Polyamines are in nmol/mg protein. The administration of gliadin selleck chemical to Caco-2 cells led to a significant increase (P < 0.05) in the spermidine (+35%), spermine (+42%) and total polyamine content (+46%) in comparison with untreated control cells. The supplementation of viable L.GG and L.GG-HK, but not L.GG-CM, on gliadin treated cells counteracted significantly (P < 0.05) the effects of gliadin on the polyamine profile. In particular, the contents in spermidine and spermine decreased
by 35.5% and 61.3%, respectively for viable L.GG. Overall, the percentage of reduction in the total polyamine content was by 50.7%. As concerns cells treated with gliadin and L.GG-HK, the reduction in spermidine and spermine content was equal to
23.6% and 19.8%, respectively. The total polyamine content was reduced by 23.9%. Effects of gliadin and L.GG treatments on ZO-1, Claudin-1 and Occludin expression To establish whether the changes in paracellular permeability on Caco-2 monolayers following gliadin and L.GG treatments were learn more associated with modifications in ZO-1, Claudin-1 and Occludin expression, mRNA and protein levels of the three proteins were quantified by qPCR and Western Blot analysis, respectively. When Caco-2 cells were exposed to viable L.GG, L.GG-HK and L.GG-CM for 6 h, a significant (P < 0.05) increase in the ZO-1, Claudin-1 and Occludin mRNA levels compared to control cells was observed only after viable bacteria treatment (Figure 3, panels A, B, and C). Figure 3 ZO-1, Claudin-1 and Occludin mRNA selleck chemicals llc levels in Caco-2 monolayers after 6 h of exposure to different probiotic and gliadin treatments. Panels A, B, and C report ZO-1, Claudin-1 and Occludin mRNA levels in Caco-2 monolayers after 6 h of exposure to viable L.GG (108 CFU/ml), heat killed L.GG (L.GG-HK) and L.GG conditioned medium (L.GG-CM). Data were analyzed by Kruskal-Wallis analysis of variance and Dunn’s Multiple Comparison Test. (*) P < 0.05 compared to control cells. Panels D, E and F report ZO-1, Claudin-1 and Occludin mRNA levels in Caco-2 monolayers after 6 h of exposure
to gliadin (1 mg/ml) alone or in combination with viable L.GG, L.GG-HK and L.GG-CM. Data were analyzed by Kruskal-Wallis analysis of variance and Dunn’s Multiple Comparison Test. Phosphoribosylglycinamide formyltransferase (*) P < 0.05 compared to gliadin treated cells. All data represent the results of three different experiments (mean ± SEM). The administration of gliadin did exert a slight and not significant down-regulatory effect on ZO-1 (−20.6%) and Occludin (−17.5%) expression, without affecting Claudin-1 one. By opposite, only the administration of viable L.GG in combination with gliadin caused a significant (P < 0.05) increase in the mRNA levels of all the tested proteins. In particular, ZO-1 and Claudin-1 increased more than tenfold and Occludin more than fourfold compared to gliadin-treated cells (Figure 3, panels D, E, and F). L.GG-HK and L.