Estudio y comparación de la microbiota intestinal en individuos no celiacos frente a pacientes diagnosticados de enfermedad celiaca = Study and comparison of the intestinal microbiota in non celiac individuals and patients with celiac disease

203 p.La enfermedad celiaca (EC) es una enfermedad inflamatoria del intestino delgado causado por una respuesta inapropiada a las proteínas del gluten del trigo y otras proteínas similares del centeno y la cebada en individuos genéticamente susceptibles. Además del gluten, otros factores ambientales podrían jugar también un papel importante en el desarrollo de la enfermedad celiaca. La presencia de una bacteria de morfología bacilar anclada en la parte alta del intestino delgado de niños con EC, que no aparecía en individuos sanos, planteó por primera vez el posible papel de las bacterias en la patogénesis de la EC. En los últimos años, se ha descrito también la existencia de diferencias cuantitativas y cualitativas en la composición de la microbiota fecal y duodenal entre niños con EC e individuos control. Teniendo en cuenta todas estas evidencias científicas, nos planteamos llevar a cabo un estudio comparativo de la microbiota intestinal en individuos no celiacos frente a pacientes diagnosticados con EC. El estudio de la microbiota duodenal mediante colección de clones de una región del ADNr 16S determinó que el tratamiento de la EC mediante la retirada del gluten de la dieta, ocasiona cambios en la composición de la microbiota del intestino delgado de los pacientes adultos enfermos celiacos tratados. Además, se observó también que la edad es uno de los factores que determina las diferencias entre las comunidades de bacterias de los niños y los adultos, tanto entre individuos no celiacos como entre pacientes diagnosticados de EC. Por otro lado, se llevó a cabo un estudio molecular de la microbiota duodenal capaz de crecer en un medio de cultivo con gluten como única fuente de nitrógeno, para ello se cultivaron biopsias procedentes de niños y adultos tanto sanos como enfermos. No se observó un perfil de especies bacterianas asociadas a la EC o a los individuos sanos, pero si se que la microbiota duodenal capaz de crecer en ese medio estaba dominada por especies del género Streptococcus. Por último, se caracterizó la microbiota fecal tanto de individuo sanos como de pacientes enfermos celiacos activos y tratados mediante electroforesis en gradiente desnaturaliante (DGGE) empleando cebadores universales. Los resultados mostraron una microbiota fecal predominante asociada a los individuos sanos, sugiriendo que los pacientes enfermos celiacos podrían presentar cambios en su microbiota fecal como consecuencia de la enfermedad. Para completar el estudio se caracterizó también la microbiota presente en la muestras fecales mediante el empleo de oligonucleótidos específicos de grupo. Los pacientes enfermos celiacos tratados mostraron una reducción en la diversidad de bacterias fecales del grupo Lactobacillus y del grupo Bifidobacterium frente a los individuos celiacos activos y sanos, lo cual podía ser explicado por la ausencia de gluten en la dieta. Sin embargo, no se encontró una relación definida entre las diferentes especies del género Bifidobacterium, Lactobacillus, Bacteroides y Clostridium con la E

Intestinal microbiota modulation in obesity-related non-alcoholic fatty liver disease

[EN] Obesity and associated comorbidities, including non-alcoholic fatty liver disease (NAFLD), are a major concern to public well-being worldwide due to their high prevalence among the population, and its tendency on the rise point to as important threats in the future. Therapeutic approaches for obesity-associated disorders have been circumscribed to lifestyle modifications and pharmacological therapies have demonstrated limited efficacy. Over the last few years, different studies have shown a significant role of intestinal microbiota (IM) on obesity establishment and NAFLD development. Therefore, modulation of IM emerges as a promising therapeutic strategy for obesity-associated diseases. Administration of prebiotic and probiotic compounds, fecal microbiota transplantation (FMT) and exercise protocols have shown a modulatory action over the IM. In this review we provide an overview of current approaches targeting IM which have shown their capacity to counteract NAFLD and metabolic syndrome features in human patients and animal models.SIThis work was supported by grants to JG-G and SS-C from Ministerio de Economía y Competitividad/FEDER (BFU2017- 87960-R) and Junta de Castilla y León (LE063U16 and GRS 1888/A/18). DP was supported by a fellowship from Junta de Castilla y León co-financed by the European Social Fund. EN was supported by Fundación de Investigación Sanitaria of León. MG-M was supported by CIBERehd contracts. CIBERehd is funded by the Instituto de Salud Carlos III, Spain

