Influência de um produto probiótico à base de soja sobre a composição da microbiota intestinal de camundongos submetidos à dieta hiperlipídica
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2017-11-30
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Universidade Estadual Paulista (Unesp)
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Objetivo: analisar o efeito de um produto à base de extrato aquoso de soja, fermentado com Enterococcus faecium CRL 183 e Lactobacillus helveticus 416 e com adição de Bifidobacterium longum ATCC 15707, sobre a composição da microbiota fecal, variação de peso corporal e parâmetros inflamatórios de camundongos submetidos à dieta hiperlipídica. Métodos: O protocolo experimental teve duração de 10 semanas sendo utilizados camundongos machos Swiss (Unib: SW) com oito semanas de idade. Os animais foram divididos em quatro grupos (n=10): C (controle negativo) - animais alimentados com dieta padrão para roedores (DP); OB (controle positivo) - animais induzidos à obesidade por dieta hiperlipídica (DHL - 61,01% do valor energético total de lipídios); OBF (fermentado probiótico) - animais induzidos à obesidade por DHL e que receberam o produto fermentado probiótico; OBP (placebo) - animais induzidos à obesidade por DHL e que receberam o produto placebo (sem os cultivos microbianos). A bebida probiótica e o produto placebo foram produzidos semanalmente e a viabilidade das cepas E. faecium CRL 183, L. helveticus 416 e B. longum ATCC 15707 foi determinada imediatamente após o preparo do produto probiótico (T0) e com sete dias de armazenamento à temperatura de refrigeração (5 ºC). Ao longo do protocolo experimental foram monitorados os seguintes parâmetros: ingestão hídrica e de ração (diariamente), variação do peso corporal (semanalmente) e verificação da glicemia de jejum (final do protocolo). A composição da microbiota intestinal foi avaliada por Reação em Cadeia da Polimerase em Tempo Real (qPCR) no início e no final do protocolo. Após a eutanásia dos animais, as deposições de tecido adiposo branco intra-abdominal (gordura retroperitoneal e epididimal) foram removidas para determinação da massa, área e diâmetro dos adipócitos e níveis de citocinas pró e anti-inflamatórias no tecido adiposo (IL-1β, IL-6, IL-10, TGF-β e TNF-α). Resultados: Foi constatado que a população média de células viáveis dos microrganismos probióticos utilizados no processo fermentativo permaneceu acima de 8,0 logUFC.mL-1 durante todo o protocolo experimental. O grupo OBF se destacou por apresentar peso médio inferior ao do grupo OB até a 9a semana do estudo (p<0,05). A análise histológica evidenciou maior área e diâmetro dos adipócitos nos grupos alimentados por DHL, porém dentre estes, o grupo OBF apresentou valores significativamente menores. Embora a glicemia de jejum não tenha apresentado diferença entre os grupos OB, OBF e OBP (p<0,05), os marcadores inflamatórios indicaram aumento significativo de IL-6 e IL-10 nos grupos OBF e OBP quando comparado aos grupos controles negativo e positivo, além disso, houve maior expressão de TGF-β no grupo OBP (p<0,05). A análise da microbiota por qPCR demonstrou que os animais induzidos à obesidade apresentaram aumento na proporção do gênero Lactobacillus spp. (pertencente ao filo Firmicutes), no entanto, a administração do produto probiótico (grupo OBF) contribuiu com o equilíbrio da microbiota intestinal ao manter a proporção de microrganismos do filo Bacteroidetes e promover um aumento na população de Bifidobacterium spp. Conclusão: Nas condições do estudo, a ingestão regular da bebida probiótica foi capaz de reduzir o ganho de peso corporal, o tamanho dos adipócitos e promover uma melhora no perfil imunológico e no equilíbrio da microbiota fecal dos animais, atuando positivamente no controle da obesidade.
