口腔疾病防治 ›› 2021, Vol. 29 ›› Issue (1): 20-26.DOI: 10.12016/j.issn.2096-1456.2021.01.003

• 基础研究 • 上一篇    下一篇

不同时间点两性霉素B联合氟康唑抗真菌效果研究

朱乘光1(),叶星辰2,任彪2,周学东1,程磊1()   

  1. 1.口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心 四川大学华西口腔医院牙体牙髓病科,四川 成都(610041)
    2.口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心,四川 成都(610041)
  • 收稿日期:2020-01-22 修回日期:2020-08-06 出版日期:2021-01-20 发布日期:2020-11-18
  • 通讯作者: 程磊
  • 作者简介:朱乘光,博士研究生,Email:253434077@qq.com
  • 基金资助:
    国家自然科学基金面上项目(81870778)

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

ZHU Chengguang1(),YE Xingchen2,REN Biao2,ZHOU Xuedong1,CHENG Lei1()   

  1. 1. Department of Operative Dentistry and Endodontics, West China Hospital of Stomatology, State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Sichuan University, Chengdu 610041, China
    2. State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Chengdu 610041, China
  • Received:2020-01-22 Revised:2020-08-06 Online:2021-01-20 Published:2020-11-18
  • Contact: Lei CHENG
  • Supported by:
    National Natural Science Foundation of China(81870778)

摘要:

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(0.25 μg/mL或0.125 μg/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(0.06 μg/mL或0.03 μg/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(0.25 μg/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。

关键词: 白色念珠菌, 两性霉素B, 氟康唑, 体外抗真菌活性, 最小抑菌浓度, 联合用药, 时间依赖性

Abstract:

Objective To investigate the in vitro interaction of amphotericin B (AmB) and fluconazole (FLC) at different time points and provide a reference for clinical combined treatment therapy of polyenes and azoles. Methods Candida albicans ATCC SC5314 was used in the study. The minimum inhibitory concentration (MIC) of antifungal drugs was determined using the double microdilution broth method. The same amount of DMSO and low concentration drugs 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 low concentration AmB on the antifungal effect of FLC, the experimental group was administered a low concentration of AmB (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 DMSO was added to FLC at different time points in the single drug control group. In the experimental group, to observe the effect of low concentration of FLC on the antifungal effect of AmB, the experimental group was administered a low concentration 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 of DMSO was used at different time points as the single drug control group. In the solvent group, the same amounts of DMSO and low concentration drugs were added at different time points. After resuscitation, the colony growth of each solvent control group, single-drug control group and experimental group was observed to evaluate the interaction between drug concentration and time. Compared with the AmB single-drug control group, there was no significant change in the experimental group with added low concentrations of FLC at 0 h (F=0.27, P=0.775), which was 1.74-1.93 times 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 concentration AMB 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 interaction effect between fluconazole and amphotericin B was not clear. The fungicidal effect of AmB could be weakened when FLC was supplied at 2-4 h, and the effect of FLC on AmB was absent after 6 h.

Key words: Candida albicans, amphotericin B, fluconazole, in vitro antifungal activity, minimal inhibitory concentration, drug combination, time-dependence

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