scholarly journals Gallic acid affects intestinal-epithelial-cell integrity and selected amino-acid uptake in porcine in vitro and ex vivo permeability models

2020 ◽  
pp. 1-9
Author(s):  
Marco Tretola ◽  
Giuseppe Bee ◽  
Paolo Silacci
Keyword(s):  
Amino Acid ◽  
Epithelial Cell ◽  
Protein Expression ◽  
Gallic Acid ◽  
Intestinal Epithelial Cell ◽  
Ex Vivo ◽  
Amino Acid Transporter ◽  
Intestinal Epithelial ◽  
Cell Integrity

Abstract Gallic acid (GA) is widely used as a dietary supplement due to several health-promoting effects, although its effects on intestinal-epithelial-cell integrity and transport remain mostly unknown. The present study aims to clarify the effects of GA on tight junctions and intestinal nutrient uptake through in vitro and ex vivo models. Both intestinal porcine enterocyte cell line-J2 cells and porcine middle-jejunum segments were treated with 5 (T5), 25 (T25) and 50 (T50) µm GA and mounted in Ussing chambers to determine transepithelial resistance (TEER), claudin-1 (CLDN1), occludin (OCLN), zonula occludens-1 (ZO-1) protein (in tissues and cells) and mRNA (in cells) expression. In addition, uptake of l-glutamate (l-Glut), l-arginine (l-Arg), l-lysine (l-Lys) and l-methionine (l-Meth) together with cationic-amino-acid transporter-1 (CAT-1) and excitatory-amino-acid transporter-3 (EAAT3) expression was evaluated. No apoptosis was observed in GA-treated cells, but TEER and CLDN1 protein abundance was lower with T50 compared with untreated cells. l-Arg and l-Lys uptake was greater with T5 than with T25 and T50. Ex vivo, T50 decreased the TEER values and the protein levels of CLDN1, OCLN and ZO-1, whereas T5 and T25 only decreased CLDN1 protein expression compared with untreated tissues. Moreover, T25 increased l-Glut and l-Arg uptake, the latter confirmed by an increased protein expression of CAT-1. GA influences intestinal uptake of the tested cationic amino acids at low concentrations and decreases the intestinal-cell barrier function at high concentrations. Similarities were observed between in vitro and ex vivo, but different treatment times and structures must be considered.

PSIII-21 Efficacy of deoxynivalenol detoxification by sodium metabisulfite (SMBS) containing nanofibrous mats using an in vitro intestinal epithelial cell IPEC-J2

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 341-342
Author(s):  
Changning Yu ◽  
Quintin Litke ◽  
Peng Lu ◽  
Shangxi Liu ◽  
Joshua Gong ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Intestinal Epithelial Cell ◽  
Gastric Fluid ◽  
Simulated Gastric Fluid ◽  
Sodium Metabisulfite ◽  
Intestinal Epithelial ◽  
Cell Integrity ◽  
Eudragit L100 ◽  
Vitro Analysis

Abstract Deoxynivalenol (DON) occurs in many commonly used cereal grains. Pigs feed with DON concentrations as low as 0.6–2.0 mg/kg can result in reduced feed intake and growth rate, damage to intestinal epithelial cells, and increase susceptibility to enteric pathogens. Sodium metabisulfite (SMBS) can efficiently detoxify DON by converting it 8-DONS or 10-DONS in vitro. However, if SMBS is added directly to the feed, SMBS rapidly degrades under acidic aqueous conditions (e.g. pig stomach) and little SMBS is delivered to the intestinal absorption site where it can effectively detoxify DON. Thus, the objective of this study was to encapsulate SMBS into Eudragit L100-55 nanofibrous mats to deliver intact SMBS to the small intestine and evaluate the efficacy of DON detoxification in the simulated intestine fluid (SIF) using an in vitro intestinal epithelial cell (IPEC-J2) model. Nanofibrous mats were produced by coaxial electrospinning, with peak loading capacity and loading efficiency of SMBS reaching 32.00% and 80.01%. DON-induced cytotoxicity was not observed during in vitro analysis consisting of incubation of DON in the presence of SMBS-containing nanofibers (0.5% w/w) in simulated gastric fluid (SGF) for 2 h followed by incubation in a mixture of SGF and SIF (1:1) for 20 min. Meanwhile, compared to the DON treatment, incubation of DON in the presence of SMBS-containing nanofiber (0.5% w/w) in SGF for 2 h and SIF for 20 min decreased the gene expression of inflammatory cytokines in the IPEC-J2 cells and maintained the cell integrity. To conclude, SMBS released from Eudragit L100-55 nanofibrous mats in the SIF effectively decreased the adverse effects induced by DON in the IPEC-J2 cells. Nanofibrous mats can release a large amount of SMBS in a short time in SIF to achieve the effect of detoxifying DON.

