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Modulation of Intestinal Epithelial Barrier Function by Rhamnogalacturonan

2017· article· en· W4389021946 on OpenAlexafffundabout
Cristiane Hatsuko Baggio, Fernando Lopes, Adamara Machado Nascimento, Thales R. Cipriani, Derek M. McKay, Wallace K. MacNaughton

Bibliographic record

VenueThe FASEB Journal · 2017
Typearticle
Languageen
FieldAgricultural and Biological Sciences
TopicPolysaccharides and Plant Cell Walls
Canadian institutionsUniversity of Calgary
FundersUniversity of Calgary
KeywordsBarrier functionTight junctionCalciumDextranEGTAChemistryCaco-2Ussing chamberCell biologyBiochemistryIn vitroBiology

Abstract

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Background The Inflammatory Bowel Diseases (IBD) are chronic inflammatory disorders of the gastrointestinal tract and are associated with changes in intestinal epithelial barrier function. However, despite several effective therapeutic options, many patients become refractory to the treatment and adverse effects have been reported. For this reason, the development of new drugs targeting the epithelial barrier function is essential. Recently, polysaccharides isolated from plants and fungi have emerged as an important class of natural products with immunomodulatory, anti‐inflammatory and wound healing activity. Previous studies showed that rhamnogalacturonan (RGal), a polysaccharide isolated from Acmella oleracea , protected mice against colitis induced by dextran sulfate sodium (DSS). However, the exact mechanism of action of RGal are not fully clarified. We hypothesized that RGal modulates the intestinal epithelial barrier function and assessed the signaling pathways involved in RGal effect. Methods Human colonic epithelial Caco‐2 cells were seeded on Snapwells, grown for 6 days, and mounted in Ussing chambers to measure the epithelial barrier function [transepithelial electrical resistance (TER) and FITC‐dextran permeability]. Cells were apically treated with RGal (10–1000 μg/ml) after 30 min of FITC‐dextran addition. In a calcium‐switch assay, the epithelial monolayers were temporarily exposed to zero calcium conditions (calcium‐free buffer containing the calcium chelator, EGTA) to disassemble tight junctions. RGal (10–1000 μg/ml) was added during calcium‐free conditions. The FITC‐dextran permeability was also evaluated in Caco‐2 cells stimulated with IFNg plus TNF‐a (25 ng/ml, basolateral) and treated with RGal (1000 μg/ml, apical) every 24 h for 48 h. To evaluate the signaling pathways, the cells were pre‐treated with selective inhibitors of MEK1/2, p38 or PI3K 30 minutes before RGal addition. Results Apical addition of RGal at concentration of 1000 μg/ml significantly increased TER in 185% and caused a 64% reduction in FITC‐dextran flux compared to control group. In addition, RGal also was able to accelerate the tight junction reassembly in 155% during a reversible barrier disruption induced by calcium depletion. When the barrier disruption was caused by cytokines, RGal (1000 μg/ml) significantly reduced the FITC‐dextran flux by 33 and 24% at 30 and 60 min, respectively. However, the signaling pathway inhibitors did not reverse the RGal effect on TER suggesting that MAPkinase and PI3K pathways were not involved. Conclusions These findings demonstrated that RGal increases intestinal epithelial barrier function which appears to be due to effects on tight junction assembly. Further experiments will be carried out to determine the tight junction proteins and signaling pathways are involved in the RGal effect. Elucidation of RGal mechanisms of action could lead to new approaches to the treatment of IBD. Support or Funding Information Dr. Baggio is supported by an Eyes High postdoctoral fellowship from the University of Calgary. Her research project is funded by a grant from Crohn's and Colitis Canada.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.001
Threshold uncertainty score0.002

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0010.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.019
GPT teacher head0.213
Teacher spread0.194 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

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Citations1
Published2017
Admission routes3
Has abstractyes

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