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Master's Dissertation
DOI
https://doi.org/10.11606/D.60.2013.tde-24062013-154134
Document
Author
Full name
Lívia Cristina Hernandes
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2013
Supervisor
Committee
Antunes, Lusania Maria Greggi (President)
Tavares, Denise Crispim
Fonseca, Maria Jose Vieira
Title in Portuguese
Avaliação da citotoxicidade, genotoxicidade, antigenotoxicidade e expressão dos genes iNos e COX-2 em ratos tratados com a polpa do fruto de Solanum sessiliflorum Dunal
Keywords in Portuguese
carotenoides
compostos fenólicos
doxorrubicina
maná-cubiu
mutagênese
Abstract in Portuguese
O cubiuzeiro (Solanum sessiliflorum Dunal) é uma planta nativa da Amazônia, utilizada na medicina popular no tratamento de queimaduras e no controle da glicemia e colesterolemia. Análises fitoquímicas da polpa dos frutos do cubiuzeiro, conhecidos como maná-cubiu, revelaram que este apresenta em sua constituição carotenoides, compostos fenólicos e vitaminas. Devido aos poucos estudos sobre a atividade biológica do fruto e à presença de compostos com propriedades antioxidantes, os objetivos deste trabalho foram avaliar os efeitos da polpa do maná-cubiu sobre a estabilidade genômica, parâmetros de estresse oxidativo, expressão de genes pró-inflamatórios (iNos e COX-2) em ratos Wistar e determinar a presença de elementos químicos na polpa liofilizada por ICP-MS. Os animais receberam, por gavagem, a polpa liofilizada do maná-cubiu (125, 250, 375 ou 500 mg/kg p.c.) durante 14 dias. No último dia de tratamento foi administrada intraperitonealmente salina ou doxorrubicina (DXR, 16 mg/kg p.c.), usada como agente indutor de danos, e após 24 horas os animais foram eutanasiados. O sangue periférico e a medula óssea foram usados no teste do micronúcleo. No ensaio do cometa foram analisados fígado, coração e sangue periférico. Fígado e coração foram utilizados nas análises das substâncias reativas ao ácido tiobarbitúrico (TBARS) e glutationa reduzida (GSH) e na avaliação da expressão de RNAm dos genes COX-2 e iNos. Os resultados mostraram que a polpa do maná-cubiu não foi mutagênica nem citotóxica à medula óssea e ao sangue periférico dos animais. Nas doses 250 e 375 mg/kg p.c., a polpa do maná-cubiu reduziu o número de células micronucleadas induzidas pela DXR no sangue periférico, enquanto que na medula óssea somente a dose de 375 mg/kg p.c. foi antimutagênica. A polpa do maná-cubiu não foi genotóxica no fígado, coração e sangue periférico, e os animais tratados com a associação da polpa do maná-cubiu e DXR apresentaram uma redução de danos ao DNA. Nas doses de 250, 375 e 500 mg/kg p.c., a polpa do maná-cubiu foi capaz de reduzir a peroxidação lipídica induzida pela DXR em células hepáticas, mas não alterou as concentrações de GSH no fígado e no coração. Os dados obtidos da reação em cadeia da polimerase em tempo real referentes ao gene COX-2 mostraram que a polpa do maná-cubiu não modulou a transcrição deste gene. Ainda, a polpa liofilizada do maná-cubiu apresentou em sua composição a presença de elementos-traço como zinco, manganês, selênio, cobre e crômio, mas não em quantidades significativas. Os compostos bioativos presentes na polpa do maná-cubiu, como carotenoides e compostos fenólicos, podem estar relacionados com os efeitos protetores em certos tecidos e doses de maná-cubiu observados neste estudo.