scholarly journals Opinion: Are Organoids the End of Model Evolution for Studying Host Intestinal Epithelium/Microbe Interactions?

2019 ◽  
Vol 7 (10) ◽  
pp. 406 ◽  
Author(s):  
Michelle M. George ◽  
Mushfiqur Rahman ◽  
Jessica Connors ◽  
Andrew W. Stadnyk
Keyword(s):  
Epithelial Cell ◽  
Cell Biology ◽  
Intestinal Epithelial Cell ◽  
Spatial Organization ◽  
Ex Vivo ◽  
Model Systems ◽  
Apical Surface ◽  
Intestinal Epithelial ◽  
Intestinal Organoids

In the pursuit to understand intestinal epithelial cell biology in health and disease, researchers have established various model systems, from whole animals (typically rodents) with experimentally induced disease to transformed human carcinomas. The obvious limitation to the ex vivo or in vitro cell systems was enriching, maintaining, and expanding differentiated intestinal epithelial cell types. The popular concession was human and rodent transformed cells of mainly undifferentiated cells, with a few select lines differentiating along the path to becoming goblet cells. Paneth cells, in particular, remained unculturable. The breakthrough came in the last decade with the report of conditions to grow mouse intestinal organoids. Organoids are 3-dimensional ex vivo “mini-organs” of the organ from which the stem cells were derived. Intestinal organoids contain fully differentiated epithelial cells in the same spatial organization as in the native epithelium. The cells are suitably polarized and produce and secrete mucus onto the apical surface. This review introduces intestinal organoids and provide some thoughts on strengths and weaknesses in the application of organoids to further advance our understanding of the intestinal epithelial–microbe relationship.

scholarly journals Effects of arginine on intestinal epithelial cell integrity and nutrient uptake

2016 ◽  
Vol 116 (10) ◽  
pp. 1675-1681 ◽  
Author(s):  
Mi Xia ◽  
Lulu Ye ◽  
Qihang Hou ◽  
Qinghua Yu
Keyword(s):  
Amino Acid ◽  
Epithelial Cell ◽  
Nutrient Uptake ◽  
Intestinal Epithelial Cell ◽  
Amino Acid Transporter ◽  
Acid Uptake ◽  
Intestinal Epithelial ◽  
Physiological Level ◽  
Acid Transporter ◽  
Arginine Concentration

AbstractArginine is a multifaceted amino acid that is critical to the normal physiology of the gastrointestinal tract. Oral arginine administration has been shown to improve mucosal recovery following intestinal injury. The present study investigated the influence of extracellular arginine concentrations on epithelial cell barrier regulation and nutrition uptake by porcine small intestinal epithelial cell line (IPEC-J2). The results show that reducing arginine concentration from 0·7 to 0·2 mm did not affect the transepithelial electrical resistance value, tight-junction proteins (claudin-1, occludin, E-cadherin), phosphorylated extracellular signal-regulated protein kinases (p-ERK) and mucin-1 expression. Furthermore, reducing arginine concentration stimulated greater expression of cationic amino acid transporter (CAT1), excitatory amino acid transporter (EAAT3) and alanine/serine/cysteine transporter (ASCT1) mRNA by IPEC-J2 cells, which was verified by elevated efficiency of amino acid uptake. Glucose consumption by IPEC-J2 cells treated with 0·2 mm-arginine remained at the same physiological level to guarantee energy supply and to maintain the cell barrier. This experiment implied that reducing arginine concentration is feasible in IPEC-J2 cells guaranteed by nutrient uptake and cell barrier function.

