Avaliação da atividade anti-inflamatória intestinal da dieta enriquecida com Hibiscus esculentus L. no modelo de inflamação intestinal induzida por TNBS em ratos
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Data
2018-02-27
Autores
Oliva, Késsien Regina Sander
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Universidade Estadual Paulista (Unesp)
Resumo
A Doença Inflamatória Intestinal engloba duas principais doenças sendo elas a Doença de Crohn e a Retocolite Ulcerativa onde ambas se caracterizam por uma inflamação crônica do intestino, com períodos de exacerbação seguidos de intervalos prologados com remissão dos sintomas. Sua etiologia é considerada multifatorial e ainda pouco elucidada. Considerando-se que não existe cura, que os fármacos utilizados são de alto custo, além de apresentarem sérios efeitos colaterais e ainda muitos pacientes não responderem a esses tratamentos disponíveis, a busca por estratégias complementares de prevenção e tratamento dessa doença que combinem eficácia e segurança, como o uso de alimentos funcionais, compostos antioxidantes e compostos bioativos se apresentam como uma perspectiva promissora. Com base nisso, a espécie vegetal Hibiscus esculentus L. conhecida popularmente como quiabo, foi selecionada para o presente estudo com o objetivo de avaliar a atividade anti-inflamatória intestinal das dietas enriquecidas com o fruto no modelo experimental da inflamação intestinal induzida por ácido trinitrobenzenosulfônico em ratos. Para tanto, a farinha dos frutos foi incorporada na dieta de ratos Wistar machos nas concentrações de 5% e de 10% por 28 dias antes e 7 dias após a indução do processo inflamatório intestinal. Os animais foram mortos no 35º dia e seus cólons foram extraídos. Para a caracterização da atividade anti-inflamatória intestinal foram analisados parâmetros macroscópicos (escore, extensão da lesão, relação peso/comprimento do cólon, ocorrência de diarreia e aderência do cólon a órgãos adjacentes), análises microscópicas (hematoxilina-eosina e PAS/Alcian Blue para avaliação de muco) e parâmetros bioquímicos (atividade das enzimas mieloperoxidase e fosfatase alcalina e quantificação da glutationa total). Complementarmente foi realizada a análise fitoquímica do extrato etanólico 70% dos frutos de Hibiscus esculentus, a atividade sequestradora de radicais livres utilizando o modelo do 2,2-difenil-1-picril-hidrazila (DPPH), e a avaliação da atividade antioxidante pelo ensaio de peroxidação lipídica em membranas de cérebros de ratos. As avaliações bioquímicas mostraram que a dieta nas duas concentrações foi capaz de evitar a depleção dos níveis de glutationa, mas não diminuiu a atividade das enzimas mieloperoxidase e fosfatase alcalina em nenhuma das concentrações testadas. A dieta nas duas concentrações foi capaz de evitar depleção de muco e foi observada recuperação tecidual pelas análises microscópicas. A dieta na concentração de 10% diminuiu a relação peso/comprimento cólico e extensão da lesão. As análises fitoquímicas mostram a presença de fenóis, taninos, flavonóis, flavononas, xantonas, terpenóides. No ensaio de DPPH a farinha apresentou uma IC50 de 302.68ug/ml vs. 1.89ug/ml do ácido gálico, enquanto que no modelo de peroxidação lipídica, uma IC50 de 196,8μg/ml, onde no mesmo teste, a quercetina, flavonoide antioxidante usado como referência, apresentou IC50 de 1,02μg/ml. Esses resultados sugerem que a suplementação dietética com os frutos de Hibiscus esculentus nas concentrações de 5% e de 10% na ração foi responsável pela melhora do processo inflamatório cólico induzido por ácido trinitrobenzenosulfônico em ratos. Esses efeitos estão associados às propriedades antioxidantes da espécie que podem estar relacionadas com a presença de compostos fenólicos, sugerindo que essa espécie é um alimento funcional promissor para o tratamento complementar da doença inflamatória intestinal, propondo a necessidade de que mais estudos sejam realizados para confirmarem e esclarecerem esses efeitos.
