04-242010-08-19JAQUES, Jeandre Augusto dos Santos. Effect of curcumin in the ntpdase and acetylcholinesterase activity in cigarette smoke exposed rats. 2010. 120 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2010.http://repositorio.ufsm.br/handle/1/11133The exposure to the cigarette smoke xenobiotics is related with a compromised immune system and the development of many diseases such as Chronic Obstructive Pulmonary Disease (COPD) and lung cancer. The NTPDase and acetylcholinesterase (AChE) are important enzymes involved in the regulation of immune system and alterations in their activities have been showed in many diseases including the related with cigarette smoke-exposure. The curcumin (Cur), polyphenol isolated from the rhizomes of Curcuma longa and widely employed in the culinary and oriental traditional medicine, has many biological activities such as antioxidant, anticarcinogenic and antiinflammatory. Considering the relevance of lymphocytes in the establishment and progression of immune responses, the purpose of this study was to evaluate the effect of curcumin in the NTPDase and AChE activities in peripheral lymphocytes (PL) and lung lymphocytes (LL) from rats exposed to cigarette smoke. The establishment of a protocol to lung lymphocytes separation using the lung tissue was performed with the objective of optimize the cellular viability and the purity of separation, using as references the protocols for separation of human peripheral and lung lymphocytes. The experimental procedure to evaluate the effect of curcumin on the NTPDase and AChE in lymphocytes from rats exposed to the cigarette smoke was divided in two sets of experiments. In the first set, the animals were randomly divided into four groups: Vehicle; Cur 12.5 mg/kg; Cur 25 mg/kg; Cur 50 mg/kg. In the second experimental set, the animals were divided into 5 groups: Vehicle; smoke exposed; smoke and Cur 12.5 mg/kg; smoke and Cur 25 mg/kg; smoke and Cur 50 mg/kg. Curcumin was diluted with corn oil, administered by oral gavage, not exceeding 1 ml/kg body weight. The treatment with curcumin and cigarette smoke was carried out once a day, 5 days each week, during 30 days. It was administered the curcumin or corn oil, and approximately 10 minutes later, the smoking groups were exposed to the sidestream smoke of four commercial cigarettes (nicotine 0.9 mg, tar 10 mg each) inside a whole-body smoke exposure chamber. Control animals were placed in an equal chamber for the same amount of time. After thirty days, the animals were euthanized, the lung was removed and the peripheral blood collected for separation of lymphocytes. With the standardization of the protocol to the separation of lung lymphocytes it was achieved a percentage of lymphocytes higher than 82% and it was observed a strong correlation between the enzyme activity using ATP and ADP as substrates, which indicates that the results can be expressed in milligram of protein, millions of cells and millions of viable cells. The results from the experiments with the cigarette smoke exposure demonstrated that the treatment with curcumin prevented the alterations observed in the cigarette smoke-exposed animals such as the decrease of ATP and ADP hydrolysis in PL and LL, and the increase of AChE activity in PL. We suggest that the treatment with curcumin was protective, since the high concentrations of ATP are positively related with inflammation and tissue damage, and the emphysematous damages observed in cigarette smoke-exposed was not observed in the groups treated with curcumin.A exposição aos xenobióticos da fumaça do cigarro está relacionada com o comprometimento do sistema imune e o desenvolvimento de diversas doenças como a Doença Pulmonar Obstrutiva Crônica (DPOC) e câncer de pulmão. A NTPDase e a acetilcolinesterase (AChE) são importantes enzimas envolvidas na regulação do sistema imune e alterações nas suas atividades tem sido descritas em várias doenças incluindo as relacionadas à exposição à fumaça do cigarro. A curcumina (Cur), polifenol obtido a partir de rizomas de Curcuma longa e amplamente utilizado na culinária e na medicina tradicional oriental, possui muitas atividades biológicas como antioxidante, anticarcinogênica e antiinflamatória. Tendo em vista a relevância dos linfócitos no estabelecimento e progressão de respostas imunes, o objetivo deste estudo foi avaliar o efeito da curcumina na atividade das enzimas NTPDase e AChE em linfócitos periféricos (PL) e linfócitos pulmonares (LL) de ratos expostos à fumaça do cigarro. A padronização da separação dos linfócitos a partir de tecido pulmonar foi realizada com o objetivo de otimizar a viabilidade celular e a pureza da separação, utilizando-se como base os protocolos de separação de linfócitos periféricos e pulmonares de seres humanos. Os procedimentos experimentais para a avaliação do efeito da curcumina sob a atividade das enzimas NTPDase e AChE em linfócitos de ratos expostos à fumaça do cigarro foram divididos em duas etapas. Primeiramente, os animais foram divididos aleatoriamente em quatro