scholarly journals Isoflavones inhibit intestinal epithelial cell proliferation and induce apoptosis in vitro

1999 ◽  
Vol 80 (10) ◽  
pp. 1550-1557 ◽  
Author(s):  
C Booth ◽  
D F Hargreaves ◽  
J A Hadfield ◽  
A T McGown ◽  
C S Potten
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cell ◽  
Intestinal Epithelial Cell ◽  
Intestinal Epithelial ◽  
Epithelial Cell Proliferation

Effects of lactoferrin on intestinal epithelial cell growth and differentiation: an in vivo and in vitro study

BioMetals ◽  
2014 ◽  
Vol 27 (5) ◽  
pp. 857-874 ◽  
Author(s):  
Anne Blais ◽  
Cuibai Fan ◽  
Thierry Voisin ◽  
Najat Aattouri ◽  
Michel Dubarry ◽  
...  
Keyword(s):  
Cell Growth ◽  
Epithelial Cell ◽  
Intestinal Epithelial Cell ◽  
In Vitro Study ◽  
Vitro Study ◽  
Intestinal Epithelial ◽  
Cell Growth And Differentiation

scholarly journals Effects of Ethanol and Acetaldehyde on Tight Junction Integrity: In Vitro Study in a Three Dimensional Intestinal Epithelial Cell Culture Model

PLoS ONE ◽  
2012 ◽  
Vol 7 (4) ◽  
pp. e35008 ◽  
Author(s):  
Elhaseen Elamin ◽  
Daisy Jonkers ◽  
Kati Juuti-Uusitalo ◽  
Sven van IJzendoorn ◽  
Freddy Troost ◽  
...  
Keyword(s):  
Cell Culture ◽  
Epithelial Cell ◽  
Intestinal Epithelial Cell ◽  
Three Dimensional ◽  
In Vitro Study ◽  
Intestinal Epithelial ◽  
Cell Culture Model ◽  
Epithelial Cell Culture ◽  
Culture Model

Heat- and Formalin-Inactivated Probiotic Bacteria Induce Comparable Cytokine Patterns in Intestinal Epithelial Cell–Leucocyte Cocultures

2007 ◽  
Vol 70 (10) ◽  
pp. 2417-2421 ◽  
Author(s):  
ELEONORA DEHLINK ◽  
KONRAD J. DOMIG ◽  
CHRISTINE LOIBICHLER ◽  
ELKE KAMPL ◽  
THOMAS EIWEGGER ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Intestinal Epithelial Cell ◽  
Cytokine Production ◽  
Probiotic Bacteria ◽  
Intestinal Epithelial ◽  
Specific Production ◽  
Peripheral Blood Mononuclear ◽  
Probiotic Strains ◽  
Cytokine Synthesis

The mode of inactivation of probiotic bacteria may profoundly affect their immune-modulatory properties to the point of reversal of effects in in vitro human intestinal epithelial-like cell cultures (Caco-2). To further investigate the influence of inactivation treatment on cytokine production, three probiotic strains were evaluated—live, heat-inactivated, and formalininactivated strains—for their impact on interleukin (IL) 6, IL-8, and IL-10 production in Caco-2–leucocyte cocultures. The tested bacteria induced strain-specific production of IL-6, IL-8, and IL-10. No suppressive effects on cytokine synthesis were observed. Live microorganisms seemed to be slightly more potent inducers of cytokine production than nonviable strains, but differences to inactivated bacteria were not statistically significant. Our results indicate that heat and formalin treatments of probiotic microorganisms are equivalent inactivation methods in terms of induction of IL-6, IL-8, and IL-10 production in Caco-2–peripheral blood mononuclear cell cocultures and do not invert immune-modulatory effects.

scholarly journals Deficiency in the anti-apoptotic protein DJ-1 promotes intestinal epithelial cell apoptosis and aggravates inflammatory bowel disease via p53

