Efeitos do kefir sobre a produção de citocinas e óxido nítrico a partir de macrófagos peritoneais de ratos com diabetes mellitus induzido por estreptozotocina
Data
2014-11-30
Autores
Maciel, Fabiane Romano 
Orientadores
Higa, Elisa Mieko Suemitsu
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Diabetes mellitus é um grave problema de saúde global. Estão previstos entre 366 e 439 milhões de casos em 2030, em todo o mundo. A produção excessiva de espécies reativas de oxigênio no diabetes, decorrente da hiperglicemia e do estresse oxidativo induz a peroxidação lipídica e prejudica a resposta imune. O Kefir (K) é um produto lácteo fermentado, considerado um probiótico, que apresenta propriedades imunomoduladoras. Este estudo teve como objetivo investigar o efeito da ingestão de K sobre a produção de citocinas e de óxido nítrico em sobrenadante de cultura de macrófagos peritoneais extraídos de ratos com diabetes mellitus induzido por estreptozotocina (STZ). Para isso, ratos Wistar, machos, adultos receberam única
injeção de STZ (45 mg/kg, por via intravenosa). Após 72 horas, a concentração de glicose foi medida no sangue da veia caudal por um glicosímetro, sendo considerados diabéticos os animais com glicemia ≥ 200 mg/dL. Os animais foram distribuídos em 4 grupos (n= 5-8 cada): controle (CTL), CTLK, diabético (DM) e DMK, recebendo 1,8mL de K (1010 unidades formadoras de colônias - UFC/g de bactérias lácticas e 104 a 107 UFC/g de levedura) ou o seu veículo por dia, via gavagem, preparado de acordo com as instruções do fabricante; começando no 5º dia de diabetes, durante 8 semanas. Pré e pós-tratamento, os animais foram colocados em gaiolas metabólicas para quantificação dos dados metabólicos (ingestão de água, consumo de ração, diurese, glicemia e massa corporal). Após o tratamento, os animais foram sacrificados em câmara de CO2 e exsanguinados; logo em seguida, a cavidade peritoneal foi preparada e os macrófagos residentes foram coletados e cultivados para análise da capacidade fagocítica, citocinas (IL-10, TNFα, IL-17 e IL-1β) e óxido nítrico (NO). Os resultados são apresentados como média ±EP, analisados por one way ANOVA. O grupo DM, quando comparado ao CTL, apresentou aumento na ingestão hídrica, consumo de ração, diurese e glicemia, demonstrando redução no peso. Após tratamento com K, observamos que essa diferença se manteve entre os grupos DM e CTL, contudo, o grupo DMK apresentou níveis inferiores nos parâmetros metabólicos estudados, exceto para massa corporal, a qual estava aumentada, quando comparado ao DM. Em relação à função
dos macrófagos peritoneais, o grupo DMK apresentou melhora na capacidade fagocítica, aumento na concentração de citocinas e de NO em sobrenadante de cultura celular, comparado ao DM. Estes resultados sugerem que o K tem potencial de modular a resposta imune e ativar a resposta de macrófagos peritoneais em animais diabéticos, o que poderia melhorar a imunocompetência de pacientes acometidos pelo diabetes. O efeito hipoglicemiante deste probiótico poderia ser usado como ferramenta no controle da glicemia, podendo reduzir ou retardar o aparecimento das complicações associadas a esta doença.
