Increasing the Fungicidal Action of Amphotericin B by Inhibiting the Nitric Oxide-Dependent Tolerance Pathway
AmB induced accumulation of superoxide and nitric oxide radicals in C. albicans and decreased the number of cells that are able to proliferate. Exponential C. albicans cultures were treated with different concentrations of AmB for 3 h at room temperature and subjected to flow cytometry or plating assays. (a) Levels of superoxide radical detected by dihydroethidium (DHE) fluorescence and flow cytometry. (b) Levels of nitric oxide radical detected by 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM DA) fluorescence and flow cytometry. (c) Number of CFU/mL in Log-scale, assessed by plating assays and CFU counting. Means and standard errors of the means (SEMs) of at least 3 independent biological experiments () are presented. Two-way ANOVA followed by Dunnett multiple comparison test was performed to analyse statistically significant differences in the number of DHE- and DAF-FM DA-positive cells and cells able to proliferate between control treatment and treatment with different concentrations of AmB. and represent and , respectively.