RGal Increases Intestinal Epithelial Barrier Function in a PKC‐Dependent Manner and Drives an Inflammatory Gene Expression Profile
Notice bibliographique
Résumé
Background Loss of intestinal epithelial barrier function has been linked to inflammatory bowel disease (IBD). We previously showed that the dietary fibre, rhamnogalacturonan (RGal), reduces inflammation in a murine model of DSS colitis. Additionally, apical treatment of Caco2 human intestinal epithelial cells with RGal increases transepithelial electrical resistance (TER) and accelerates wound‐healing, but the underlying mechanisms remain unknown. Aims We aimed to determine the mechanisms of the RGal mediated increase in epithelial barrier function. (1) Given the ability of PKC to modulate barrier permeability we aimed to determine the role of PKC in the RGal‐mediated increase in epithelial barrier function. (2) We aimed to determine changes in gene expression following RGal treatment. Methods (1) To determine the role of PKC in the RGal‐induced increase in epithelial barrier function, Caco2 monolayers were mounted in Ussing Chambers, pre‐treated with pan protein kinase C (PKC), PKCz‐specific, or classical PKC inhibitors (GFX, PKCz pseudosubstrate or Go6976 respectively) and then treated with RGal. Change in TER and FITC‐dextran flux in response to RGal were assessed. (2) To determine the transcriptionally dependent effects of RGal on epithelial barrier function, Caco2 monolayers were treated apically for 6, 12 and 24h with RGal and RNAseq was performed. Genes with changes in expression more than 3‐fold were entered into Enrichr for pathway analysis. In order to confirm changes in gene expression from RNAseq, a multi‐array immunoassay was performed. Results (1) RGal (1 mg/mL) reduced FITC‐dextran flux by 52.0% (n=5, p<0.05) 30 min post‐treatment. The effect of RGal on macromolecular permeability was reduced with GFX pre‐treatment (500 nM) by 75.3% (n=5, p<0.01). However, the ability of GFX to block the RGal‐mediated increase in barrier function was not mimicked in monolayers pre‐treated with PKCz pseudosubstrate inhibitor (10 mM, n=6) or Go6976 (10 nM, n=5). (2) While RGal had no significant effect on tight junction gene expression 6h post‐treatment, it upregulated the expression of inflammatory response genes including neutrophil regulatory genes such as CXCL8. Consistent with gene expression data, multiplex protein assay revealed an upregulation in CXCL8 family chemokines and G‐CSF as early as 2h in cell lysates and 6h in the supernatant. Conclusions The RGal‐mediated increase in intestinal epithelial barrier function is dependent on PKC. Transcriptionally, RGal upregulates the expression of inflammatory mediators such as CXCL8. Understanding the mechanism of the RGal‐induced increase in intestinal epithelial barrier function may allow for leverage of these mechanisms to treat IBD. Support or Funding Information Project supported by the NSERC (Canada), J.S. supported by an UGREF from the American Physiological Society. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».