Intestinal microbiota transplantation to germ-free mice in a in vivo model of nafld associated with a quercetin treatment

15 p.To select mice donors for intestinal microbiota transplantation based on its metabolic phenotype in response to a high fat diet (HFD) and quercetin treatment (Q). Intestinal microbiota. Resumen de un trabajo resultado del proyecto de investigación financiado por la Consejería de Educación de la Junta de Castilla y León (referencia LE063U16)S

Protective effect of quercetin on high-fat diet-induced non-alcoholic fatty liver disease in mice is mediated by modulating intestinal microbiota imbalance and related gut-liver axis activation

60 p.Gut microbiota is involved in obesity, metabolic syndrome and the progression of nonalcoholic fatty liver disease (NAFLD). It has been recently suggested that the flavonoid quercetin may have the ability to modulate the intestinal microbiota composition, suggesting a prebiotic capacity which highlights a great therapeutic potential in NAFLD. The present study aims to investigate benefits of experimental treatment with quercetin on gut microbial balance and related gut-liver axis activation in a nutritional animal model of NAFLD associated to obesity. C57BL/6J mice were challenged with high fat diet (HFD) supplemented or not with quercetin for 16 weeks. HFD induced obesity, metabolic syndrome and the development of hepatic steatosis as main hepatic histological finding. Increased accumulation of intrahepatic lipids was associated with altered gene expression related to lipid metabolism, as a result of deregulation of their major modulators. Quercetin supplementation decreased insulin resistance and NAFLD activity score, by reducing the intrahepatic lipid accumulation through its ability to modulate lipid metabolism gene expression, cytochrome P450 2E1 (CYP2E1)-dependent lipoperoxidation and related lipotoxicity. Microbiota composition was determined via 16S ribosomal RNA Illumina next-generation sequencing. Metagenomic studies revealed HFD-dependent differences at phylum, class and genus levels leading to dysbiosis, characterized by an increase in Firmicutes/Bacteroidetes ratio and in Gram-negative bacteria, and a dramatically increased detection of Helicobacter genus. Dysbiosis was accompanied by endotoxemia, intestinal barrier dysfunction and gut-liver axis alteration and subsequent inflammatory gene overexpression. Dysbiosis-mediated toll-like receptor 4 (TLR-4)-NF-B signaling pathway activation was associated with inflammasome initiation response and reticulum stress pathway induction. Quercetin reverted gut microbiota imbalance and related endotoxemia-mediated TLR-4 pathway induction, with subsequent inhibition of inflammasome response and reticulum stress pathway activation, leading to the blockage of lipid metabolism gene expression deregulation. Our results support the suitability of quercetin as a therapeutic approach for obesity-associated NAFLD via its anti-inflammatory, antioxidant and prebiotic integrative response.Gut microbiota is involved in obesity, metabolic syndrome and the progression of nonalcoholic fatty liver disease (NAFLD). It has been recently suggested that the flavonoid quercetin may have the ability to modulate the intestinal microbiota composition, suggesting a prebiotic capacity which highlights a great therapeutic potential in NAFLD. The present study aims to investigate benefits of experimental treatment with quercetin on gut microbial balance and related gut-liver axis activation in a nutritional animal model of NAFLD associated to obesity. C57BL/6J mice were challenged with high fat diet (HFD) supplemented or not with quercetin for 16 weeks. HFD induced obesity, metabolic syndrome and the development of hepatic steatosis as main hepatic histological finding. Increased accumulation of intrahepatic lipids was associated with altered gene expression related to lipid metabolism, as a result of deregulation of their major modulators. Quercetin supplementation decreased insulin resistance and NAFLD activity score, by reducing the intrahepatic lipid accumulation through its ability to modulate lipid metabolism gene expression, cytochrome P450 2E1 (CYP2E1)-dependent lipoperoxidation and related lipotoxicity. Microbiota composition was determined via 16S ribosomal RNA Illumina next-generation sequencing. Metagenomic studies revealed HFD-dependent differences at phylum, class and genus levels leading to dysbiosis, characterized by an increase in Firmicutes/Bacteroidetes ratio and in Gram-negative bacteria, and a dramatically increased detection of Helicobacter genus. Dysbiosis was accompanied by endotoxemia, intestinal barrier dysfunction and gut-liver axis alteration and subsequent inflammatory gene overexpression. Dysbiosis-mediated toll-like receptor 4 (TLR-4)-NF-B signaling pathway activation was associated with inflammasome initiation response and reticulum stress pathway induction. Quercetin reverted gut microbiota imbalance and related endotoxemia-mediated TLR-4 pathway induction, with subsequent inhibition of inflammasome response and reticulum stress pathway activation, leading to the blockage of lipid metabolism gene expression deregulation. Our results support the suitability of quercetin as a therapeutic approach for obesity-associated NAFLD via its anti-inflammatory, antioxidant and prebiotic integrative respons