Inflammatory Bowel Disease comprises two major diseases, Crohn´s disease and ulcerative colitis where both are characterized by chronic inflammation of the intestine, with periods of exacerbation followed by prolonged intervals with remission of symptoms. This disease is multifactorial with complex and not fully elucidated etiology. Considering that there is no cure and the drugs may cause serious side effects, and many patients do not respond to the available treatments, researches focused on the development of new treatment strategies are important. The pursuit of complementary strategy of prevention and treatment of disease that combine effectiveness and safety, such as the use of functional foods, antioxidant compounds and bioactive compounds if known as a promising perspective. Based on this, a vegetable species Hibiscus esculentus L., populary known as okra, was selected for this study with the objective to evaluate the activity of enriched diet in the experimental model of intestinal inflammation induced by trinitrobenzenesulfonic acid in rats. For this, a fruit flour was incorporated in the diet of male Wistar rats in concentrations of 5% and 10% for 28 days before and 7 days after of induction of intestinal inflammation by TNBS. The animals were killed on the 35th day and their colons were removed. For the characterization of the intestinal antiinflammatory activity, were performed macroscopic studies (score, lesion length, weight ratio length of the colon, occurrence of diarrhea and grip), microscopic analysis (hematoxylin-eosin and PAS/Alcian Blue for evaluation of mucus) and biochemical parameters (myeloperoxidase and alkaline phosphatase enzyme and quantification of total glutathione). Complementarily, it was made a photochemical analysis of the 70% ethanolic extract of Hibiscus esculentus fruits, a free radical scavenging activity using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) model and an evaluation of antioxidant activity by the lipid peroxidation assay in membranes of rat brains. The biochemical evaluations showed that the diet at both concentrations was able to prevent depletion of glutathione but did not decrease the activity of the enzymes myeloperoxidase and alkaline phosphatase in any of the concentrations. The diet at both concentrations was able to avoid mucus depletion and tissue recovery was observed by microscopic analysis. The 10% concentration diet decreased the colic weight/length ratio and extension of the lesion. The phytochemical analyzes showed a presence of phenols, tannins, flavonols, flavonones, xanthones, terpenoids, DPPH showed an IC50 of 302.68ug/ml vs. 1.89ug /ml of gallic acid and a model of lipid peroxidation, the extract showed an IC50 of 196.8μg/ml, where in the same test, the quercetin, antioxidant flavonoid used as reference, showed an IC50 of 1.02μg/ml. These results suggest that dietary supplementation with fruits flour of Hibiscus esculentus at concentrations of 5% and 10% in feed was responsible for the improvement of the colonic inflammatory process induced by trinitrobenzenesulfonic acid in rats. These results are associated as antioxidant properties of the specie that may be related to a presence of phenolic, suggesting that the specie is a functional food for the complementary treatment of inflammatory bowel disease, suggesting the need for further studies to confirm and clarify the effects.
Inflammatory Bowel Disease comprises two major diseases, Crohn´s disease and ulcerative colitis where both are characterized by chronic inflammation of the intestine, with periods of exacerbation followed by prolonged intervals with remission of symptoms. This disease is multifactorial with complex and not fully elucidated etiology. Considering that there is no cure and the drugs may cause serious side effects, and many patients do not respond to the available treatments, researches focused on the development of new treatment strategies are important. The pursuit of complementary strategy of prevention and treatment of disease that combine effectiveness and safety, such as the use of functional foods, antioxidant compounds and bioactive compounds if known as a promising perspective. Based on this, a vegetable species Hibiscus esculentus L., populary known as okra, was selected for this study with the objective to evaluate the activity of enriched diet in the experimental model of intestinal inflammation induced by trinitrobenzenesulfonic acid in rats. For this, a fruit flour was incorporated in the diet of male Wistar rats in concentrations of 5% and 10% for 28 days before and 7 days after of induction of intestinal inflammation by TNBS. The animals were killed on the 35th day and their colons were removed. For the characterization of the intestinal antiinflammatory activity, were performed macroscopic studies (score, lesion length, weight ratio length of the colon, occurrence of diarrhea and grip), microscopic analysis (hematoxylin-eosin and PAS/Alcian Blue for evaluation of mucus) and biochemical parameters (myeloperoxidase and alkaline phosphatase enzyme and quantification of total glutathione). Complementarily, it was made a photochemical analysis of the 70% ethanolic extract of Hibiscus esculentus fruits, a free radical scavenging activity using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) model and an evaluation of antioxidant activity by the lipid peroxidation assay in membranes of rat brains. The biochemical evaluations showed that the diet at both concentrations was able to prevent depletion of glutathione but did not decrease the activity of the enzymes myeloperoxidase and alkaline phosphatase in any of the concentrations. The diet at both concentrations was able to avoid mucus depletion and tissue recovery was observed by microscopic analysis. The 10% concentration diet decreased the colic weight/length ratio and extension of the lesion. The phytochemical analyzes showed a presence of phenols, tannins, flavonols, flavonones, xanthones, terpenoids, DPPH showed an IC50 of 302.68ug/ml vs. 1.89ug /ml of gallic acid and a model of lipid peroxidation, the extract showed an IC50 of 196.8μg/ml, where in the same test, the quercetin, antioxidant flavonoid used as reference, showed an IC50 of 1.02μg/ml. These results suggest that dietary supplementation with fruits flour of Hibiscus esculentus at concentrations of 5% and 10% in feed was responsible for the improvement of the colonic inflammatory process induced by trinitrobenzenesulfonic acid in rats. These results are associated as antioxidant properties of the specie that may be related to a presence of phenolic, suggesting that the specie is a functional food for the complementary treatment of inflammatory bowel disease, suggesting the need for further studies to confirm and clarify the effects.
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Palavras-chave
TNBS, Hibiscus esculentus, quiabo, alimentos funcionais, doença Inflamatória Intestinal, inflammatory bowel disease, TNBS, Hibiscus esculentus, okra, functional foods