Aim: to investigate the effect of an aqueous soy extract beverage, fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus 416, supplemented with Bifidobacterium longum ATCC 15707, on the composition of fecal microbiota, change in body weight and inflammatory parameters of mice submitted to a hyperlipidic diet. Methods: The length of the experimental protocol was 10 weeks, and it was performed using Swiss type male mice (Unib: SW) at eight weeks of age. The animals were divided into four groups (n = 10): C (negative control) - animals fed with a standard diet for rodents; OB (positive control) - animals induced to obesity by a high-fat diet (HFD - 61.01% of the total energetic value of lipids); OBF (beverage fermented with probiotics) - animals induced to obesity by HFD, that also was fed with the probiotic fermented product; OBP (placebo) - animals induced to obesity by HFD , that also was fed with the placebo product (the same product without microbial cultures). The beverage fermented with probiotics and the placebo product were produced weekly and the viability of its microbial strains (E. faecium CRL 183, L. helveticus 416 and B. longum ATCC 15707) was determined immediately after its preparation (T0) and after seven days of storage at controlled temperature (5 °C). During the experimental protocol, the following parameters were monitored: water and food intake (daily), body weight variation (weekly) and fasting glycemia (at the end of the protocol). The composition of the intestinal microbiota was evaluated by Real-Time Polymerase Chain Reaction (qPCR) at the beginning, middle and at the end of the protocol. After the animals’ euthanasia, the deposition of intra-abdominal white adipose tissue (retroperitoneal and epidydimal fat) was removed to determine the mass, area and diameter of adipocytes and levels of pro and anti-inflammatory cytokines in the adipose tissue (IL-1β, IL-6, IL-10, TGF-β and TNF-α). Results: The OBF group showed a lower mean weight than the OB group until the 9th week of the study (p <0.05). The histological analysis evidenced a larger area and diameter of the adipocytes in the groups fed by HFD, however, among these, the OBF group showed significantly lower values. Although fasting glycemia did not differ between OB, OBF and OBP groups (p <0.05), inflammatory markers indicated a significant increase in IL-10 and IL-6 within the OBF and OBP groups when compared to the negative and positive control groups, in addition, there was higher expression of TGF-β in the OBP group (p<0.05). Analysis of fecal microbiota by qPCR showed that animals which were induced to obesity had an increase in the proportion of the genus Lactobacillus spp. (belonging to the Firmicutes phylum), moreover, administration of the beverage fermented with probiotic (OBF group) appeared to contribute to intestinal balance by maintaining the proportion of the Bacteroidetes phylum microorganisms and promoting a subtle increase in the number of bifidobacteria. Conclusion: Under the conditions of the study, the regular intake of the probiotic beverage could reduce body weight gain, the size of adipocytes and promote an improvement in immune profile and balance of the fecal microbiota of animals, acting positively in the control of obesity.
Aim: to investigate the effect of an aqueous soy extract beverage, fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus 416, supplemented with Bifidobacterium longum ATCC 15707, on the composition of fecal microbiota, change in body weight and inflammatory parameters of mice submitted to a hyperlipidic diet. Methods: The length of the experimental protocol was 10 weeks, and it was performed using Swiss type male mice (Unib: SW) at eight weeks of age. The animals were divided into four groups (n = 10): C (negative control) - animals fed with a standard diet for rodents; OB (positive control) - animals induced to obesity by a high-fat diet (HFD - 61.01% of the total energetic value of lipids); OBF (beverage fermented with probiotics) - animals induced to obesity by HFD, that also was fed with the probiotic fermented product; OBP (placebo) - animals induced to obesity by HFD , that also was fed with the placebo product (the same product without microbial cultures). The beverage fermented with probiotics and the placebo product were produced weekly and the viability of its microbial strains (E. faecium CRL 183, L. helveticus 416 and B. longum ATCC 15707) was determined immediately after its preparation (T0) and after seven days of storage at controlled temperature (5 °C). During the experimental protocol, the following parameters were monitored: water and food intake (daily), body weight variation (weekly) and fasting glycemia (at the end of the protocol). The composition of the intestinal microbiota was evaluated by Real-Time Polymerase Chain Reaction (qPCR) at the beginning, middle and at the end of the protocol. After the animals’ euthanasia, the deposition of intra-abdominal white adipose tissue (retroperitoneal and epidydimal fat) was removed to determine the mass, area and diameter of adipocytes and levels of pro and anti-inflammatory cytokines in the adipose tissue (IL-1β, IL-6, IL-10, TGF-β and TNF-α). Results: The OBF group showed a lower mean weight than the OB group until the 9th week of the study (p <0.05). The histological analysis evidenced a larger area and diameter of the adipocytes in the groups fed by HFD, however, among these, the OBF group showed significantly lower values. Although fasting glycemia did not differ between OB, OBF and OBP groups (p <0.05), inflammatory markers indicated a significant increase in IL-10 and IL-6 within the OBF and OBP groups when compared to the negative and positive control groups, in addition, there was higher expression of TGF-β in the OBP group (p<0.05). Analysis of fecal microbiota by qPCR showed that animals which were induced to obesity had an increase in the proportion of the genus Lactobacillus spp. (belonging to the Firmicutes phylum), moreover, administration of the beverage fermented with probiotic (OBF group) appeared to contribute to intestinal balance by maintaining the proportion of the Bacteroidetes phylum microorganisms and promoting a subtle increase in the number of bifidobacteria. Conclusion: Under the conditions of the study, the regular intake of the probiotic beverage could reduce body weight gain, the size of adipocytes and promote an improvement in immune profile and balance of the fecal microbiota of animals, acting positively in the control of obesity.
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Obesidade, Probiótico, Microbiota intestinal, Citocinas inflamatórias