氟尼辛葡甲胺对克柔念珠菌生物膜形成的影响及联合唑类药物效果评价

摘要/Abstract

摘要：

目的本研究旨在评价氟尼辛葡甲胺（FM）对克柔念珠菌生物膜形成的抑制作用以及和唑类药物联用对克柔念珠菌的体外抗菌作用。方法以患真菌性乳房炎奶牛奶样临床分离克柔念珠菌为试验菌株，采用微量稀释法测定最小抑菌浓度（MIC）、最小杀菌浓度（MFC）初步探讨FM对克柔念珠菌的抑菌活性。水-烃两相法测定细胞表面疏水性（CSH）并通过XTT法测定FM对克柔念珠菌生物膜形成早期1~3 h黏附阶段的影响以及培养24 h后成熟生物膜的影响；以部分抑菌浓度指数（FICI）作为检测指标，通过棋盘法检测与唑类药物联用对克柔念珠菌体外静态抗菌作用；采用酶标比浊法测定绘制不同药物处理后的时间-抑菌曲线。结果 FM单药对克柔念珠菌的MIC均为0.625 mg/mL；FM可以降低CSH，对不同生长阶段（早期黏附、成熟）的克柔念珠菌生物膜均有显著抑制作用，抑制作用具有时间和浓度依赖性；联合用药发现，FM与伊曲康唑联用抗克柔念珠菌的FICI介于0.5~1.5之间，表现为协同/相加作用；与酮康唑联合用药的FICI 指数介于0.375~1.25之间，表现为协同/相加作用；与氟康唑联用则表现为无关；时间-抑菌曲线显示，部分唑类药物与FM联用能增强其对克柔念珠菌抑制作用，并维持较长的作用时间。结论 FM是一种有效的抗真菌增效剂，可以促进伊曲康唑和酮康唑的抗真菌活性。

关键词: 克柔念珠菌, 生物膜, 联合用药

Abstract:

Objective This study was designed to evaluate the antifungal activity of flunixin meglumine (FM) against Candida krusei biofilm and the antifungal effects of azole combined with flunixin meglumine (FM) in vitro. Method Candida krusei clinically isolated from cows with fungal mastitis was used as a test strain. The antifungal activity of FM against Candida krusei was preliminarily investigated by measuring the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) by microdilution method. The effects of FM on the early 1-3h adhesion phase of Candida krusei biofilm formation and the mature biofilm after 24h were determined by XTT method and effects of cell surface hydrophobicity (CSH) were determined by Water-hydrocarbon two-phase. The static antifungal activity of FM combined with antifungal azoles against Candida krusei in vitro was detected by checkerboard method using the partial FICI as the detection index. Enzyme-labeled turbidimetry was used to plot the time-kill curves of different drugs after treatment. Results The MIC of FM single drug against Candida krusei was 0.625 mg/mL; FM could reduce cell surface hydrophobicity,FM had significant inhibitory effect on Candida krusei biofilm at different growth stages (early adhesion and maturation), and its inhibitory effect was time-and concentration-dependent. The FICI of FM combined with itraconazole against Candida krusei ranged from 0.5 to 1.5, showing synergistic/additive effects. The FICI index of FM combined with ketoconazole was ranged from 0.375 to 1.25, showing a synergistic/additive effect. In the time-kill curves, the combination of FM and azoles could enhance the inhibitory effect of azoles on Candida krusei and maintain a longer action time. Conclusion FM is an effective antifungal synergist and can promote the antifungal activity of itraconazole and ketoconazole.

Key words: Candida krusei, biofilm, combination therapy

中图分类号:

R379.4

马生妍, 周学章. 氟尼辛葡甲胺对克柔念珠菌生物膜形成的影响及联合唑类药物效果评价[J]. 中国真菌学杂志, 2020, 15(1): 31-36.

MA Sheng-yan, ZHOU Xue-zhang. Study on the effect of flunixin meglumine on the biofilm formation of Candida krusei and on the combination with azoles[J]. Chinese Journal of Mycology, 2020, 15(1): 31-36.

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