Assessing Mixture Effects of Cereulide and Deoxynivalenol on Intestinal Barrier Integrity and Uptake in Differentiated Human Caco-2 Cells

The human intestine is regularly exposed to ingested food contaminants, such as fungal and bacterial toxins, which have been described to co-occur in a mixed diet. Thus, it is of utmost importance to understand possible interactions between contaminants of different origin. Hence, we investigated the single and combined effects of one of the most abundant mycotoxins, deoxynivalenol (DON; 0.1 to 10 µg/mL), and the bacterial toxin cereulide (CER; 1 to 100 ng/mL) on differentiated human Caco-2 (C2BBe1) cells cultured in a transwell system. We tested the capacity of the two toxins to alter the intestinal integrity and further investigated the uptake of both compounds and the formation of selected DON metabolites. CER alone (10 and 100 ng/mL) and in combination with DON (10 ng/mL CER with 1 µg/mL DON) was found to alter the barrier function by increasing the transepithelial electrical resistance and the expression of the tight junction protein claudin-4. For the first time, DON-3-sulfate was identified as a metabolite of human intestinal cells in vitro. Moreover, co-incubation of CER and DON led to an altered ratio between DON and DON-3-sulfate. Hence, we conclude that co-exposure to CER and DON may alter the intestinal barrier function and biotransformation of intestinal cells.

Download Full-text

Clostridium Butyricum ZJU-F1 Benefits the Intestinal Barrier Function and Immune Response Associated with Its Modulation of Gut Microbiota in Weaned Piglets

Cells ◽

10.3390/cells10030527 ◽

2021 ◽

Vol 10 (3) ◽

pp. 527

Author(s):

Jie Fu ◽

Tenghao Wang ◽

Xiao Xiao ◽

Yuanzhi Cheng ◽

Fengqin Wang ◽

...

Keyword(s):

Immune Response ◽

Bacillus Licheniformis ◽

Barrier Function ◽

Intestinal Barrier ◽

Clostridium Butyricum ◽

Intestinal Barrier Function ◽

Weaned Piglets ◽

Total Acid ◽

Junction Protein ◽

Mucin Expression

This study investigated the effects of dietary C. butyricum ZJU-F1 on the apparent digestibility of nutrients, intestinal barrier function, immune response, and microflora of weaned piglets, with the aim of providing a theoretical basis for the application of Clostridium butyricum as an alternative to antibiotics in weaned piglets. A total of 120 weanling piglets were randomly divided into four treatment groups, in which piglets were fed a basal diet supplemented with antibiotics (CON), Bacillus licheniformis (BL), Clostridium butyricum ZJU-F1 (CB), or Clostridium butyricum and Bacillus licheniformis (CB-BL), respectively. The results showed that CB and CB-BL treatment increased the intestinal digestibility of nutrients, decreased intestinal permeability, and increased intestinal tight junction protein and mucin expression, thus maintaining the integrity of the intestinal epithelial barrier. CB and CB-BL, as exogenous probiotics, were also found to stimulate the immune response of weaned piglets and improve the expression of antimicrobial peptides in the ileum. In addition, dietary CB and CB-BL increased the proportion of Lactobacillus. The levels of butyric acid, propionic acid, acetic acid, and total acid were significantly increased in the ceca of piglets fed CB and CB-BL. Furthermore, we validated the effects of C. butyricum ZJU-F1 on the intestinal barrier function and immune response in vitro and found C. butyricum ZJU-F1 improved intestinal function and enhanced the TLR-2-MyD88-NF-κB signaling.

Download Full-text

The Effects of Sulfated Secondary Bile Acids on Intestinal Barrier Function and Immune Response in an Inflammatory in vitro Human Intestinal Model

SSRN Electronic Journal ◽

10.2139/ssrn.3940829 ◽

2021 ◽

Author(s):

Benthe van der Lugt ◽

Maartje C.P. Vos ◽

Mechteld Grootte Bromhaar ◽

Noortje Ijssennagger ◽

Frank Vrieling ◽

...

Keyword(s):

Immune Response ◽

Bile Acids ◽

Barrier Function ◽

Intestinal Barrier ◽

Intestinal Barrier Function ◽

Secondary Bile Acids

Download Full-text

The mRNA-binding protein IGF2BP1 maintains intestinal barrier function by up-regulating occludin expression

Journal of Biological Chemistry ◽

10.1074/jbc.ac120.013646 ◽

2020 ◽

Vol 295 (25) ◽

pp. 8602-8612

Author(s):

Vikash Singh ◽

Chethana P. Gowda ◽

Vishal Singh ◽

Ashwinkumar S. Ganapathy ◽

Dipti M. Karamchandani ◽

...

