Lesão aterosclerótica, capacidade antioxidante e histopatologia de camundongos apoe -/- alimentados com açaí (Euterpe Edulis Martius) e submetidos ao treinamento físico
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Data
2011-03-17
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Universidade Federal de Viçosa
Resumo
A doença arterial aterosclerótica é a maior causa de morte no mundo. A prática de exercício físico e o consumo de alimentos ricos em antioxidantes estão associados a um decréscimo na incidência de eventos cardiovasculares. O objetivo deste trabalho foi avaliar os efeitos de uma dieta a base dos frutos de açaí (Euterpe edulis) associada ao treinamento físico, no processo de aterosclerose, na capacidade antioxidante do figado e na histopatologia do fígado e rim em camundongos adultos C57BL/6 knockout para o gene da Apolipoproteína E(apoE-/-). Foram utilizados 28 camundongos (ApoE-/-) e 10 camundongos C57Bl/6, com 21 semanas de idade, submetidos a 12 semanas de corrida em esteira (atingindo ao final 60 minutos/sessão/dia, 5 dias/semana e velocidade de 15m/min) e dieta com 2% de açaí. Antes de iniciar o tratamento alguns animais de ambas as linhagens (T1 - 4 camundongos ApoE - / - e T2 - 5 C57Bl/6) sofreram eutanásia para avaliação do nível inicial da lesão aterosclerótica. Após este procedimento o restante dos animais foram divididos em 5 grupos: C- Controle negativo animais C57Bl/6, sem 2% de açaí, não-treinado (n=4); C+ Controle positivo animais ApoE -/- sem 2% de açaí, não-treinado (n=4 ); G1 animais ApoE -/- com 2% de açaí, não-treinado (n=6); G2 animais ApoE -/- com 2% de açaí, treinados (n=6); G3 animais ApoE -/- sem 2% de açaí, treinados (n=6). Os frutos do açaí foram obtidos no entorno do Parque Estadual da Serra do Brigadeiro (PESB), localizada na região da zona da mata do estado de Minas Gerais. A colheita, pós-colheita e despolpamento dos frutos foram realizados conforme orientações da Embrapa para os frutos do Euterpe oleracea, açaí da Amazônia. Após todas as fases, a polpa do fruto do Euterpe. edulis foi embalada e liofilizada para utilização na dieta. Na polpa liofilizada foram determinadas também a composição centesimal e uma triagem fitoquímica para as principais classes de metabólitos secundários: flavonóides, cumarinas, taninos, antraquinonas, óleos essenciais, triterpenos/esteróides, saponinas e alcalóides. O peso e o consumo alimentar dos camundongos foram monitorados semanalmente. Ao final do tratamento os animais sofreram eutanásia, o sangue foi coletado por punção na aorta abdominal e imediatamente centrifugado para obtenção do soro. A artéria aorta foi removida, desde a porção torácica até a abdominal, para análise de lesões
aterosclerótica. O fígado, rins, coração e aorta foram removidos e lavados em
solução fisiológica e em seguida, conservados em paraformaldeido 10% para
posteriores análises histopatológicas. Um fragmento do fígado foi congelado
em nitrogênio líquido e mantido a -80ºC até a análise de atividade enzimática
da catalase e superoxido dismutase. O fruto liofilizado apresentou 8,45 % de
umidade, 5,28% proteína, 49,35% de lipídeos e 42,86% de carboidratos. A triagem fitoquímica do extrato de E. edulis foi positiva para a classe de metabólitos como os flavonóides, polifenóis e saponinas, e negativo para alcalóides, esteróides/triterpenos, cumarinas e antraquinonas. Não houve diferença estatística entre os grupos para peso corporal, consumo de dieta e peso do fígado. Considerando o perfil lipídico plasmático, não houve diferença entre os grupos quanto aos valores de colesterol total, triglicerídeos e HDL em relação ao grupo controle positivo (C+). A porcentagem de área lesionada foi menor apenas no grupo G3, comparado com o C+ (p<0,05). A atividade da catalase não foi diferente entre os grupos, mas foi observada menor atividade da SOD nos dois grupos que fizeram atividade física em comparação ao C+(p<0,05). Os grupos treinados apresentaram menor porcentagem de esteatose no fígado em relação ao C+. No rim, os tratamentos não provocaram alterações no número e área dos glomérulos. Este trabalho sugere que a quantidade de açaí proposta na dieta dos animais não alterou positivamente o processo de aterosclerose neste modelo animal. No entanto, o programa de corrida em esteira foi capaz de provocar redução na área das lesões ateroscleróticas e na atividade da SOD, reduzindo também a deposição de gordura no tecido hepático.
