In order to evaluate the release of zonulin during the time of ob

In order to evaluate the release of zonulin during the time of observation, the area under the curve (AUC) was calculated. All data are expressed as mean and SEM. Differences were considered significant at P < 0.05. A specific software package (SigmaStat for Windows version 3.00 SPSS Inc. San Jose, CA, USA) was used. Results Effects of gliadin and L.GG

treatments on Caco-2 monolayer barrier function (TER and lactulose flux) TER measurements were determined after the addition of viable L.GG, L.GG-HK and L.GG-CM to polarized monolayers of Caco-2 cells seeded on Transwell filter inserts. TER was measured before the addition of bacteria, at time zero (immediately after the bacteria administration) and then at various time intervals ranging from

30 min to 6 h. A slight and not significant Selleck CHIR 99021 increase in TER was observed after 90 min from the addition of viable L.GG, as well as L.GG-HK and L.GG-CM to Caco-2 cells compared to control cells (data not shown). Addition of gliadin to the mucosal side of Caco-2 monolayers led to an immediate lowering in TER (Figure 1). The TER of gliadin-treated Caco-2 cells immediately after selleck kinase inhibitor the gliadin administration (time zero) Torin 2 mouse decreased to 30% of TER measured before treatment and started to recover after 90 min of incubation. The co-administration of viable L.GG, L.GG-HK and L.GG-CM with gliadin had a significant (P < 0.05) reversible effect on the recovery of TER starting 60 min post-incubation compared to gliadin-treated cells. After 6 h, the reversion of TER of viable L.GG, L.GG-HK and L.GG-CM to gliadin-treated cells reached 90%, 76% and 80% of their initial values before the addition of gliadin. Digestive enzyme Figure 1 Effects of supplementation of viable L.GG (10 8   CFU/ml), L.GG-HK

and L.GG-CM on gliadin-induced (1 mg/ml) TER decrease. All data represent the results of three different experiments (mean ± SEM). For each time of treatment, data were analyzed by Kruskal-Wallis analysis of variance and Dunn’s Multiple Comparison Test. (*) P < 0.05 compared to gliadin treated cells. To confirm that TER reduction involved the opening of intercellular TJs, the mucosal to serosal transport of the paracellular marker lactulose was also monitored. No effect on lactulose flux was observed after 90 min from the addition of viable L.GG, as well as L.GG-HK and L.GG-CM to Caco-2 cells compared to control cells (data not shown). By opposite, in monolayers treated with gliadin, a significant increase (P < 0.05) in serosal lactulose (0.077 ± 0.04 μg/ml) was observed 90 min after gliadin exposure compared to untreated monolayers (0.025 ± 0.02 μg/ml). The co-administration of viable L.GG, L.GG-HK and L.GG-CM antagonized the increased paracellular lactulose transport due to gliadin treatment (viable L.GG: 0.03 ± 0.02 μg/ml; L.GG-HK: 0.039 ± 0.01 μg/ml; L.GG-CM: 0.04 ± 0.01 μg/ml). Effects of gliadin and L.GG treatments on zonulin release Viable L.GG, L.GG-HK and L.

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