Keyword(s):

Intestinal Epithelium ◽

Barrier Function ◽

Binding Protein ◽

Intestinal Barrier ◽

Intestinal Barrier Function ◽

Intestinal Epithelial ◽

Adult Mice ◽

Junction Protein ◽

Mrna Binding Protein ◽

Mrna Binding

Insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) is an mRNA-binding protein that has an oncofetal pattern of expression. It is also expressed in intestinal tissue, suggesting that it has a possible role in intestinal homeostasis. To investigate this possibility, here we generated Villin CreERT2:Igf2bp1flox/flox mice, which enabled induction of an IGF2BP1 knockout specifically in intestinal epithelial cells (IECs) of adult mice. Using gut barrier and epithelial permeability assays and several biochemical approaches, we found that IGF2BP1 ablation in the adult intestinal epithelium causes mild active colitis and mild-to-moderate active enteritis. Moreover, the IGF2BP1 deletion aggravated dextran sodium sulfate–induced colitis. We also found that IGF2BP1 removal compromises barrier function of the intestinal epithelium, resulting from altered protein expression at tight junctions. Mechanistically, IGF2BP1 interacted with the mRNA of the tight-junction protein occludin (Ocln), stabilizing Ocln mRNA and inducing expression of occludin in IECs. Furthermore, ectopic occludin expression in IGF2BP1-knockdown cells restored barrier function. We conclude that IGF2BP1-dependent regulation of occludin expression is an important mechanism in intestinal barrier function maintenance and in the prevention of colitis.

Download Full-text

Lubiprostone Induces Claudin-1 and Protects Intestinal Barrier Function

Pharmacology ◽

10.1159/000503054 ◽

2019 ◽

Vol 105 (1-2) ◽

pp. 102-108 ◽

Cited By ~ 1

Author(s):

Norio Nishii ◽

Tadayuki Oshima ◽

Min Li ◽

Hirotsugu Eda ◽

Kumiko Nakamura ◽

...

Keyword(s):

Barrier Function ◽

Intestinal Barrier ◽

Fluorescein Isothiocyanate ◽

Intestinal Barrier Function ◽

Epithelial Barrier ◽

Dependent Manner ◽

Epithelial Permeability ◽

Epithelial Barrier Function ◽

Dose Dependent

Introduction: Lubiprostone, a chloride channel activator, is said to reduce epithelial permeability. However, whether lubiprostone has a direct effect on the epithelial barrier function and how it modulates the intestinal barrier function remain unknown. Therefore, the effects of lubiprostone on intestinal barrier function were evaluated in vitro. Methods: Caco-2 cells were used to assess the intestinal barrier function. To examine the expression of claudins, immunoblotting was performed with specific antibodies. The effects of lubiprostone on cytokines (IFNγ, IL-6, and IL-1β) and aspirin-induced epithelial barrier disruption were assessed by transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC) labeled-dextran permeability. Results: IFNγ, IL-6, IL-1β, and aspirin significantly decreased TEER and increased epithelial permeability. Lubiprostone significantly improved the IFNγ-induced decrease in TEER in a dose-dependent manner. Lubiprostone significantly reduced the IFNγ-induced increase in FITC labeled-dextran permeability. The changes induced by IL-6, IL-1β, and aspirin were not affected by lubiprostone. The expression of claudin-1, but not claudin-3, claudin-4, occludin, and ZO-1 was significantly increased by lubiprostone. Conclusion: Lubiprostone significantly improved the IFNγ-induced decrease in TEER and increase in FITC labeled-dextran permeability. Lubiprostone increased the expression of claudin-1, and this increase may be related to the effect of lubiprostone on the epithelial barrier function.

Download Full-text

Role for diet in normal gut barrier function: developing guidance within the framework of food-labeling regulations

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00063.2019 ◽

2019 ◽

Vol 317 (1) ◽

pp. G17-G39 ◽

Cited By ~ 10

Author(s):

Michael Camilleri ◽

Barbara J. Lyle ◽

Karen L. Madsen ◽

Justin Sonnenburg ◽

Kristin Verbeke ◽

...

Keyword(s):

Barrier Function ◽

Intestinal Barrier ◽

Normal Function ◽

Model Systems ◽

Intestinal Barrier Function ◽

Dietary Interventions ◽

Labeling Regulations ◽

Gut Barrier Function ◽

And Function

A reduction in intestinal barrier function is currently believed to play an important role in pathogenesis of many diseases, as it facilitates passage of injurious factors such as lipopolysaccharide, peptidoglycan, whole bacteria, and other toxins to traverse the barrier to damage the intestine or enter the portal circulation. Currently available evidence in animal models and in vitro systems has shown that certain dietary interventions can be used to reinforce the intestinal barrier to prevent the development of disease. The relevance of these studies to human health is unknown. Herein, we define the components of the intestinal barrier, review available modalities to assess its structure and function in humans, and review the available evidence in model systems or perturbations in humans that diet can be used to fortify intestinal barrier function. Acknowledging the technical challenges and the present gaps in knowledge, we provide a conceptual framework by which evidence could be developed to support the notion that diet can reinforce human intestinal barrier function to restore normal function and potentially reduce the risk for disease. Such evidence would provide information on the development of healthier diets and serve to provide a framework by which federal agencies such as the US Food and Drug Administration can evaluate evidence linking diet with normal human structure/function claims focused on reducing risk of disease in the general public.

