Exposure to Hog Dust Extract Decreases Barrier Integrity in Caco‐2 Cells, Induces Airway Inflammation and Increases Intestinal Permeability in Mice

Abstract Objectives 2’-fucosyllactose (2’-FL), the most predominant oligosaccharide found in human milk, acts as a prebiotic with beneficial effects on the host. The aim of this study was to determine the beneficial effect of 2’-FL on intestinal barrier integrity and metabolic functions in low-fat (LF)- and high-fat (HF)-fed mice. Methods Male C57/BL6 mice (n = 32, 8/group; 6 weeks old, JAX, CA) were counter-balanced into four weight-matched groups and fed either a low-fat (LF; 10% kcal fat with 7% kcal sucrose) or HF (45% kcal fat with 17% kcal sucrose) with or without supplementation of 2’-FL in the diet [10% (w/w), 8 weeks; LF/2’-FL or HF/2’-FL; BASF, Germany]. General phenotypes (body weight, energy intake, fat and lean mass), intestinal permeability (ex vivo in Ussing chambers), lipid profiles, and microbial metabolites were assessed. Results 2’-FL significantly attenuated the HF-induced increase in body fat mass with a trend to decrease body weight gain. 2’-FL significantly decreased intestinal permeability in LF-fed mice with a trend for a decrease in HF-fed mice. This was associated with a significant increase in interleukin-22, a cytokine known to have a protective role in intestinal barrier function. Visceral adipocyte size was significantly decreased by 2’-FL in both LF- and HF-fed mice. 2’-FL suppressed HF-induced upregulation of adipogenic transcription factors peroxisome proliferator-activated receptor gamma and sterol regulatory element binding protein-1c in the liver. Lastly, 2’-FL supplementation led to a significant elevation of lactic acid concentration in the cecum of HF-fed mice, which is known to be a product from beneficial microbes. Conclusions 2’-FL supplementation improved gut barrier integrity and lipid metabolism in mice with and without the metabolic challenge of HF feeding. These findings support the use of 2’-FL in the control of gut barrier function and metabolic homeostasis under normal and abnormal physiological conditions. Funding Sources BASF (Germany).

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Functional and structural alterations of epithelial barrier properties of rat ileum following X-irradiation

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y03-129 ◽

2004 ◽

Vol 82 (2) ◽

pp. 84-93 ◽

Cited By ~ 17

Author(s):

I Dublineau ◽

F Lebrun ◽

S Grison ◽

N M Griffiths

Keyword(s):

Intestinal Permeability ◽

Intestinal Barrier ◽

Barrier Properties ◽

Epithelial Tissue ◽

Barrier Integrity ◽

Rat Ileum ◽

Ussing Chambers ◽

Junctional Proteins ◽

X Irradiation

Irradiation of the digestive system leads to alterations of the small intestine. We have characterized the disruption of the barrier integrity in rat ileum from 1 to 14 days following irradiation ranging from 6 to 12 Gy. The intestinal permeability to 14C-mannitol and 3H-dextran 70 000 was measured in vitro in Ussing chambers. In parallel to these functional studies, immunohistochemical analyses of junctional proteins (ZO-1 and β-catenin) of ileal epithelium were performed by confocal microscopy. Irradiation with 10 Gy induced a marked decrease in epithelial tissue resistance at three days and a fivefold increase in mannitol permeability, without modifications of dextran permeability. A disorganization of the localization for ZO-1 and β-catenin was also observed. At 7 days after irradiation, we observed a recovery of the organization of junctional proteins in parallel to a return of intestinal permeability to control value. In addition to these time-dependent effects, a gradual effect on epithelial integrity of the radiation doses was observed 3 days after irradiation. This study shows a disruption of the integrity of the intestinal barrier in rat ileum following abdominal X-irradiation, depending on the time postirradiation and on the delivered dose. The loss of barrier integrity was characterized by a disorganization of proteins of tight and adherent junctions, leading to increased intestinal permeability to mannitol.Key words: intestinal permeability, ZO-1, β-catenin, tight and adherent junctions.

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Palmitic Acid Affects Intestinal Epithelial Barrier Integrity and Permeability In Vitro

Antioxidants ◽

10.3390/antiox9050417 ◽

2020 ◽

Vol 9 (5) ◽

pp. 417

Author(s):

Manuele Gori ◽

Annamaria Altomare ◽

Silvia Cocca ◽

Eleonora Solida ◽

Mentore Ribolsi ◽

...

