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 62 Evaluating detoxification efficacy of deoxynivalenol by encapsulated sodium metabisulfite (SMBS) using an in vitro intestinal epithelial cell IPEC-J2 model

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 38-39
Author(s):  
Peng Lu ◽  
Changning Yu ◽  
Shangxi Liu ◽  
Joshua Gong ◽  
Song Liu ◽  
...  
Keyword(s):  
Small Intestine ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cell ◽  
Intestinal Epithelial Cells ◽  
In Vitro Release ◽  
Simulated Gastric Fluid ◽  
Sodium Metabisulfite ◽  
Intestinal Epithelial

Abstract Deoxynivalenol (DON) contamination occurs on feed ingredients and causes a reduction in growth performance, damage to the intestinal epithelial cells, and increased susceptibility to enteric pathogen challenge. Sodium metabisulfite (SMBS) has been successfully used to destroy DON in processed grains or feeds. However, SMBS degrades quickly under aqueous acid conditions, such as pig stomachs, and when SMBS is added to diet, little will remain intact in the small intestine where an optimal pH environment exists for detoxification by SMBS. Thus, this study was to encapsulate SMBS into microparticles to deliver intact SMBS to the small intestine and evaluate its efficacy of DON detoxification in the simulated intestine fluid (SIF) using an in vitro intestinal epithelial cell (IPEC-J2) model. The results showed that around 40% of the SMBS loading capacity was achieved in the microparticles. In vitro release studies showed that 1.61% of encapsulated SMBS was released in the simulated gastric fluid (SGF), and the majority of encapsulated SMBS (75.52%) was progressively released in the SIF within 6 h at 37 °C. In vitro cell experiments showed that DON treated with the SIF containing 0.5% SMBS for 2 h completely attenuated the DON-induced cytotoxicity. When DON was treated with the SGF containing 0.5% encapsulated SMBS for 2 h and then the mixture was mixed with the SIF (1:1) and incubated for 2 h, it also completely attenuated the DON-induced cytotoxicity. Moreover, DON treated with the simulated fluid containing 0.5% encapsulated SMBS completely attenuated the gene expression inflammatory cytokines upregulated by DON and restored trans-epithelial electrical resistance (TEER) and tight junction and cytoskeleton. In summary, the encapsulation of SMBS was stable in SGF and allowed a progressive release of SMBS in the SIF. Moreover, the released SMBS in the SIF effectively attenuated the adverse effects induced by DON in the intestinal epithelial cells.

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.

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.

Human intestinal epithelial cell survival: differentiation state-specific control mechanisms

2001 ◽  
Vol 280 (6) ◽  
pp. C1540-C1554 ◽  
Author(s):  
Rémy Gauthier ◽  
Charlène Harnois ◽  
Jean-François Drolet ◽  
John C. Reed ◽  
Anne Vézina ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Survival ◽  
Intestinal Epithelial Cell ◽  
The State ◽  
Control Mechanisms ◽  
Intestinal Epithelial ◽  
Human Intestinal Epithelial Cell ◽  
Expression Levels ◽  
Specific Control

To investigate whether human intestinal epithelial cell survival involves distinct control mechanisms depending on the state of differentiation, we analyzed the in vitro effects of insulin, pharmacological inhibitors of Fak, MEK/Erk, and PI3-K/Akt, and integrin (β1, β4)-blocking antibodies on the survival of the well-established human Caco-2 enterocyte-like and HIEC-6 cryptlike cell models. In addition, relative expression levels of six Bcl-2 homologs (Bcl-2, Bcl-XL, Mcl-1, Bax, Bak, and Bad) and activation levels of Fak, Erk-2, and Akt were analyzed. Herein, we report that 1) the enterocytic differentiation process results in the establishment of distinct profiles of Bcl-2 homolog expression levels, as well as p125Fak, p42Erk-2, and p57Aktactivated levels; 2) the inhibition of Fak, of the MEK/Erk pathway, or of PI3-K, have distinct impacts on enterocytic cell survival in undifferentiated (subconfluent Caco-2, confluent HIEC-6) and differentiated (30 days postconfluent Caco-2) cells; 3) exposure to insulin and the inhibition of Fak, MEK, and PI3-K resulted in differentiation state-distinct modulations in the expression of each Bcl-2 homolog analyzed; and 4) Fak, β1 and β4 integrins, as well as the MEK/Erk and PI3-K/Akt pathways, are distinctively involved in cell survival depending on the state of cell differentiation. Taken together, these data indicate that human intestinal epithelial cell survival is regulated according to differentiation state-specific control mechanisms.

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