2020 ◽  
Vol 295 (13) ◽  
pp. 4237-4251 ◽  
Author(s):  
Jie Zhang ◽  
Min Xu ◽  
Weihua Zhou ◽  
Dejian Li ◽  
Hong Zhang ◽  
...  
Keyword(s):  
Inflammatory Bowel Disease ◽  
Epithelial Cell ◽  
Bowel Disease ◽  
Intestinal Epithelial Cell ◽  
Intestinal Inflammation ◽  
Intestinal Epithelial ◽  
P53 Expression ◽  
Inflammatory Bowel

Parkinson disease autosomal recessive, early onset 7 (PARK7 or DJ-1) is involved in multiple physiological processes and exerts anti-apoptotic effects on multiple cell types. Increased intestinal epithelial cell (IEC) apoptosis and excessive activation of the p53 signaling pathway is a hallmark of inflammatory bowel disease (IBD), which includes ulcerative colitis (UC) and Crohn's disease (CD). However, whether DJ-1 plays a role in colitis is unclear. To determine whether DJ-1 deficiency is involved in the p53 activation that results in IEC apoptosis in colitis, here we performed immunostaining, real-time PCR, and immunoblotting analyses to assess DJ-1 expression in human UC and CD samples. In the inflamed intestines of individuals with IBD, DJ-1 expression was decreased and negatively correlated with p53 expression. DJ-1 deficiency significantly aggravated colitis, evidenced by increased intestinal inflammation and exacerbated IEC apoptosis. Moreover, DJ-1 directly interacted with p53, and reduced DJ-1 levels increased p53 levels both in vivo and in vitro and were associated with decreased p53 degradation via the lysosomal pathway. We also induced experimental colitis with dextran sulfate sodium in mice and found that compared with DJ-1−/− mice, DJ-1−/−p53−/− mice have reduced apoptosis and inflammation and increased epithelial barrier integrity. Furthermore, pharmacological inhibition of p53 relieved inflammation in the DJ-1−/− mice. In conclusion, reduced DJ-1 expression promotes inflammation and IEC apoptosis via p53 in colitis, suggesting that the modulation of DJ-1 expression may be a potential therapeutic strategy for managing colitis.

scholarly journals Absorption of Hemoglobin Iron: The Role of Xanthine Oxidase in the Intestinal Heme-Splitting Reaction

Blood ◽  
1970 ◽  
Vol 35 (1) ◽  
pp. 94-103 ◽  
Author(s):  
R. BEN DAWSON ◽  
SHEILA RAFAL ◽  
LEWIS R. WEINTRAUB
Keyword(s):  
Epithelial Cell ◽  
Xanthine Oxidase ◽  
Intestinal Epithelial Cell ◽  
Oxidase Inhibitor ◽  
Small Intestinal Mucosa ◽  
Intestinal Epithelial ◽  
Sephadex Column ◽  
Xanthine Oxidase Inhibitor

Abstract Heme from ingested hemoglobin—59Fe is taken into the epithelial cell of the small intestinal mucosa of the dog and the 59Fe subsequently appears in the plasma bound to transferrin. A substance was demonstrated in homogenates of the mucosa which releases iron from a hemoglobin substrate in vitro. Thus: (1) The addition of catalase to the mucosal homogenate reduces the "heme-splitting" reaction. In contrast, sodium azide, a catalase inhibitor, potentiates the reaction. This suggests that a peroxide generating system participates in the "heme-splitting" reaction. (2) Xanthine oxidase, an enzyme present in the intestinal epithelial cell, produces H2O2 by oxidation of its substrate. The addition of allopurinol, a xanthine oxidase inhibitor, to the intestinal mucosal homogenate diminishes the "heme-splitting" reaction. (3) Fractionation of the 50,000 Gm. supernatant of the mucosal homogenate on a G-200 Sephadex column shows the "heme-splitting" activity to have the same elution volume as xanthine oxidase, indicating a similar molecular weight. (4) The addition of a mucosal homogenate to a xanthine substrate results in the production of uric acid. These data suggest that xanthine oxidase in the intestinal epithelial cell is important in the release of iron from absorbed heme. The enzyme mediates the "heme-splitting" reaction by the generation of peroxides which, in turn, oxidize the alpha-methene bridge of the heme ring releasing iron and forming biliverdin.

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