scholarly journals Partially hydrolysed guar gum ameliorates murine intestinal inflammation in association with modulating luminal microbiota and SCFA

2016 ◽  
Vol 116 (7) ◽  
pp. 1199-1205 ◽  
Author(s):  
Tomohisa Takagi ◽  
Yuji Naito ◽  
Yasuki Higashimura ◽  
Chihiro Ushiroda ◽  
Katsura Mizushima ◽  
...  
Keyword(s):  
Real Time ◽  
Intestinal Microbiota ◽  
Real Time Pcr ◽  
Colonic Mucosa ◽  
Guar Gum ◽  
Intestinal Inflammation ◽  
Neutrophil Infiltration ◽  
Water Soluble ◽  
Mucosal Inflammation ◽  
Trinitrobenzene Sulfonic Acid

AbstractPartially hydrolysed guar gum (PHGG), a water-soluble dietary fibre produced by the controlled partial enzymatic hydrolysis of guar gum beans, has various physiological roles. This study aimed to elucidate the beneficial effects of PHGG on colonic mucosal damage in a murine 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis model. Acute colitis was induced in male C57BL/6 mice with TNBS after 2 weeks of pre-feeding with PHGG (5 %). The colonic mucosal inflammation was evaluated using macroscopic damage scores, and neutrophil infiltration was assessed by measuring tissue-associated myeloperoxidase (MPO) activity in the colonic mucosa. TNF-αexpression in the colonic mucosa was measured by ELISA and real-time PCR. Moreover, the intestinal microbiota and production of SCFA were assessed by real-time PCR and HPLC, respectively. Colonic damage due to TNBS administration was significantly ameliorated by PHGG treatment. Furthermore, PHGG significantly inhibited increases in MPO activity and TNF-αprotein and mRNA expression in the colonic mucosa in TNBS-induced colitis. On analysis of intestinal microbiota, we found that the concentration of theClostridium coccoidesgroup (Clostridiumcluster XIVa), theClostridium leptumsubgroup (Clostridiumcluster IV) and theBacteroides fragilisgroup had significantly increased in PHGG-fed mice. On analysis of SCFA, we found that the caecal content of acetic acid, propionic acid and butyric acid had significantly increased in PHGG-fed mice. Together, these results suggest that chronic ingestion of PHGG prevents the development of TNBS-induced colitis in mice by modulating the intestinal microbiota and SCFA, which may be significant in the development of therapeutics for inflammatory bowel disease.

Identification of adeno-associated viral vectors suitable for intestinal gene delivery and modulation of experimental colitis

2012 ◽  
Vol 302 (3) ◽  
pp. G296-G308 ◽  
Author(s):  
Steven Polyak ◽  
Annette Mach ◽  
Stacy Porvasnik ◽  
Lisa Dixon ◽  
Thomas Conlon ◽  
...  
Keyword(s):  
Gene Expression ◽  
Small Intestine ◽  
Gene Transfer ◽  
Real Time ◽  
Real Time Pcr ◽  
Transgene Expression ◽  
Intestinal Inflammation ◽  
Experimental Colitis ◽  
Mucosal Inflammation

Effective gene transfer with sustained gene expression is an important adjunct to the study of intestinal inflammation and future therapy in inflammatory bowel disease. Recombinant adeno-associated virus (AAV) vectors are ideal for gene transfer and long-term transgene expression. The purpose of our study was to identify optimal AAV pseudotypes for transduction of the epithelium in the small intestine and colon, which could be used for studies in experimental colitis. The tropism and transduction efficiencies of AAV pseudotypes 1–10 were examined in murine small intestine and colon 8 wk after administration by real-time PCR and immunohistochemistry. The clinical and histopathological effects of IL-10-mediated intestinal transduction delivered by AAVrh10 were examined in the murine IL-10−/− enterocolitis model. Serum IL-10 levels and IL-10 expression were followed by ELISA and real-time PCR, respectively. AAV pseudotypes 4, 7, 8, 9, and 10 demonstrated optimal intestinal transduction. Transgene expression was sustained 8 wk after administration and was frequently observed in enteroendocrine cells. Long-term IL-10 gene expression and serum IL-10 levels were observed following AAV transduction in an IL-10−/− model of enterocolitis. Animals treated with AAVrh10-IL-10 had lower disease activity index scores, higher colon weight-to-length ratios, and lower microscopic inflammation scores. This study identifies novel AAV pseudotypes with small intestine and colon tropism and sustained transgene expression capable of modulating mucosal inflammation in a murine model of enterocolitis.

