Rate of amiloride-sensitive Na+ uptake into cultured rumen epithelial cells was studied in order to clarify the influence of culture conditions on Na+/H+ exchange (NHE). Cell cultures were exposed to Na-n-butyrate or not for seven days or subcultured. On the 14th day of culturing, primary cell cultures without butyrate exposure showed both non-stratified and stratified growth. Na-n-butyrate treated 14-day-old cultures and 3-day-old subcultures contained mostly non-stratified, i.e. non-keratinised cells. Both n-butyrate treatment and subculturing increased total and amiloride-sensitive Na+ uptake. Our results indicate that Na+ uptake via NHE is determined by the amount and the ratio of non-stratified (non-keratinised) cells.
Authors:K. Szekér, E. Németh, Sz. Kun, J. Beczner and P. Gálfi
DSM 20017 and
B3.2 to Caco-2 cell line was investigated in vitro. The adhesion ability of the tested strains was quantified with three methods: fluorescent-labelling, Gram-staining — followed by cell counting and image analysis — and plate count enumeration in order to compare the different detection methods. Results were in good correlation in terms of number of adhered bacteria, however, aggregate formation resulted in a significantly lower result with plate count enumeration in case of
2750. Percent coverage was found to be an appropriate method to compare adhesion ability of the strains, provided the cell sizes are similar. Gram-staining gives satisfactory results, however, fluorescent staining was not a suitable method in this study, since fluorescent dye hexidium iodide also labelled the intestinal cells.
Authors:K. Szekér, J. Beczner, A. Halász, Á. Mayer, J.M. Rezessy-Szabó and P. Gálfi
The adhesion of twenty-six Lactobacillusstrains to two intestinal cell lines (Caco-2P and IEC-18) and 21 Bifidobacteriumstrains to Caco-2P cells was investigated. Non-specific adherence was determined on the surface of culture plates. The effect of short chain fatty acids (SCFA) on epithelial cells, and bacterial adhesion were investigated by Na-n-butyrate treatment. The adherence of LAB and bifidobacteria greatly varied in a strain-dependent manner. The adherence of LAB was better to IEC-18 cells than to Caco-2P cells, and bifidobacteria adhered better to Caco-2P cells than the LAB. Some strains adhered well or even better to the background than to the cells, which queries the specificity of adhesion of these strains. Na-n-butyrate treatment stimulated the differentiation of IEC-18 cells and therefore increased the number of adherent bacteria, probably because only the cell surface increased not the number of epitopes.
Authors:E. Paszti-Gere, E. Csibrik-Nemeth, K. Szeker, R. Csizinszky, O. Palocz, O. Farkas and P. Galfi
Recently, there has been a growing interest to replace antibiotics’ administration with the application of probiotics. The aim of our investigations was to reveal the influence of spent culture supernatant of Lactobacillus plantarum 2142 on the response of enterocytes to oxidative stress, and the spent culture supernatant’s ability to protect them from oxidative injury. The experiments were performed on non-carcinogenic porcine epithelial cell line, IPEC-J2 isolated from a neonatal piglet and on human colon adenocarcinoma cell line, Caco-2. The cells cultured on membrane inserts were treated with millimolar hydrogen peroxide solution to provoke oxidative stress. The peroxide-triggered cell response profile was evaluated via determination of change in transepithelial electrical resistance, quantification of extent of cell death by 4’,6-diamidino-2 phenylindole (DAPI) staining and via estimation of proinflammatory cytokine, IL-8 production using ELISA technique. Non-starter lactobacilli supernatant-mediated inhibition of peroxide-triggered upregulation of IL-8 production confirmed the antiinflammatory properties of active metabolites produced by Lactobacillus plantarum 2142 in acute oxidative stress.