Diabetes mellitus is a serious global health problem. There are predicted between 366 and 439 million of cases in 2030 worldwide. The excessive production of reactive oxygen species in diabetes is due to hyperglycemia, and the oxidative stress induces lipid peroxidation and impairs the immune response. Kefir (K) is a fermented dairy product, considered a probiotic, which presents immunomodulatory properties. The aim of this study was to investigate the effect of Kefir ingestion on the cytokines and nitric oxide production, in culture supernatant of peritoneal macrophages from streptozotocin-induced diabetic rats. For this purpose, male adult Wistar rats received a single injection of streptozotocin (45 mg/kg, intravenously). After 72 hours, glucose concentration was measured in the tail vein blood using a glucometer; DM was defined as glycemia ≥ 200 mg/dL. The animals were allocated in four groups (n = 5-8 each): Control (CTL); CTLK; DM; DMK. K (1010 CFU/g of dairy bacteria and 104-107 CFU/g of yeast) or its vehicle were given at 1.8 mL/day by gavage, prepared according to the manufacturer's instructions, started on the 5th day of diabetes, during 8 weeks. Pre and post treatment, the animals were placed in metabolic cages for measurement of the metabolic parameters (water intake, food intake, diuresis, glycemia and body weight). After treatment, the animals were sacrificed in CO2 chamber and xsanguinated, and then, the peritoneal cavity was prepared and the resident macrophages were collected and cultured for analysis of phagocytic capacity, cytokines (IL-10, TNF-α, IL-17 and IL-1β) and nitric oxide (NO). Results are presented as mean ± SEM and analyzed by one way ANOVA. The DM group compared to CTL showed increase in water intake, food intake, glycemia and diuresis, demonstrating reduction in body weight and insulin. After treatment with K, we observed that this difference remained between DM and CTL groups; however, the DMK group had increase in these metabolic parameters, except for body weight and insulin, which were increased when compared to DM. In relation to the function of peritoneal macrophages, the DMK group showed improvement in phagocytic capacity, increasing in the concentration of cytokines and NO compared to DM. These results suggest that K has a potential to modulate the immune response and activate peritoneal macrophages in diabetic animals, which could enhance the immunocompetence of patients affected by diabetes mellitus. The hypoglycemic effect of this probiotic could be used as a tool to control glycemia and can reduce or delay the onset of complications associated with this disease.
Diabetes mellitus is a serious global health problem. There are predicted between 366 and 439 million of cases in 2030 worldwide. The excessive production of reactive oxygen species in diabetes is due to hyperglycemia, and the oxidative stress induces lipid peroxidation and impairs the immune response. Kefir (K) is a fermented dairy product, considered a probiotic, which presents immunomodulatory properties. The aim of this study was to investigate the effect of Kefir ingestion on the cytokines and nitric oxide production, in culture supernatant of peritoneal macrophages from streptozotocin-induced diabetic rats. For this purpose, male adult Wistar rats received a single injection of streptozotocin (45 mg/kg, intravenously). After 72 hours, glucose concentration was measured in the tail vein blood using a glucometer; DM was defined as glycemia ≥ 200 mg/dL. The animals were allocated in four groups (n = 5-8 each): Control (CTL); CTLK; DM; DMK. K (1010 CFU/g of dairy bacteria and 104-107 CFU/g of yeast) or its vehicle were given at 1.8 mL/day by gavage, prepared according to the manufacturer's instructions, started on the 5th day of diabetes, during 8 weeks. Pre and post treatment, the animals were placed in metabolic cages for measurement of the metabolic parameters (water intake, food intake, diuresis, glycemia and body weight). After treatment, the animals were sacrificed in CO2 chamber and xsanguinated, and then, the peritoneal cavity was prepared and the resident macrophages were collected and cultured for analysis of phagocytic capacity, cytokines (IL-10, TNF-α, IL-17 and IL-1β) and nitric oxide (NO). Results are presented as mean ± SEM and analyzed by one way ANOVA. The DM group compared to CTL showed increase in water intake, food intake, glycemia and diuresis, demonstrating reduction in body weight and insulin. After treatment with K, we observed that this difference remained between DM and CTL groups; however, the DMK group had increase in these metabolic parameters, except for body weight and insulin, which were increased when compared to DM. In relation to the function of peritoneal macrophages, the DMK group showed improvement in phagocytic capacity, increasing in the concentration of cytokines and NO compared to DM. These results suggest that K has a potential to modulate the immune response and activate peritoneal macrophages in diabetic animals, which could enhance the immunocompetence of patients affected by diabetes mellitus. The hypoglycemic effect of this probiotic could be used as a tool to control glycemia and can reduce or delay the onset of complications associated with this disease.
Descrição
Citação
MACIEL, Fabiane Romano. Efeitos do kefir sobre a produção de citocinas e óxido nítrico a partir de macrófagos peritoneais de ratos com diabetes mellitus induzido por estreptozotocina. 2014. Dissertação (Mestrado) - Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, 2014.