Atherosclerotic arterial disease is the leading cause of death worldwide. The practice of physical exercise and consumption of foods rich in antioxidants are associated with a decrease in the incidence of cardiovascular events. The purpose of this study was to evaluate the effects of a diet based on the açaí fruit (Euterpe edulis) associated with physical exercise on the process of atherosclerosis, on the antioxidant capacity of the liver, and on the liver and kidney histopathology of adult mice C57BL/6 knockout for the Apolipoprotein E gene (apoE-/-).Twenty eight mice (ApoE-/ -) and 10 C57BL/6 with 21 weeks of age were used and submitted to 12 weeks of moderate exercise (treadmill running reaching 60 minutes/session/day during 5 days/week and speed of 15m/min) and diet with 2% acai. Before starting the treatment some animals of both strains (T1 - 4 mice ApoE - / - and T2 - 5 C57BL / 6) were sacrificed to assess the initial level of atherosclerotic lesions. After this procedure the remaining animals were divided into five groups: C-Negative control animals C57BL/6 without 2% açai, untrained (n = 4); C + Positive control - animals ApoE -/- without 2% açai, non-trained (n = 4); G1 animals ApoE -/-, 2% of acai, untrained (n = 6); G2 animals ApoE -/-, 2% of acai trained (n = 6 ) and G3 animals ApoE -/-, 2% without açai, trained (n = 6). The açaí fruit were obtained in the vicinity of the Parque Estadual da Serra do Brigadeiro (PESB), located in the Zona da Mata of Minas Gerais. Harvesting, post-harvest selection of fruit and pulping were performed according to guidelines from Embrapa for Euterpe oleracea fruit, Amazon Açaí. After these stages, the pulp of the fruit of Euterpe. edulis was packed and freeze-dried for use in the diet. The freeze-dried pulp was used to determine its chemical composition and to proceed a phytochemical screening to identify the main classes of secondary metabolites such as flavonoids, coumarins, tannins, anthraquinones, essential oils, triterpenes/steroids, saponins and alkaloids. The weight and food intake of the mice were evaluated weekly. At the end of treatment the animals were euthanized, blood was collected by puncturing the abdominal aorta and immediately centrifuged to obtain the serum. The aorta was removed from the chest to the abdominal portion for the analysis of atherosclerotic lesions. The liver, kidneys, heart and aorta were removed and washed in saline solution, then preserved in 10% paraformaldehyde for further histopathological examinations. A fragment of the liver was frozen in liquid nitrogen and kept at - 80 °C until analysis of the catalase and superoxide dismutase enzymatic activities. The dried fruit had 8.45% moisture, 5.28% protein, 49.35% fat and 42.86% carbohydrates. The phytochemical screening of the extract of E. edulis was positive for the class of metabolites such as flavonoids, polyphenols and saponins, and negative for alkaloids, steroids / triterpenes, coumarins and anthraquinones. There were no statistical differences between the groups for the body and liver weights and for the food intake. Considering the plasma lipid profile, there were no differences between groups in relation to total cholesterol, triglycerides and HDL compared to positive control group (C+). The percentage of injured area was lower only in G3 compared with C + (p <0.05). The catalase activity was not different between groups, but it was observed a lower SOD activity in both physical activity groups (p <0.05). The trained groups showed a lower percentage of steatosis than C+ group. In the kidney the treatments did not significantly change the number and area of the glomeruli. This study suggests that the quantity of acaí fruit added in the animal´s diet did not altered positively the atherosclerotic process in this animal model. However, the treadmill running program was able to promote reductions in the area of atherosclerotic lesions and in SOD activity, also altering fat deposition in liver tissue.
