Restorative effects of Acetobacter ghanensis on the pathogenicity of gliadin-induced modulation of tight junction-associated gene expression in intestinal epithelial cells

Abstract

Background/Aim: At present, a gluten-free diet is the only efficient way to treat celiac disease (CD). The development of novel approaches to lessen or counteract the pathogenic effects of gluten remains crucial for the treatment of CD. The aim in this investigation was to examine the restorative effects of Acetobacter ghanensis as a novel probiotic against gliadin-induced modulation in the barrier integrity of an intestinal epithelial cell (IEC) model (Caco-2). Methods: Fully differentiated Caco-2 cell monolayers were subjected to enzymatically digested gliadin with a pepsin and trypsin (PT) in the presence or absence of A. ghanensis for 90 min. The relative amounts of zonulin, zonula occludens-1 (ZO-1), claudin-1, and occludin mRNA expression were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Transepithelial electrical resistance (TEER) was evaluated to monitor the barrier integrity of cell monolayers. Statistical analyses were carried out using one- or two-way ANOVA followed by Tukey’s post-hoc analysis for multiple pairwise comparisons. Results: A significant upregulation (4.7-fold) of zonulin was noted in the PT-gliadin treated Caco-2 cells in comparison with the untreated controls (P<0.001). Conversely, gliadin-induced zonulin expression was markedly downregulated in the Caco-2 cells following exposure to A. ghanensis in the presence of PT-gliadin (P<0.001). Furthermore, prominent decreases in the mRNA expression levels of ZO-1 (45%) and occludin (40%) were seen in the PT-gliadin exposed Caco-2 cells compared to the untreated control cells (P<0.001). PT-gliadin in the Caco-2 cells did not significantly alter the mRNA levels of claudin-1 (P=0.172). Similarly to zonulin expression, the decreasing effect of PT-gliadin on ZO-1 was completely attenuated in the PT-gliadin-administrated Caco-2 cells following exposure to A. ghanensis (P<0.001). Conclusion: A. ghanensis restored the pathogenicity of PT-gliadin on intestinal barrier integrity.