grupos: veículo; Cur 12.5 mg/kg, Cur 25 mg/kg; Cur 50 mg/kg. Na segunda etapa experimental os animais foram divididos aleatoriamente em cinco grupos: veículo; cigarro; cigarro e Cur 12.5 mg/kg; cigarro e Cur 25 mg/kg; cigarro e Cur 50 mg/kg. A curcumina foi diluída em óleo de milho, administrada por gavagem oral, não excedendo 1 ml/kg de massa corpórea. O tratamento com a curcumina e com a fumaça do cigarro foi realizada uma vez por dia, cinco dias por semana, durante trinta dias. A curcumina foi administrada e, após aproximadamente dez minutos, os grupos fumantes eram expostos à fumaça de quatro cigarros comerciais (0,9 mg nicotina, 10 mg alcatrão cada) dentro de uma câmara de exposição. Os animais não expostos à fumaça do cigarro foram colocados em uma câmara equivalente, pelo mesmo período. Após trinta dias, os animais foram eutanasiados, os pulmões removidos e o sangue periférico coletado para a separação dos linfócitos. Com a padronização da técnica para a separação de linfócitos pulmonares foi obtido um percentual de linfócitos superior a 82% e ainda observou-se uma forte correlação entre a atividade da enzima usando ATP e ADP como substrato o que indica que os resultados podem ser expressos tanto em miligramas de proteína, quanto em milhões de células quanto em milhões de células viáveis. Os resultados dos experimentos com a fumaça do cigarro demonstraram que o tratamento com a curcumina preveniu as alterações observadas nos animais expostos à fumaça do cigarro como a diminuição na hidrólise de ATP e ADP em PL e LL, e o aumento da atividade da AChE em PL. Analisando os resultados obtidos, sugere-se que o tratamento com a curcumina foi protetor, uma vez que as altas concentrações de ATP estão positivamente relacionadas com inflamação e dano tecidual, e as lesões enfisematosas observadas no grupo exposto à fumaça do cigarro não foram observadas com os tratamentos com a curcumina.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em Ciências Biológicas: Bioquímica ToxicológicaUFSMBRBioquímicaLinfócitosCurcuminaNTPDaseAcetilcolinesteraseFumaça de cigarroLymphocytesCurcuminNTPDaseAcetylcholinesteraseCigarette smokeCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICAEfeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarroEffect of curcumin in the ntpdase and acetylcholinesterase activity in cigarette smoke exposed ratsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisLeal, Daniela Bitencourt Rosahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706487E7Garcia, Solange Cristinahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790220Y7Lopes, Sonia Teresinha dos Anjoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4794864D7http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4257075A6Jaques, Jeandre Augusto dos Santos200800000002400500500500500ebd6d0bc-4e14-43fd-8d4e-ad9ebe640af2b9cef9e5-7151-48ba-b5b4-a90397b33c2908641410-3b67-4735-9a1f-8563f344fcd6aac588d9-5a32-46a9-b6f9-2f297a059022info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALJAQUES, JEANDRE AUGUSTO DOS SANTOS.pdfapplication/pdf1753237http://repositorio.ufsm.br/bitstream/1/11133/1/JAQUES%2c%20JEANDRE%20AUGUSTO%20DOS%20SANTOS.pdf525ee8206532b3f94a66721c0b013c42MD51TEXTJAQUES, JEANDRE AUGUSTO DOS SANTOS.pdf.txtJAQUES, JEANDRE AUGUSTO DOS SANTOS.pdf.txtExtracted texttext/plain170661http://repositorio.ufsm.br/bitstream/1/11133/2/JAQUES%2c%20JEANDRE%20AUGUSTO%20DOS%20SANTOS.pdf.txt4296690f22f7dac332e9844d7fb66c8cMD52THUMBNAILJAQUES, JEANDRE AUGUSTO DOS SANTOS.pdf.jpgJAQUES, JEANDRE AUGUSTO DOS SANTOS.pdf.jpgIM Thumbnailimage/jpeg5331http://repositorio.ufsm.br/bitstream/1/11133/3/JAQUES%2c%20JEANDRE%20AUGUSTO%20DOS%20SANTOS.pdf.jpg33dcf7efe239c26a55d845dd18b773c8MD531/111332022-07-04 12:25:53.266oai:repositorio.ufsm.br:1/11133Repositório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132022-07-04T15:25:53Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.por.fl_str_mv Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro
dc.title.alternative.eng.fl_str_mv Effect of curcumin in the ntpdase and acetylcholinesterase activity in cigarette smoke exposed rats
title Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro
spellingShingle Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro
Jaques, Jeandre Augusto dos Santos
Linfócitos
Curcumina
NTPDase
Acetilcolinesterase
Fumaça de cigarro
Lymphocytes
Curcumin
NTPDase
Acetylcholinesterase
Cigarette smoke
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
title_short Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro
title_full Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro
title_fullStr Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro
title_full_unstemmed Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro
title_sort Efeito da curcumina sob a atividade das enzimas ntpdase e acetilcolinesterase em linfócitos de ratos expostos à fumaça do cigarro
author Jaques, Jeandre Augusto dos Santos
author_facet Jaques, Jeandre Augusto dos Santos
author_role author
dc.contributor.advisor1.fl_str_mv Leal, Daniela Bitencourt Rosa