Title in English
Cytotoxicity, genotoxicity and antigenotoxicity evaluations and gene expression of iNos and COX-2 in rats treated with the fruit pulp of Solanum sessiliflorum Dunal
Keywords in English
carotenoids
doxorubicin
maná-cubiu
mutagenesis
phenolic compounds
Abstract in English
Cubiuzeiro (Solanum sessiliflorum Dunal) is a native plant from Amazonian Forest, used in folk medicine to treat burns and to control cholesterol and blood glucose levels. Phytochemical analysis of the cubiuzeiro fruit, known as maná-cubiu, revealed that its composition exhibits carotenoids, phenolic compounds and vitamins. Due to the few studies that have assessed the biological activity and the presence of compounds with antioxidant properties in this fruit, the objectives of this study were to evaluate the effects of maná-cubiu pulp on genomic stability, oxidative stress parameters and expression of pro-inflammatory genes (iNos and COX-2) in Wistar rats and to determine the presence of chemical elements in the lyophilized pulp by ICP-MS. The animals received lyophilized maná-cubiu pulp (125, 250, 375 or 500 mg/kg b.w.) by gavage for 14 days. On the last day of treatment, saline or doxorubicin (DXR, 16 mg/kg b.w.), used as a genotoxic agent, were administered intraperitoneally, and after 24 hours the animals were euthanized. Peripheral blood and bone marrow were used in the micronucleus test. In the comet assay, liver, heart and peripheral blood cells were analyzed. Liver and heart tissues were used to analyze the thiobarbituric acid reactive substances and reduced glutathione (GSH) and to evaluate the mRNA expression of COX-2 and iNos genes. The results showed that maná-cubiu pulp was not mutagenic or cytotoxic in bone marrow and peripheral blood of the animals. At 250 and 375 mg/kg b.w. doses, the maná-cubiu pulp reduced the number of micronucleated cells induced by DXR in peripheral blood, while only the 375 mg/kg b.w. dose was antimutagenic in bone marrow cells. The maná-cubiu pulp was not genotoxic on liver, heart and peripheral blood cells, and the animals treated with maná-cubiu and DXR exhibited lower levels of DNA damage. At 250, 375 and 500 mg/kg b.w. doses, maná-cubiu pulp was able to reduce lipidic peroxidation induced by DXR on liver cells, however did not change the GSH levels on heart and liver cells. The data obtained by real time polymerase chain reaction (RT-qPCR) from COX-2 gene showed that maná-cubiu pulp did not alter the expression of this gene. Also, the maná-cubiu pulp presented trace elements as zinc, manganese, selenium, copper and chromium, but not in significant amounts. Bioactive compounds in the maná-cubiu pulp as carotenoids and phenolic compounds may be associated with the protective effect in certain tissues and doses of maná-cubiu shown in this study.
 
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Dissertacao_Resumida.pdf (683.83 Kbytes)
Publishing Date
2013-07-02
 
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  • HERNANDES, L. C., et al. Antigenotoxic evaluation of the fruit pulp maná-cubiu (Solanun sessiliflorum Dunal) in Wistar rats by the comet assay. In 5th International Symposium of Post Graduation and Research, Ribeirão Preto-SP, 2012. CD do 5th International Symposium of Post Graduation and Research.Ribeirão Preto-SP, 2012. Abstract. Available from: http://www.fcfrp.usp.br.
  • HERNANDES, L. C., et al. Cardioprotective effects of maná-cubiu fruit (Solanum sessiliflorum Dunal) in Wistar rats. In 6th Congress of the International Society of Nutrigenetics/Nutrigenomics, São Paulo-SP, 2012. Journal of Nutrigenetics and Nutrigenomics.New York : Karger, 2012. Abstract.
  • HERNANDES, L. C., et al. Maná-cubiu pulp fruit reduces lipid peroxidation induced by doxorubicin in heart and liver of Wisar rats. In 2nd Ibero-American Meeting on Toxicology and Environmental Health, Ribeirão Preto-SP, 2013. Book of Abstracts of 2nd Ibero-American Meeting on Toxicology and Environmental Health., 2013. Abstract.
  • HERNANDES, L. C., et al. Study of the antigenotoxic and cytotoxic activities of the fruit Solanum sessiliflorum by the micronucleus test in rats. In 51st Annual Meeting Society of Toxicology & ExpoTox, San Francisco-USA, 2012. Program Book of 51st Annual Meeting Society of Toxicology & ExpoTox.San Francisco-USA : Editora da SOT, 2012. Abstract. Available from: http://www.toxicology.org.
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