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. 2016 May 9;7:120. doi: 10.3389/fphar.2016.00120

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Copyright © 2016 Esposito, Nobile, Gigli, Seguella, Pesce, d’Alessandro, Bruzzese, Capoccia, Steardo, Cuomo and Sarnelli.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effects of increasing concentrations of rifaximin (0.1, 1.0, and 10 μM) alone and rifaximin plus ketoconazole (10 μM) against TcdA (30 ng/ml) in Caco-2 cells: (A) 24-h time course TEER changes (n = 4); (B) MTT cell viability absorbance at 24 h (n = 5); (C) Immunoreactive bands corresponding to Bax, ZO-1, and occludin expression at 24 h following the TcdA challenge; (D) Relative densitometric analysis of immunoreactive bands (arbitrary units normalized against the expression of the housekeeping GAPDH protein; n = 5), and (E) Immunofluorescent staining showing the effects of TcdA challenge on ZO-1 and occludin co-expression at 24 h. Nuclei were also investigated using DAPI staining (Scale bar = 25 μm). Results are expressed as mean ± SEM of experiments performed in triplicate. ^∗∗∗p < 0.001 and ^∗∗p < 0.01 vs. vehicle group; ^∘∘∘p < 0.001, ^∘∘p < 0.01 and °p < 0.05 vs. TcdA group.