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706487E7
dc.contributor.referee1.fl_str_mv Garcia, Solange Cristina
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790220Y7
dc.contributor.referee2.fl_str_mv Lopes, Sonia Teresinha dos Anjos
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4794864D7
dc.contributor.authorLattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4257075A6
dc.contributor.author.fl_str_mv Jaques, Jeandre Augusto dos Santos
contributor_str_mv Leal, Daniela Bitencourt Rosa
Garcia, Solange Cristina
Lopes, Sonia Teresinha dos Anjos
dc.subject.por.fl_str_mv Linfócitos
Curcumina
NTPDase
Acetilcolinesterase
Fumaça de cigarro
topic Linfócitos
Curcumina
NTPDase
Acetilcolinesterase
Fumaça de cigarro
Lymphocytes
Curcumin
NTPDase
Acetylcholinesterase
Cigarette smoke
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
dc.subject.eng.fl_str_mv Lymphocytes
Curcumin
NTPDase
Acetylcholinesterase
Cigarette smoke
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
description The exposure to the cigarette smoke xenobiotics is related with a compromised immune system and the development of many diseases such as Chronic Obstructive Pulmonary Disease (COPD) and lung cancer. The NTPDase and acetylcholinesterase (AChE) are important enzymes involved in the regulation of immune system and alterations in their activities have been showed in many diseases including the related with cigarette smoke-exposure. The curcumin (Cur), polyphenol isolated from the rhizomes of Curcuma longa and widely employed in the culinary and oriental traditional medicine, has many biological activities such as antioxidant, anticarcinogenic and antiinflammatory. Considering the relevance of lymphocytes in the establishment and progression of immune responses, the purpose of this study was to evaluate the effect of curcumin in the NTPDase and AChE activities in peripheral lymphocytes (PL) and lung lymphocytes (LL) from rats exposed to cigarette smoke. The establishment of a protocol to lung lymphocytes separation using the lung tissue was performed with the objective of optimize the cellular viability and the purity of separation, using as references the protocols for separation of human peripheral and lung lymphocytes. The experimental procedure to evaluate the effect of curcumin on the NTPDase and AChE in lymphocytes from rats exposed to the cigarette smoke was divided in two sets of experiments. In the first set, the animals were randomly divided into four groups: Vehicle; Cur 12.5 mg/kg; Cur 25 mg/kg; Cur 50 mg/kg. In the second experimental set, the animals were divided into 5 groups: Vehicle; smoke exposed; smoke and Cur 12.5 mg/kg; smoke and Cur 25 mg/kg; smoke and Cur 50 mg/kg. Curcumin was diluted with corn oil, administered by oral gavage, not exceeding 1 ml/kg body weight. The treatment with curcumin and cigarette smoke was carried out once a day, 5 days each week, during 30 days. It was administered the curcumin or corn oil, and approximately 10 minutes later, the smoking groups were exposed to the sidestream smoke of four commercial cigarettes (nicotine 0.9 mg, tar 10 mg each) inside a whole-body smoke exposure chamber. Control animals were placed in an equal chamber for the same amount of time. After thirty days, the animals were euthanized, the lung was removed and the peripheral blood collected for separation of lymphocytes. With the standardization of the protocol to the separation of lung lymphocytes it was achieved a percentage of lymphocytes higher than 82% and it was observed a strong correlation between the enzyme activity using ATP and ADP as substrates, which indicates that the results can be expressed in milligram of protein, millions of cells and millions of viable cells. The results from the experiments with the cigarette smoke exposure demonstrated that the treatment with curcumin prevented the alterations observed in the cigarette smoke-exposed animals such as the decrease of ATP and ADP hydrolysis in PL and LL, and the increase of AChE activity in PL. We suggest that the treatment with curcumin was protective, since the high concentrations of ATP are positively related with inflammation and tissue damage, and the emphysematous damages observed in cigarette smoke-exposed was not observed in the groups treated with curcumin.
publishDate 2010
dc.date.issued.fl_str_mv 2010-08-19
dc.date.accessioned.fl_str_mv 2017-04-24
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dc.identifier.citation.fl_str_mv JAQUES, Jeandre Augusto dos Santos. Effect of curcumin in the ntpdase and acetylcholinesterase activity in cigarette smoke exposed rats. 2010. 120 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2010.
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/11133
identifier_str_mv JAQUES, Jeandre Augusto dos Santos. Effect of curcumin in the ntpdase and acetylcholinesterase activity in cigarette smoke exposed rats. 2010. 120 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2010.
url http://repositorio.ufsm.br/handle/1/11133
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