scholarly journals Neutrophil Extracellular Traps Impair Intestinal Barrier Function during Experimental Colitis

2020 ◽  
Author(s):  
Elliot Yi-Hsin Lin ◽  
Hsuan-Ju Lai ◽  
Yuan-Kai Cheng ◽  
Kai-Quan Leong ◽  
Li-Chieh Cheng ◽  
...  
Keyword(s):  
Barrier Function ◽  
Intestinal Inflammation ◽  
Intestinal Barrier ◽  
Experimental Colitis ◽  
Neutrophil Extracellular Trap ◽  
Mucosal Barrier ◽  
Mucosal Inflammation ◽  
Intestinal Barrier Function ◽  
Trinitrobenzene Sulfonic Acid ◽  
Barrier Integrity

Aberrant neutrophil extracellular trap (NET) formation and the loss of barrier integrity in inflamed intestinal tissues have long been associated with inflammatory bowel disease (IBD). However, whether NETs alter intestinal epithelium permeability during colitis remains elusive. Here, we demonstrated that NETs promote the breakdown in intestinal barrier function for the pathogenesis of intestinal inflammation in mouse models of colitis. NETs were abundant in the colon of mice with colitis experimentally induced by dextran sulfate sodium (DSS) or 2,4,6-trinitrobenzene sulfonic acid (TNBS). Analysis of the intestinal barrier integrity revealed that NETs impaired gut permeability, enabling the initiation of luminal bacterial translocation and inflammation. Furthermore, NETs induced the apoptosis of epithelial cells and disrupted the integrity of tight junctions and adherens junctions. Intravenous administration of DNase I, an enzyme that dissolves the web-like DNA filaments of NETs, during colitis restored the mucosal barrier integrity which reduced the dissemination of luminal bacteria, and attenuated intestinal inflammation in both DSS and TNBS models. We conclude that NETs serve a detrimental factor in the gut epithelial barrier function leading to the pathogenesis of mucosal inflammation during acute colitis.

scholarly journals Neutrophil Extracellular Traps Impair Intestinal Barrier Function during Experimental Colitis

Biomedicines ◽  
2020 ◽  
Vol 8 (8) ◽  
pp. 275
Author(s):  
Elliot Yi-Hsin Lin ◽  
Hsuan-Ju Lai ◽  
Yuan-Kai Cheng ◽  
Kai-Quan Leong ◽  
Li-Chieh Cheng ◽  
...  
Keyword(s):  
Barrier Function ◽  
Intestinal Inflammation ◽  
Intestinal Barrier ◽  
Experimental Colitis ◽  
Neutrophil Extracellular Trap ◽  
Mucosal Barrier ◽  
Mucosal Inflammation ◽  
Intestinal Barrier Function ◽  
Trinitrobenzene Sulfonic Acid ◽  
Barrier Integrity

Aberrant neutrophil extracellular trap (NET) formation and the loss of barrier integrity in inflamed intestinal tissues have long been associated with inflammatory bowel disease (IBD). However, whether NETs alter intestinal epithelium permeability during colitis remains elusive. Here, we demonstrated that NETs promote the breakdown in intestinal barrier function for the pathogenesis of intestinal inflammation in mouse models of colitis. NETs were abundant in the colon of mice with colitis experimentally induced by dextran sulfate sodium (DSS) or 2,4,6-trinitrobenzene sulfonic acid (TNBS). Analysis of the intestinal barrier integrity revealed that NETs impaired gut permeability, enabling the initiation of luminal bacterial translocation and inflammation. Furthermore, NETs induced the apoptosis of epithelial cells and disrupted the integrity of tight junctions and adherens junctions. Intravenous administration of DNase I, an enzyme that dissolves the web-like DNA filaments of NETs, during colitis restored the mucosal barrier integrity which reduced the dissemination of luminal bacteria and attenuated intestinal inflammation in both DSS and TNBS models. We conclude that NETs serve a detrimental factor in the gut epithelial barrier function leading to the pathogenesis of mucosal inflammation during acute colitis.