Keyword(s):

Oxidative Stress ◽

Palmitic Acid ◽

Intestinal Permeability ◽

Adherens Junction ◽

Epithelial Barrier ◽

Epithelial Cell Line ◽

Intestinal Epithelial ◽

Barrier Integrity ◽

And Oxidative Stress

Palmitic acid (PA), a long-chain saturated fatty acid, might activate innate immune cells. PA plays a role in chronic liver disease, diabetes and Crohn’s disease, all of which are associated with impaired intestinal permeability. We investigated the effect of PA, at physiological postprandial intestinal concentrations, on gut epithelium as compared to lipopolysaccharide (LPS) and ethanol, using an in vitro gut model, the human intestinal epithelial cell line Caco-2 grown on transwell inserts. Cytotoxicity and oxidative stress were evaluated; epithelial barrier integrity was investigated by measuring the paracellular flux of fluorescein, and through RT-qPCR and immunofluorescence of tight junction (TJ) and adherens junction (AJ) mRNAs and proteins, respectively. In PA-exposed Caco-2 monolayers, cytotoxicity and oxidative stress were not detected. A significant increase in fluorescein flux was observed in PA-treated monolayers, after 90 min and up to 360 min, whereas with LPS and ethanol, this was only observed at later time-points. Gene expression and immunofluorescence analysis showed TJ and AJ alterations only in PA-exposed monolayers. In conclusion, PA affected intestinal permeability without inducing cytotoxicity or oxidative stress. This effect seemed to be faster and stronger than those with LPS and ethanol. Thus, we hypothesized that PA, besides having an immunomodulatory effect, might play a role in inflammatory and functional intestinal disorders in which the intestinal permeability is altered.

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Inhalation of stable dust extract prevents allergen induced airway inflammation and hyperresponsiveness

Thorax ◽

10.1136/thx.2005.049403 ◽

2006 ◽

Vol 61 (2) ◽

pp. 134-139 ◽

Cited By ~ 51

Author(s):

M Peters

Keyword(s):

Airway Inflammation ◽

Dust Extract

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GPR120 prevents colorectal adenocarcinoma progression by sustaining the mucosal barrier integrity

Scientific Reports ◽

10.1038/s41598-021-03787-7 ◽

2022 ◽

Vol 12 (1) ◽

Author(s):

Federica Rubbino ◽

Valentina Garlatti ◽

Valeria Garzarelli ◽

Luca Massimino ◽

Salvatore Spanò ◽

...

Keyword(s):

Colorectal Cancer ◽

Fatty Acids ◽

Intestinal Permeability ◽

Mucosal Barrier ◽

Phenotypic Characterization ◽

Early Event ◽

Blue Dye ◽

Barrier Integrity ◽

Chain Fatty Acids

AbstractGPR120 (encoded by FFAR4 gene) is a receptor for long chain fatty acids, activated by ω-3 Polyunsaturated Fatty Acids (PUFAs), and expressed in many cell types. Its role in the context of colorectal cancer (CRC) is still puzzling with many controversial evidences. Here, we explored the involvement of epithelial GPR120 in the CRC development. Both in vitro and in vivo experiments were conducted to mimic the conditional deletion of the receptor from gut epithelium. Intestinal permeability and integrity of mucus layer were assessed by using Evans blue dye and immunofluorescence for MUC-2 protein, respectively. Microbiota composition, presence of lipid mediators and short chain fatty acids were analyzed in the stools of conditional GPR120 and wild type (WT) mice. Incidence and grade of tumors were evaluated in all groups of mice before and after colitis-associated cancer. Finally, GPR120 expression was analyzed in 9 human normal tissues, 9 adenomas, and 17 primary adenocarcinomas. Our work for the first time highlights the role of the receptor in the progression of colorectal cancer. We observed that the loss of epithelial GPR120 in the gut results into increased intestinal permeability, microbiota translocation and dysbiosis, which turns into hyperproliferation of epithelial cells, likely through the activation of β -catenin signaling. Therefore, the loss of GPR120 represents an early event of CRC, but avoid its progression as invasive cancer. these results demonstrate that the epithelial GPR120 receptor is essential to maintain the mucosal barrier integrity and to prevent CRC developing. Therefore, our data pave the way to GPR120 as an useful marker for the phenotypic characterization of CRC lesions and as new potential target for CRC prevention.

