Efeito da proteína do soro de leite submetida à radiação UV-C na colite induzida em modelo animal
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Data
2023-02-24
Autores
Montibeller, Maria Jara [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A colite ulcerativa é uma doença inflamatória intestinal crônica e redicivante, que acomete parte do cólon, impactando a qualidade de vida do paciente. Diversas estratégias não farmacológicas têm sido utilizadas para auxiliar na modulação da microbiota intestinal e reduzir os sintomas dessa doença, com destaque para a ingestão de alimentos com potencial anti-inflamatório. Entre as estratégias dietéticas, a ingestão de proteína do soro de leite foi associada à modulação da microbiota e do sistema imune, tendo remissão dos sintomas da colite. Além disso, entre as tecnologias não térmicas que podem ser utilizadas para a produção de soro de leite, a luz ultravioleta pode aumentar a digestibilidade proteica, auxiliando na absorção de diversos aminoácidos essenciais. O objetivo do estudo foi avaliar o efeito do consumo de proteína de soro de leite (WP) submetida ao tratamento de luz na região do ultravioleta de 200-280 nm (UV-C) na colite aguda induzida em camundongos. Para isso os animais foram divididos em quatro grupos (n=10): (H) animais sadios sem consumo de WP, (C) animais com colite e sem consumo de WP, (W) animais com colite e que receberam WP não submetido à UV-C e (WU) animais com colite que receberam WP submetido à UV-C. O protocolo teve duração total de 14 dias, sendo que o WP foi ingerido durante todo o período experimental e a indução da colite pela ingestão de dextran sulfato de sódio ocorreu a partir do sétimo dia. A evolução da colite foi acompanhada pela determinação do índice de atividade da doença (IAD), variação da relação peso e comprimento e análise histopatológica do cólon, determinação da concentração de citocinas pró e anti-inflamatórias, análise da composição da microbiota fecal ao longo do experimento e análises metabolômicas nos rins e plasma após eutanásia. A determinação de metabólitos foi feita por Espectroscopia de Ressonância Magnética Nuclear de Prótons (RMN de 1H) e a determinação da composição da microbiota fecal foi realizada em sequenciador de nova geração (IlluminaMiseq). O nível de 5% de significância (p≤0,05) foi adotado para todas as tomadas de decisão. Considerando a análise do conjunto de sintomas do IAD, o grupo WU resultou em menor progressão da doença. Os diferentes grupos experimentais não apresentaram diferenças significativas em relação às concentrações de IL-10 e TNF-α. Contudo, os níveis das citocinas inflamatórias IL-2 e IL-6 foram superiores no grupo induzido à colite e sem tratamento (p < 0,05). Não houve alteração na diversidade e riqueza da microbiota antes e após indução da colite nos grupos pertencentes às dietas WP (número de observações). Os resultados revelaram que cada um dos suplementos de WP modula a microbiota intestinal durante a inflamação de maneiras diferentes, com destaque para WP tratados com UV-C que apresentaram menor regressão da progressão nos parâmetros clínicos e conservação na abundância relativa dos gêneros. Além disso, a ingestão de WP tratado com UV-C resultou em modulação positiva da MI, com destaque para aumento de Turicibacter spp. e recuperação da família Ruminococcaceae, Lachnospiraceae_UCG_001 e Blautia, o que provavelmente contribuiu para o controle da colite. As análises de metabólitos nos rins não evidenciaram uma sobrecarga das funções renais devido consumo de WP, e foram encontradas alterações em rotas metabólicas, que podem ter sido uma forma de reduzir a progressão da doença. A maior parte dos componentes encontrados nos rins foram estatisticamente diferentes em suas concentrações, quando comparados os grupos controle H e C (p < 0,05). O grupo W não apresentou diferenças estatísticas com grupo saudável em relação à asparagina, alanina, dimetilamina, isoleucina, fenilalanina e valina. Os resultados da análise de metabólitos no plasma sugeriram que este não é um biomarcador ideal para a colite, devido à semelhança entre os grupos que receberam os tratamentos e o grupo controle. Por fim, pode-se concluir que o consumo de W e WU reduziram a progressão da doença, porém com diferentes respostas nas vias metabólicas e na microbiota intestinal, constituindo uma opção de suplemento alimentar para auxiliar no controle dos sintomas da colite.