scholarly journals Hepatoprotective Effect of Gum Arabic in Non-Alcoholic Fatty Liver and Potential Modulation of Intestinal Microbiota (FS07-07-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Asmaa Ramadan Taha ◽  
Sahar Alokbi ◽  
Hoda Mabrok
Keyword(s):  
Fatty Liver ◽  
Real Time ◽  
Normal Control ◽  
Intestinal Microbiota ◽  
Real Time Pcr ◽  
Gum Arabic ◽  
Hepatoprotective Effect ◽  
Control Group ◽  
Alcoholic Fatty Liver ◽  
Total Fat

Abstract Objectives To study the hepatoprotective effect of gum Arabic in non-alcoholic fatty liver disease, and to test its ability to reverse the disturbance in intestinal microbiota associated with the disease. Methods Eighteen white male albino rats (150 g-170 g) were randomized into 3 groups (6 rats each). Animals of the normal control group were fed on a balanced diet. Non-alcoholic fatty liver was induced in the second group (HF group) by feeding the animals a high fructose-high fat diet (60% fructose,15% fat). Animals of the third group (GA group) were fed on a high fructose-high fat diet containing 10% gum Arabic. After 5 weeks, the animals were sacrificed, their livers were harvested, and feces samples were collected and stored at −80o C until analysis. Liver total fat was estimated using Folch's extraction procedure. Liver sections were prepared and stained for histological examination. Genomic DNA was extracted from feces samples. Two intestinal phyla (Bacteroidetes and Firmicutes) were quantified by real-time PCR using specific primers. Results Estimation of the liver total fat showed a significant increase in the HF group compared to normal control. The GA group showed a significantly lower level of total fat compared to the HF group, which was comparable to the normal control group (P ≥ 0.05). Histological analysis of liver sections revealed severe fatty degeneration in most of the hepatocytes in the HF group. Hepatocytes of the GA group showed a marked improvement which was not far from the normal control. Real-time PCR showed slightly more copies of Bacteroidetes, and a slightly higher ratio of Bacteroidetes to Firmicutes in the GA group compared to the HF group. However, real-time PCR results were not statistically significant at P ≥ 0.05. Conclusions Gum Arabic is a promising functional food with a potential hepatoprotective effect in non-alcoholic fatty liver disease. Gum Arabic did not have a marked modulating effect on intestinal microbiota and more studies are needed in that area. Funding Sources This study was funded by the National Research Center in Egypt. Supporting Tables, Images and/or Graphs

scholarly journals Use of 16S rRNA Gene-Targeted Group-Specific Primers for Real-Time PCR Analysis of Predominant Bacteria in Mouse Feces

2015 ◽  
Vol 81 (19) ◽  
pp. 6749-6756 ◽  
Author(s):  
Yun-Wen Yang ◽  
Mang-Kun Chen ◽  
Bing-Ya Yang ◽  
Xian-Jie Huang ◽  
Xue-Rui Zhang ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Bacterial Species ◽  
Chronic Inflammatory Bowel Disease ◽  
Trinitrobenzene Sulfonic Acid ◽  
Rrna Gene ◽  
Specific Primers ◽  
Content Type