Atherosclerotic arterial disease is the leading cause of death worldwide. The practice of physical exercise and consumption of foods rich in antioxidants are associated with a decrease in the incidence of cardiovascular events. The purpose of this study was to evaluate the effects of a diet based on the açaí fruit (Euterpe edulis) associated with physical exercise on the process of atherosclerosis, on the antioxidant capacity of the liver, and on the liver and kidney histopathology of adult mice C57BL/6 knockout for the Apolipoprotein E gene (apoE-/-).Twenty eight mice (ApoE-/ -) and 10 C57BL/6 with 21 weeks of age were used and submitted to 12 weeks of moderate exercise (treadmill running reaching 60 minutes/session/day during 5 days/week and speed of 15m/min) and diet with 2% acai. Before starting the treatment some animals of both strains (T1 - 4 mice ApoE - / - and T2 - 5 C57BL / 6) were sacrificed to assess the initial level of atherosclerotic lesions. After this procedure the remaining animals were divided into five groups: C-Negative control animals C57BL/6 without 2% açai, untrained (n = 4); C + Positive control - animals ApoE -/- without 2% açai, non-trained (n = 4); G1 animals ApoE -/-, 2% of acai, untrained (n = 6); G2 animals ApoE -/-, 2% of acai trained (n = 6 ) and G3 animals ApoE -/-, 2% without açai, trained (n = 6). The açaí fruit were obtained in the vicinity of the Parque Estadual da Serra do Brigadeiro (PESB), located in the Zona da Mata of Minas Gerais. Harvesting, post-harvest selection of fruit and pulping were performed according to guidelines from Embrapa for Euterpe oleracea fruit, Amazon Açaí. After these stages, the pulp of the fruit of Euterpe. edulis was packed and freeze-dried for use in the diet. The freeze-dried pulp was used to determine its chemical composition and to proceed a phytochemical screening to identify the main classes of secondary metabolites such as flavonoids, coumarins, tannins, anthraquinones, essential oils, triterpenes/steroids, saponins and alkaloids. The weight and food intake of the mice were evaluated weekly. At the end of treatment the animals were euthanized, blood was collected by puncturing the abdominal aorta and immediately centrifuged to obtain the serum. The aorta was removed from the chest to the abdominal portion for the analysis of atherosclerotic lesions. The liver, kidneys, heart and aorta were removed and washed in saline solution, then preserved in 10% paraformaldehyde for further histopathological examinations. A fragment of the liver was frozen in liquid nitrogen and kept at - 80 °C until analysis of the catalase and superoxide dismutase enzymatic activities. The dried fruit had 8.45% moisture, 5.28% protein, 49.35% fat and 42.86% carbohydrates. The phytochemical screening of the extract of E. edulis was positive for the class of metabolites such as flavonoids, polyphenols and saponins, and negative for alkaloids, steroids / triterpenes, coumarins and anthraquinones. There were no statistical differences between the groups for the body and liver weights and for the food intake. Considering the plasma lipid profile, there were no differences between groups in relation to total cholesterol, triglycerides and HDL compared to positive control group (C+). The percentage of injured area was lower only in G3 compared with C + (p <0.05). The catalase activity was not different between groups, but it was observed a lower SOD activity in both physical activity groups (p <0.05). The trained groups showed a lower percentage of steatosis than C+ group. In the kidney the treatments did not significantly change the number and area of the glomeruli. This study suggests that the quantity of acaí fruit added in the animal´s diet did not altered positively the atherosclerotic process in this animal model. However, the treadmill running program was able to promote reductions in the area of atherosclerotic lesions and in SOD activity, also altering fat deposition in liver tissue.
Descrição
Palavras-chave
Aterosclerose, Antioxidantes, Açaí, Exercício físico, Atherosclerosis, Antioxidants, Açaí, Physical exercise
Citação
CASTRO, Cynthia Aparecida de. Atherosclerotic lesions, histopathology and antioxidant capacity of mice ApoE -/- fed with açaí (Euterpe Edulis Martius) and submitted to physical training. 2011. 84 f. Dissertação (Mestrado em Aspectos sócio-culturais do movimento humano; Aspectos biodinâmicos do movimento humano) - Universidade Federal de Viçosa, Viçosa, 2011.