Metabolic Phenotype Transfer Through Gut Microbiota Transplantation from HFD-Fed and Quercetin Treated Donors Modulates Obesity-Related NAFLD in Germ-Free Mice

1 p.Gut microbiota is involved in obesity, metabolic syndrome and nonalcoholic fatty liver disease (NAFLD). Strategies to modulate it, including faecal transplantation and administration of prebiotics as quercetin, are actively being examined. The present study aims to investigate benefits of experimental gut microbiota transfer from donors to germ-free mice (GFm) in a high-fat diet (HFD)-based NAFLD model. Resumen de un trabajo resultado del proyecto de investigación financiado por la Consejería de Educación de la Junta de Castilla y León (referencia LE063U16)S

Akkermansia spp. mediates protection from obesity-associated NAFLD development in germ free mice following intestinal microbiota transplantation from high fat diet and quercetin treated donors

1 p.Dysbiosis and gut-liver axis alteration have been pointed as important contributors to obesity and non-alcoholic fatty liver disease (NAFLD) development. Modulation of intestinal microbiota (IM) emerge as a promising therapeutic strategy for obesity-associated NAFLD. This study aims to determine the effect of IM transplantation and quercetin supplementation in a high fat diet (HFD)-based NAFLD model in germ free mice (GFm). Resumen de un trabajo resultado del proyecto de investigación financiado por la Consejería de Educación de la Junta de Castilla y León (referencia LE063U16)S

Intestinal Microbiota Transplantation From HFD-fed and Quercetin Treated Donors Results in a Complex Metabolic Phenotype Transfer that Modulates Obesity-Related NAFLD in Germ Free Mice

2 p.Intestinal microbiota imbalance and related gut-liver axis activation have been identified as key mechanisms in nonalcoholic fatty liver disease (NAFLD) development. Modulation of intestinal microbiota, through administration of prebiotics or faecal microbiota transplantation, is a promising therapeutic approach for obesity associated diseases including NAFLD. The aim of the present study is to evaluate the benefits of gut microbiota transplantation from donors to germ free mice (GFm) following an experimental treatment with the flavonoid quercetin in a high fat diet (HFD)-based NAFLD model. Resumen de un trabajo resultado del proyecto de investigación financiado por la Consejería de Educación de la Junta de Castilla y León (referencia LE063U16)S

Effect of exercise on gut microbiota and metabolic status modulation in an in vivo model of early obesity and NAFLD