ABSTRACTMouse models are widely used for studying gastrointestinal (GI) tract-related diseases. It is necessary and important to develop a new set of primers to monitor the mouse gut microbiota. In this study, 16S rRNA gene-targeted group-specific primers forFirmicutes,Actinobacteria,Bacteroidetes,Deferribacteres, “CandidatusSaccharibacteria,”Verrucomicrobia,Tenericutes, andProteobacteriawere designed and validated for quantification of the predominant bacterial species in mouse feces by real-time PCR. After confirmation of their accuracy and specificity by high-throughput sequencing technologies, these primers were applied to quantify the changes in the fecal samples from a trinitrobenzene sulfonic acid-induced colitis mouse model. Our results showed that this approach efficiently predicted the occurrence of colitis, such as spontaneous chronic inflammatory bowel disease in transgenic mice. The set of primers developed in this study provides a simple and affordable method to monitor changes in the intestinal microbiota at the phylum level.

scholarly journals Children Living near a Sanitary Landfill Have Increased Breath Methane andMethanobrevibacter smithiiin Their Intestinal Microbiota

Archaea ◽  
2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Humberto Bezerra de Araujo Filho ◽  
Mirian Silva Carmo-Rodrigues ◽  
Carolina Santos Mello ◽  
Lígia Cristina Fonseca Lahoz Melli ◽  
Soraia Tahan ◽  
...  
Keyword(s):  
Real Time ◽  
Intestinal Microbiota ◽  
Real Time Pcr ◽  
Sanitary Landfill ◽  
Socioeconomic Level ◽  
Breath Methane ◽  
Level Group

This study evaluated the breath CH4excretion and concentration ofM. smithiiin intestinal microbiota of schoolchildren from 2 slums. One hundred and eleven children from a slum near a sanitary landfill, 35 children of a slum located away from the sanitary landfill, and 32 children from a high socioeconomic level school were included in the study. Real-time PCR was performed to quantify theM. smithii nifHgene and it was present in the microbiota of all the participating children, with higherP<0.05concentrations in those who lived in the slum near the landfill (3.16×107 CFU/g of feces), comparing with the children from the slum away from the landfill (2.05×106 CFU/g of feces) and those from the high socioeconomic level group (3.93×105 CFU/g of feces). The prevalence of children who present breath methane was 53% in the slum near the landfill, 31% in the slum further away from the landfill and, 22% in the high socioeconomic level group. To live near a landfill is associated with higher concentrations ofM. smithiiin intestinal microbiota, comparing with those who live away from the landfill, regardless of their socioeconomics conditions.

Quantifying protozoal duodenal outflow in steers by real-time PCR

2005 ◽  
Vol 2005 ◽  
pp. 70-70
Author(s):  
D.R. Yáñez Ruiz ◽  
N.D. Scollan ◽  
R.J. Merry ◽  
C.J. Newbold
Keyword(s):  
Amino Acids ◽  
Real Time ◽  
Real Time Pcr ◽  
Molecular Techniques ◽  
Water Soluble ◽  
Soluble Carbohydrates ◽  
Microbial Protein ◽  
Specific Primers ◽  
Water Soluble Carbohydrates ◽  
Routine Procedures

Microbial protein contributes about two-thirds of the amino acids absorbed by ruminants. Information on the proportion of bacterial and protozoal N passing to the duodenum would enhance our understanding of the effect of diet not only on microbial protein synthesis but also on the contribution of bacteria and protozoa to duodenal flow of others metabolites of interest. However, differentiation of the duodenal fractions has proven to be difficult because routine procedures cannot separate microbial protein at the duodenum into bacteria and protozoa (Punia et al., 1992). Molecular techniques that use group-specific rRNA-targeted probes may help overcome these problems. The objective of this experiment was to quantify the flow of protozoal N at the duodenum in steers, fed two silage diets differing in water soluble carbohydrates (WSC) content, by real-time PCR using protozoa specific primers.

scholarly journals Algal Oil Rich in n-3 PUFA Alleviates DSS-Induced Colitis via Regulation of Gut Microbiota and Restoration of Intestinal Barrier

2020 ◽  
Vol 11 ◽  
Author(s):  
Zhenxia Xu ◽  
Hu Tang ◽  
Fenghong Huang ◽  
Zhixian Qiao ◽  
Xu Wang ◽  
...  
Keyword(s):  
Tight Junction ◽  
Intestinal Microbiota ◽  
Intestinal Inflammation ◽  
Intestinal Barrier ◽  
Neutrophil Infiltration ◽  
Tight Junction Proteins ◽  
Mice Model ◽  
Colonic Inflammation ◽  
Algal Oil ◽  
Junction Proteins