Ulcerative colitis is a chronic and relapsing inflammatory bowel disease that affects part of the colon, impacting the patient's quality of life. Several nonpharmacological strategies have been used to help modulate the intestinal microbiota and reduce the symptoms of this disease, with emphasis on the intake of foods with anti-inflammatory agent. Among dietary strategies, whey protein intake was associated with modulation of the microbiota and the immune system, with remission of colitis symptoms. Also, among the nonthermal technologies that can be used for the production of whey, ultraviolet light can increase protein digestibility, aiding in the absorption of several essential amino acids. The aim of the study was to evaluate the effect of consumption of whey protein (WP) subjected to light treatment in the ultraviolet region of 200-280 nm (UV-C) on acute colitis induced in mice. In this regard, the animals were divided into four groups (n=10): (H) healthy animals without WP consumption, (C) animals with colitis and without WP consumption, (W) animals with colitis and that received WP not submitted to UV-C and (WU) animals with colitis that received WP subjected to UV-C. The protocol had a total duration of 14 days. The WP was ingested throughout the experimental period and the induction of colitis by ingestion of sodium dextran sulfate occurred from the seventh day. The evolution of the colitis was followed by the determination of the disease activity index (DAI), variation in the weight and length ratio and histopathological analysis of the colon, determination of the concentration of pro and anti-inflammatory cytokines, analysis of the composition of the fecal microbiota throughout the experiment and metabolomic analysis of kidneys and plasma after euthanasia. The determination of metabolites was performed by Proton Nuclear Magnetic Resonance (1H NMR) and the determination of fecal microbiota composition was performed in a new generation sequencer (IlluminaMiseq). The 5% significance level (p≤0.05) was adopted for all tests hypothesis. Considering the analysis of the set of DAI symptoms, the WU group resulted in less disease progression. The different experimental groups did not show significant difference in relation to the concentrations of IL-10 and TNF-α. However, the levels of the inflammatory cytokines IL-2 and IL-6 were higher in the colitisinduced and untreated groups (p < 0.05). There was no change in the diversity and richness of the microbiota before and after colitis induction in the groups belonging to the WP diets (number of observations). The results revealed that each of the WP supplements modulates the gut microbiota during inflammation in different ways, but UV-C-treated WP was more effective, considering less progression in clinical parameters and conservation in the relative abundance of the genera. In addition, the ingestion of WP treated with UV-C resulted in a positive modulation of gut microbiota, with emphasis on the increase of Turicibacter spp. and abundance recovery in the families Ruminococcaceae , Lachnospiraceae_UCG_001 and Blautia, which probably contributed to the control of colitis. Metabolites analysis in the kidneys did not show an overload of renal functions due to WP consumption, and alterations in metabolic pathways were found, which may have been a way to reduce the progression of the disease. Most of the components found in the kidneys were statisticallydifferent in their concentrations when comparing the control groups (p < 0.05). The W group did not show statistical differences with the healthy group in relation to asparagine, alanine, dimethylamine, isoleucine, phenylalanine and valine. Results of plasma metabolite analysis suggested that this is not an ideal biomarker for colitis. Finally, it can be concluded that the consumption of W and WU reduced the progression of the disease, but with different responses in the metabolic pathways and in the intestinal microbiota, being an option for a food supplement to help control the symptoms of colitis.
Ulcerative colitis is a chronic and relapsing inflammatory bowel disease that affects part of the colon, impacting the patient's quality of life. Several nonpharmacological strategies have been used to help modulate the intestinal microbiota and reduce the symptoms of this disease, with emphasis on the intake of foods with anti-inflammatory agent. Among dietary strategies, whey protein intake was associated with modulation of the microbiota and the immune system, with remission of colitis symptoms. Also, among the nonthermal technologies that can be used for the production of whey, ultraviolet light can increase protein digestibility, aiding in the absorption of several essential amino acids. The aim of the study was to evaluate the effect of consumption of whey protein (WP) subjected to light treatment in the ultraviolet region of 200-280 nm (UV-C) on acute colitis induced in mice. In this regard, the animals were divided into four groups (n=10): (H) healthy animals without WP consumption, (C) animals with colitis and without WP consumption, (W) animals with colitis and that received WP not submitted to UV-C and (WU) animals with colitis that received WP subjected to UV-C. The protocol had a total duration of 14 days. The WP was ingested throughout the experimental period and the induction of colitis by ingestion of sodium dextran sulfate occurred from the seventh day. The evolution of the colitis was followed by the determination of the disease activity index (DAI), variation in the weight and length ratio and histopathological analysis of the colon, determination of the concentration of pro and anti-inflammatory cytokines, analysis of the composition of the fecal microbiota throughout the experiment and metabolomic analysis of kidneys and plasma after euthanasia. The determination of metabolites was performed by Proton Nuclear Magnetic Resonance (1H NMR) and the determination of fecal microbiota composition was performed in a new generation sequencer (IlluminaMiseq). The 5% significance level (p≤0.05) was adopted for all tests hypothesis. Considering the analysis of the set of DAI symptoms, the WU group resulted in less disease progression. The different experimental groups did not show significant difference in relation to the concentrations of IL-10 and TNF-α. However, the levels of the inflammatory cytokines IL-2 and IL-6 were higher in the colitisinduced and untreated groups (p < 0.05). There was no change in the diversity and richness of the microbiota before and after colitis induction in the groups belonging to the WP diets (number of observations). The results revealed that each of the WP supplements modulates the gut microbiota during inflammation in different ways, but UV-C-treated WP was more effective, considering less progression in clinical parameters and conservation in the relative abundance of the genera. In addition, the ingestion of WP treated with UV-C resulted in a positive modulation of gut microbiota, with emphasis on the increase of Turicibacter spp. and abundance recovery in the families Ruminococcaceae , Lachnospiraceae_UCG_001 and Blautia, which probably contributed to the control of colitis. Metabolites analysis in the kidneys did not show an overload of renal functions due to WP consumption, and alterations in metabolic pathways were found, which may have been a way to reduce the progression of the disease. Most of the components found in the kidneys were statisticallydifferent in their concentrations when comparing the control groups (p < 0.05). The W group did not show statistical differences with the healthy group in relation to asparagine, alanine, dimethylamine, isoleucine, phenylalanine and valine. Results of plasma metabolite analysis suggested that this is not an ideal biomarker for colitis. Finally, it can be concluded that the consumption of W and WU reduced the progression of the disease, but with different responses in the metabolic pathways and in the intestinal microbiota, being an option for a food supplement to help control the symptoms of colitis.
Descrição
Palavras-chave
Colite ulcerativa, Soro de leite, Metabolômica, Microbiota intestinal, Tratamento por ultravioleta, Whey protein, Gut microbiota, Metabolomics, Ulcerative colitis