不同时间点两性霉素B联合氟康唑抗真菌效果研究

Study on the antifungal effect of amphotericin B combined with fluconazole at different time points

目的探讨两性霉素B(amphotericin B,AmB)与氟康唑(fluconazole,FLC)不同时间点体外相互作用效果,为临床应用多烯类与唑类联合治疗计划提供参考。方法采用白色念珠菌标准株SC5314进行研究,以微量肉汤两倍稀释法测定抗真菌药物最小抑菌浓度(minimum inhibitory concentration,MIC)。在观察低浓度AmB对FLC抗真菌效果影响的实验组中,实验组为分别在不同时间点(0、2、4、6 h)向FLC中加入低浓度AmB(0.25μg/mL或0.125μg/mL),并设置在不同时间点向FLC中加入等量溶媒DMSO为FLC单药对照组;在观察低浓度FLC对AmB抗真菌效果影响的实验组中,实验组为分别在不同时间点(0、2、4、6 h)向AmB中加入低浓度FLC(0.06μg/mL或0.03μg/mL),并设置在不同时间点向AmB中加入等量溶媒DMSO为AmB单药对照组;设置溶媒DMSO对照组,分别在不同时间点加入等量溶媒DMSO及低浓度药物。各溶媒对照组、单药对照组及实验组分别观察复苏后菌落生长情况以评价药物浓度与时间的交互作用。结果FLC抑菌浓度(0.25μg/mL)处理下,加入低浓度AmB不影响FLC抗真菌效果,菌落计数差异倍数无明显变化(P>0.05)。AmB处理白色念珠菌后不同时间点加入低浓度FLC,与AmB单药对照组相比,0 h时菌落计数无明显变化(F=0.27,P=0.775);2~4 h真菌菌落计数达对照组1.74~1.93倍(P<0.001);6 h后,菌落计数较对照组无显著差异(P>0.05)。结论AmB与FLC相互作用具时间依赖性,联合用药早期FLC与AmB无相互作用,2~4 h低浓度FLC可减弱AmB杀菌效果,6 h以后不影响AmB抗真菌性能。

Objective To investigate the in vitro interaction of amphotericin B (AmB) and fluconazole (FLC) at dif⁃ferent time points and provide a reference for clinical combined treatment therapy of polyenes and azoles. MethodsCandida albicans ATCC SC5314 was used in the study. The minimum inhibitory concentration (MIC) of antifungaldrugs was determined using the double microdilution broth method. The same amount of DMSO and low concentrationdrugs were added to the DMSO treatment group at different time points (0, 2, 4, 6 h) to determine whether the solvent background environment affected the growth of Candida albicans. In the experimental group, to observe the effect of lowconcentration AmB on the antifungal effect of FLC, the experimental group was administered a low concentration ofAmB (0.25 μg/mL or 0.125 μg/mL) added to FLC at different time points (0, 2, 4, 6 h), and the same amount of DMSOwas added to FLC at different time points in the single drug control group. In the experimental group, to observe the ef⁃fect of low concentration of FLC on the antifungal effect of AmB, the experimental group was administered a low concen⁃tration of FLC (0.06 μg/mL or 0.03 μg/mL) in AmB at different time points (0, 2, 4, 6 h), and the same amount ofDMSO was used at different time points as the single drug control group. In the solvent group, the same amounts ofDMSO and low concentration drugs were added at different time points. After resuscitation, the colony growth of eachsolvent control group, single⁃drug control group and experimental group was observed to evaluate the interaction be⁃tween drug concentration and time. Compared with the AmB single⁃drug control group, there was no significant changein the experimental group with added low concentrations of FLC at 0 h (F=0.27, P=0.775), which was 1.74⁃1.93times that of the control group at 2⁃4 h (P < 0.001), and there was no significant difference in colony count after 6 h(P > 0.05). Results Under the treatment of FLC at an inhibitory concentration (0.25 μg/ml), adding low concentrationAMB did not affect the antifungal effect of FLC, and the multiple of colony count differences were not significant (P >0.05). Conclusion The interaction between AmB and FLC was time⁃dependent. At the early stage (0 h), the interac⁃tion effect between fluconazole and amphotericin B was not clear. The fungicidal effect of AmB could be weakenedwhen FLC was supplied at 2⁃4 h, and the effect of FLC on AmB was absent after 6 h.