Download Full-text

Betaine attenuates LPS-induced downregulation of Occludin and Claudin-1 and restores intestinal barrier function

10.21203/rs.2.18826/v2 ◽

2020 ◽

Author(s):

Jingtao Wu ◽

Caimei He ◽

Jie Bu ◽

Yue Luo ◽

Shuyuan Yang ◽

...

Keyword(s):

Barrier Function ◽

Intestinal Barrier ◽

Protective Role ◽

Vital Role ◽

Inflammatory Factors ◽

Intestinal Barrier Function ◽

Epithelial Barrier ◽

Epithelial Barriers ◽

Vitro Model

Abstract Background：The intestinal epithelial barrier, which works as the first line of defense between the luminal environment and the host, once destroyed, it will cause serious inflammation or other intestinal diseases. Tight junctions (TJs) play a vital role to maintain the integrity of the epithelial barrier. Lipopolysaccharide (LPS), one of the most important inflammatory factors will downregulate specific TJ proteins including Occludin and Claudin-1 and impair integrity of the epithelial barrier. Betaine has excellent anti-inflammatory activity but whether betaine has any effect on TJ proteins, particularly on LPS-induced dysfunction of epithelial barriers remains unknown. The purpose of this study is to explore the pharmacological effect of betaine on improving intestinal barrier function represented by TJ proteins. Intestinal porcine epithelial cells (IPEC-J2) were used as an in vitro model. Results: The results demonstrated that betaine enhanced the expression of TJ proteins while LPS (1µg/mL) downregulates the expression of these proteins. Furthermore, betaine attenuates LPS-induced decreases of TJ proteins both shown by Western blot (WB) and Reverse transcription- polymerase chain reaction (RT-PCR). The immunofluorescent images consistently revealed that LPS induced the disruption of TJ protein Claudin-1 and reduced its expression while betaine could reverse these alterations. Similar protective role of betaine on intestinal barrier function was observed by transepithelial electrical resistance (TEER) approach. Conclusion: In conclusion, our research demonstrated that betaine attenuated LPS-induced downregulation of Occludin and Claudin-1 and restored the intestinal barrier function.

Download Full-text

Galli gigeriae endothelium corneum: its intestinal barrier protective activity in vitro and chemical composition

Chinese Medicine ◽

10.1186/s13020-021-00432-3 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Shanshan Li ◽

Meng Zheng ◽

Zhentang Zhang ◽

Hengying Peng ◽

Wenling Dai ◽

...

Keyword(s):

Chemical Composition ◽

Mrna Expression ◽

Barrier Function ◽

Intestinal Barrier ◽

Raw264.7 Cells ◽

Intestinal Barrier Function ◽

Dependent Manner ◽

Chemical Profile ◽

Tnf Α ◽

Junction Protein

Abstract Background Galli gigeriae endothelium corneum (GGEC) has been effectively used for centuries for the treatment of functional dyspepsia (FD) in clinical practice in Asian countries. However, its potential mechanism and chemical composition remains undertermined. Methods In this study, the chemical profile of GGEC ethyl acetate extract (EAE) was evaluated by HPLC-Q-TOF–MS/MS. The effects of EAE on intestinal barrier function and inflammation were investigated in IEC-6 cells and RAW264.7 cells. Results The results showed that 33 compounds were tentatively identified, including 12 soy isoflavones, 7 bile acids for the first time in EAE. EAE significantly reinforced intestinal barrier function via increasing the tight junction protein levels of ZO-1 and Occludin, reducing the mRNA expression levels of interleukin (IL)-1β and IL-6 in tumor necrosis factor alpha (TNF-α)-challenged IEC-6 cells. The scratch wound assay showed that EAE accelerated wound healing of IEC-6 cells. EAE evidently reduced the level of NO in a dose-dependent manner with an IC50 value of 18.12 μg/mL, and the mRNA expression of TNF-α, IL-1β, IL-6, iNOS and COX-2 in LPS-treated RAW264.7 cells. Conclusion This study revealed the intestinal barrier protective effects and chemical profile of GGEC, and the results indicated that GGEC strengthened the intestinal barrier by up-regulating protein expression of tight junctions and limiting inflammatory responses.

