Table 1.
Gene-specific primers used for RealTime-PCR.
Fig 1.
Three-dimensional plots of fluconazole combined with tested calcium channel blockers against CA10 (resistant) using the MATLAB program.
The ΔE values obtained for each combination can be depicted on the z axis to construct a 3D surface plot. Peaks above and below the 0 plane indicate synergistic and antagonistic combinations, respectively. The 0 plane indicates the absence of SS interaction. The color-coding on the right indicates that the closer to red in the top of the bar, the more effective the drug combinations.
Table 2.
The combined antifungal effects of fluconazole with calcium channel blockers against resistant Candida albicans evaluated by FICI.
Fig 2.
The Time-kill curves of fluconazole in combination with tested calcium channel blockers against resistant Candida albicans (CA10).
Cells were diluted in RPMI 1640 medium containing FLC, AML, NIF, BEN, FNZ and combination of FLC with the four CCBs. The concentration of FLC was 1 μg ml-1 when combined with AML (8 μg ml-1) and FNZ (16 μg ml-1), and the concentration of FLC was 2 μg ml-1 when combined with NIF (8 μg ml-1) and BEN (16 μg ml-1). Values represent the means ±standard deviation of three replicates. * P < 0.01 and at least 2.0-fold compared to the fluconazole group.
Fig 3.
Relative expression of CNA1, CNB1 and YVC1 following treatment with fluconazole (FLC) and amlodipine (AML) alone or in combination in CA10.
Cells were treated with fluconazole at 1 μg ml-1, amlodipine at 16 μg ml-1 alone or in combination. Total RNA was extracted and reversely transcribed to cDNA. cDNA was then used for real-time quantitative PCR to detect expression levels of CNA1, CNB1 and YVC1. Values represent the means ± standard deviation of three replicates. * P < 0.01 and at least 2.0-fold compared to the control group.
Fig 4.
The influence of tested calcium channel blockers on fluconazole efflux.
(A). Relative expression of CDR1, CDR1 and MDR1 following treatment with fluconazole (FLC) and amlodipine (AML) alone or in combination in CA10. Cells were treated with fluconazole at 1 μg ml-1, amlodipine at 16 μg ml-1 alone or in combination. Total RNA was extracted and reversely transcribed to cDNA. cDNA was then used for real-time quantitative PCR to detect expression levels of CDR1, CDR1 and MDR1. Values represent the means ± standard deviation of three replicates. (B). The influence of amlodipine, nifedipine, benifdipine and flunarizine on efflux of fluconazole was tested by rhodamine 6G assay. There was almost no difference in fluorescence intensity with or without the tested calcium channel blocker. Values represent the means ± standard deviation of three replicates. * P < 0.01 and at least 2.0-fold compared to the control group.