1 p.Childhood obesity is one of the most serious public health concerns from this century, associated with metabolic syndrome, nonalcoholic fatty liver disease (NAFLD) and gut microbiota alterations. Physical exercise improves obesity and NAFLD progression, modulating the gut microbial balance. We aim to investigate the effect of physical exercise on gut microbiota and the metabolic status of an in vivo model of early obesity, metabolic syndrome, and NAFLD. Resumen de un trabajo resultado del proyecto de investigación financiado por la Consejería de Educación de la Junta de Castilla y León (referencia LE063U16)S

Functional Interactions between Gut Microbiota Transplantation, Quercetin, and High‐Fat Diet Determine Non‐Alcoholic Fatty Liver Disease Development in Germ‐Free Mice

[EN] Scope: Modulation of intestinal microbiota has emerged as a new therapeutic approach for non-alcoholic fatty liver disease (NAFLD). Herein, it is addressed whether gut microbiota modulation by quercetin and intestinal microbiota transplantation can influence NAFLD development. Methods and results: Gut microbiota donor mice are selected according to their response to high-fat diet (HFD) and quercetin in terms of obesity and NAFLD-related biomarkers. Germ-free recipients displayed metabolic phenotypic differences derived from interactions between microbiota transplanted, diets, and quercetin. Based on the evaluation of hallmark characteristics of NAFLD, it is found that gut microbiota transplantation from the HFD-non-responder donor and the HFD-fed donor with the highest response to quercetin results in a protective phenotype against HFD-induced NAFLD, in a mechanism that involves gut–liver axis alteration blockage in these receivers. Gut microbiota from the HFD-responder donor predisposed transplanted germ-free mice to NAFLD. Divergent protective and deleterious metabolic phenotypes exhibited are related to definite microbial profiles in recipients, highlighting the predominant role of Akkermansia genus in the protection from obesity-associated NAFLD development. Conclusions: The results provide scientific support for the prebiotic capacity of quercetin and the transfer of established metabolic profiles through gut microbiota transplantation as a protective strategy against the development of obesity-related NAFLDSIM.V.G.M. and S.S.C. share senior authorship. D.P. and E.N. made equal contribution to the study. D.P., E.N., S.M.F., M.V.G.M., and S.S.C. performed most of the experiments. J.L.O. and F.J. performed statistical analysis. R.J. and J.G.G. assisted for in vivo models. S.S.C. designed the experiments and supervised the study. All the authors wrote the manuscript. The authors thank Drs. Gérard and Rabot, from MICALIS Institute (INRA), for providing germ-free mice. This work was supported by grants from Ministerio de Economía y Competitividad and Fondo Europeo de Desarrollo Regional (FEDER) (BFU2013-48141-R, BFU2017-87960-R), Junta de Castilla y León (LE135U13, GRS 1428/A/16), Junta de Castilla y León and FEDER, (LE063U16), and IIS Hospital La Fe (2017_0092_PP). D.P. was supported by a fellowship from Junta de Castilla y León co-financed by the European Social Fund. E.N. was supported by Fundación de Investigación Sanitaria of León. M.V.G.M. was supported by CIBERehd contracts. CIBERehd is funded by the Instituto de Salud Carlos III, Spai

Protective effect of quercetin treatment on HFD-induced intestinal dysbiosis and barrier dysfunction in an in vivo model of non-alcoholic fatty liver disease

1 p.Gut microbiota is involved in obesity, metabolic syndrome and nonalcoholic fatty liver disease (NAFLD). Quercetin may have the ability to modulate the intestinal microbiota composition, suggesting therapeutic potential in NAFLD. The present study aims to investigate the beneficial effect of quercetin treatment on dysbiosis, intestinal barrier dysfunction and gut-liver axis alteration in high-fat diet (HFD)-fed mice. Resumen de un trabajo resultado del proyecto de investigación financiado por la Consejería de Educación de la Junta de Castilla y León (referencia LE063U16)S