Download Full-text

Aflatoxin B1 and Aflatoxin M1 Induce Compromised Intestinal Integrity through Clathrin-Mediated Endocytosis

Toxins ◽

10.3390/toxins13030184 ◽

2021 ◽

Vol 13 (3) ◽

pp. 184

Author(s):

Yanan Gao ◽

Xiaoyu Bao ◽

Lu Meng ◽

Huimin Liu ◽

Jiaqi Wang ◽

...

Keyword(s):

Aflatoxin B1 ◽

Intestinal Barrier ◽

Fluorescein Isothiocyanate ◽

Intestinal Barrier Function ◽

Aflatoxin M1 ◽

Dependent Manner ◽

Simultaneous Exposure ◽

Intestinal Integrity

With the growing diversity and complexity of diet, humans are at risk of simultaneous exposure to aflatoxin B1 (AFB1) and aflatoxin M1 (AFM1), which are well-known contaminants in dairy and other agricultural products worldwide. The intestine represents the first barrier against external contaminants; however, evidence about the combined effect of AFB1 and AFM1 on intestinal integrity is lacking. In vivo, the serum biochemical parameters related to intestinal barrier function, ratio of villus height/crypt depth, and distribution pattern of claudin-1 and zonula occluden-1 were significantly affected in mice exposed to 0.3 mg/kg b.w. AFB1 and 3.0 mg/kg b.w. AFM1. In vitro results on differentiated Caco-2 cells showed that individual and combined AFB1 (0.5 and 4 μg/mL) and AFM1 (0.5 and 4 μg/mL) decreased cell viability and trans-epithelial electrical resistance values as well as increased paracellular permeability of fluorescein isothiocyanate-dextran in a dose-dependent manner. Furthermore, AFM1 aggravated AFB1-induced compromised intestinal barrier, as demonstrated by the down-regulation of tight junction proteins and their redistribution, particularly internalization. Adding the inhibitor chlorpromazine illustrated that clathrin-mediated endocytosis partially contributed to the compromised intestinal integrity. Synergistic and additive effects were the predominant interactions, suggesting that these toxins are likely to have negative effects on human health.

Download Full-text

PSX-A-9 Late-Breaking: Effects of farm noise on ileum intestinal barrier function of pullets

Journal of Animal Science ◽

10.1093/jas/skab235.673 ◽

2021 ◽

Vol 99 (Supplement_3) ◽

pp. 367-367

Author(s):

Chun Li ◽

Runxiang Zhang ◽

Hanlin Yu ◽

Yanru Feng ◽

Jianhong Li ◽

...

Keyword(s):

Barrier Function ◽

Intestinal Barrier ◽

Animal Husbandry ◽

Low Noise ◽

Intestinal Barrier Function ◽

Control Group ◽

Tissue Samples ◽

Real Time Quantitative Pcr ◽

Junction Protein ◽

Related Proteins

Abstract Noise is a potential but not negligible environmental factor in animal husbandry. To investigate the effects of farm noise on intestinal barrier function of pullets, 336 Hailanhe pullets aged 1 day were randomly divided into 3 groups: control group (CON), low noise group (LN), high noise group (HN). LN group and HN group were exposed to noise respectively at 65–75 dB and 85–95 dB, the average and the range of the highest loudness of noise in laying hens’ farms for 6h every day (7:00-19:00, hourly intervals for one hour) and lasted 4 weeks. Non additional noise addition in CON group, noise loudness of which was less than 40dB. 6 birds were randomly chosen form each group after every week of noise stimulation for ileum tissue samples. Hematoxylin-eosin stain (HE stain), immunofluorescence, and real-time quantitative PCR (qRT-PCR) were used to determine changes in ileum structure, expression of intestinal barrier related proteins and mRNAs and HSPs. Results shown that 1 week and 2 weeks after noise exposed inflammatory cell infiltration reduced, the expression of intestinal barrier related proteins (Occludin, Mucin2 and ZO-1) and mRNAs (Claudin-1, Claudin-4, E-cadherin, Occludin, Mucin2, ZO-1 and ZO-2) were significantly increased (P < 0.05), the mRNA expression of HSPs decreased (P < 0.05) or have no significate changes (P > 0.05). After 4 weeks of noise treatment, the expression of mRNAs of intestinal tight junction protein and mucin, HSPs were significantly decreased (P < 0.05). There was no difference between the LN and HN groups on those indicators (P > 0.05). The study indicates that noise at 65-75dB and 85-95dB does not cause stress to ileum of pullets while promote the development of intestinal barrier of chicks within 2 weeks maybe by mild stimulation and birds restored to balance due to habitualization after 4 weeks of noise treatment.

Download Full-text