Algal oil is rich in docosahexaenoic acid (DHA) and has various health benefits against human metabolic disorders and disease. This study aimed to investigate the effects of DHA algal oil on colonic inflammation and intestinal microbiota in dextran sulfate sodium (DSS)-induced colitis mice model. Male C57BL/6 mice was induced colitis by 2.5% DSS and followed by 2 weeks of treatment with algal oil (250 or 500 mg/kg/day). The colonic inflammation was assessed by colon macroscopic damage scores, and the degree of neutrophil infiltration was evaluated by measuring tissue-associated myeloperoxidase (MPO) activity in colonic mucosa. Tight junction proteins in the colonic tissue were measured by real-time PCR and western blot. Moreover, the intestinal microbiota and shot chain fatty acids (SCFAs) were estimated by bioinformatic analysis and GC, respectively. Colonic damage due to DSS treatment was significantly ameliorated by algal oil supplementation. In addition, algal oil significantly inhibited the increases of malondialdehyde (MDA) content, MPO activity, pro-inflammatory cytokines level and tight junction proteins expression in DSS-treated mice. Furthermore, supplementation of algal oil modulated the intestinal microbiota structure in DSS induced colitis mice by increasing the proportion of the unidentified_S24_7 and decreasing the relative abundance of unidentified_Ruminococcaceae, Clostridium and Roseburia. On the analysis of SCFAs, the caecal content of acetic acid, propionic acid, isobutyric acid, buturic, and the total SCFAs showed a significant increase in algal oil-administered mice. Together, these results suggested that algal oil rich in DHA inhibited the progress of DSS-induced colitis in mice by modulating the intestinal microbiota and metabolites and repairing the intestinal barrier, which may be applied in the development of therapeutics for intestinal inflammation.

Current and future uses of real-time polymerase chain reaction and microarrays in the study of intestinal microbiota, and probiotic use and effectiveness

2007 ◽  
Vol 53 (5) ◽  
pp. 537-550 ◽  
Author(s):  
Christine M. Carey ◽  
Jennifer L. Kirk ◽  
Shivani Ojha ◽  
Magdalena Kostrzynska
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Intestinal Microbiota ◽  
Real Time Pcr ◽  
Gastrointestinal Disease ◽  
Health Benefit ◽  
Nutrient Status ◽  
Atopic Disease ◽  
Chain Reaction ◽  
Polymerase Chain

Probiotics are defined as live microorganisms that confer a health benefit to the host when administered in adequate amounts. In addition to human health benefits, probiotics can improve various aspects of growth and performance in livestock and poultry, as well as control undesirable microorganisms in food animals. Studies indicate that probiotics can prevent or treat certain conditions, including atopic disease in infants, food allergy, infection after surgery, acute diarrhea, and symptoms associated with irritable bowel syndrome. Understanding the complete mechanism, effectiveness, and potential use of probiotics is limited by the availability and sensitivity of current methods (i.e., culturing techniques). In recent years, real-time polymerase chain reaction (PCR) and microarrays have become prominent and promising methods to examine quantitative changes of specific members of the microbial community and the influence of probiotics on the structure and function of human and animal intestinal ecosystems. Culture-independent studies have established that only a fraction of organisms present in feces are cultivable, therefore, results obtained by cultivation are limited. Conversely, in-depth knowledge of microbial genomes has enabled real-time PCR and microarrays to be more sensitive and has resulted in precise methods for comprehensive analysis of the complex gut microbiota. Additionally, these technologies can assess the influence of intestinal microorganisms on host metabolism, nutrient status, and disease. This paper reviews method technologies and applications of real-time PCR and microarray assays as they relate to the effect and use of probiotics on the intestinal microbiota and gastrointestinal disease.

Sign in / Sign up

Export Citation Format

Share Document