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Lipid rafts are disrupted in mildly inflamed intestinal microenvironments without overt disruption of the epithelial barrier

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00002.2011 ◽

2012 ◽

Vol 302 (8) ◽

pp. G781-G793 ◽

Cited By ~ 18

Author(s):

Rachel V. Bowie ◽

Simona Donatello ◽

Clíona Lyes ◽

Mark B. Owens ◽

Irina S. Babina ◽

...

Keyword(s):

Lipid Rafts ◽

Intestinal Permeability ◽

Barrier Function ◽

Epithelial Barrier ◽

Intestinal Epithelial ◽

Intestinal Epithelial Barrier ◽

Epithelial Barrier Function ◽

Barrier Integrity ◽

Ifn Γ

Intestinal epithelial barrier disruption is a feature of inflammatory bowel disease (IBD), but whether barrier disruption precedes or merely accompanies inflammation remains controversial. Tight junction (TJ) adhesion complexes control epithelial barrier integrity. Since some TJ proteins reside in cholesterol-enriched regions of the cell membrane termed lipid rafts, we sought to elucidate the relationship between rafts and intestinal epithelial barrier function. Lipid rafts were isolated from Caco-2 intestinal epithelial cells primed with the proinflammatory cytokine interferon-γ (IFN-γ) or treated with methyl-β-cyclodextrin as a positive control for raft disruption. Rafts were also isolated from the ilea of mice in which colitis had been induced in conjunction with in vivo intestinal permeability measurements, and lastly from intestinal biopsies of ulcerative colitis (UC) patients with predominantly mild or quiescent disease. Raft distribution was analyzed by measuring activity of the raft-associated enzyme alkaline phosphatase and by performing Western blot analysis for flotillin-1. Epithelial barrier integrity was estimated by measuring transepithelial resistance in cytokine-treated cells or in vivo permeability to fluorescent dextran in colitic mice. Raft and nonraft fractions were analyzed by Western blotting for the TJ proteins occludin and zonula occludens-1 (ZO-1). Our results revealed that lipid rafts were disrupted in IFN-γ-treated cells, in the ilea of mice with subclinical colitis, and in UC patients with quiescent inflammation. This was not associated with a clear pattern of occludin or ZO-1 relocalization from raft to nonraft fractions. Significantly, a time-course study in colitic mice revealed that disruption of lipid rafts preceded the onset of increased intestinal permeability. Our data suggest for the first time that lipid raft disruption occurs early in the inflammatory cascade in murine and human colitis and, we speculate, may contribute to subsequent disruption of epithelial barrier function.

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IL-10 reduces grain dust-induced airway inflammation and airway hyperreactivity

Journal of Applied Physiology ◽

10.1152/jappl.2000.88.1.173 ◽

2000 ◽

Vol 88 (1) ◽

pp. 173-179 ◽

Cited By ~ 13

Author(s):

Timothy J. Quinn ◽

Somer Taylor ◽

Christine L. Wohlford-Lenane ◽

David A. Schwartz

Keyword(s):

Airway Inflammation ◽

Airway Disease ◽

Lavage Fluid ◽

Interleukin 10 ◽

Airway Hyperreactivity ◽

Lower Percentage ◽

Polymorphonuclear Cells ◽

Inhalation Challenge ◽

Grain Dust ◽

Dust Extract

To determine whether interleukin-10 (IL-10) could alter the development of grain dust-induced airway disease, we pretreated mice with either saline or IL-10 intravenously, exposed the mice to an inhalation challenge with corn dust extract (CDE), and measured inflammation and the development of airway hyperreactivity. Pretreatment with IL-10, in comparison to saline, reduced the concentration and percentage of polymorphonuclear cells in the lavage fluid 30 min after the inhalation challenge with CDE ( P < 0.05). In comparison to saline-treated mice, IL-10 did not significantly alter the degree of airway hyperreactivity 30 min after the exposure to CDE. IL-10-treated mice lavaged 18 h after challenge with CDE also exhibited a lower percentage of polymorphonuclear cells in the lavage fluid ( P < 0.05) and had significantly less airway hyperreactivity than did mice pretreated with the saline placebo ( P < 0.05). These findings indicate that exogenous IL-10 is effective in reducing airway inflammation and airway hyperreactivity due to the inhalation of CDE.

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