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目的探究伊犁黑蜂蜂胶对白假丝酵母菌游离状态及其生物膜状态的影响。方法采用NCCLS M27-A2酵母微量稀释法结合平板法测定蜂胶对游离状态下白假丝酵母菌的最小抑菌浓度(MIC)和最低杀菌浓度(MBC);玻片法在体外构建白假丝酵母菌生物膜模型,采用MTT法检测蜂胶对白假丝酵母菌生物膜状态下的最低生物膜清除浓度(MBEC)及抑制作用;激光共聚焦显微镜(CLSM)观察不同浓度蜂胶醇提取物对同一时间白假丝酵母菌生物膜的抑菌效果,并进行红蓝荧光染色定量分析。结果伊犁黑蜂蜂胶对浮游白假丝酵母菌的MIC为4mg/mL,MBC为8mg/mL;伊犁黑蜂蜂胶对生物膜状态下白假丝酵母菌的MBEC为16mg/mL,随着药物浓度的增加抑菌性逐渐增大,差异具有统计学意义(P<0.05);CLSM观察,伊犁黑蜂蜂胶作用于白假丝酵母菌生物膜后可使生物膜内活菌比例降低,生物膜活性减弱,生物膜内实验菌死亡率随蜂胶浓度增加呈上升趋势,与阴性对照组比较差异有统计学意义(P<0.05)。结论伊犁黑蜂蜂胶在体外对不同状态下的白假丝酵母菌均起到了明显的抑制作用。
Objective To investigate the effect of Yili black bee propolis on the free state of Candida albicans and its biofilm status. Methods The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of propolis to Candida albicans in free state were determined by NCCLS M27-A2 yeast microdilution method and plate method. The Candida albicans Bacterial biofilm model was used to detect the minimum biofilm clearance (MBEC) and prophylactic effect of propolis on Candida albicans biofilm by MTT method. Laser confocal microscopy (CLSM) was used to observe the effects of different concentrations of propolis alcohol extract on the same time white Candida albicans biofilm antibacterial effect, and quantitative analysis of red and blue fluorescence staining. Results The MIC of Ili black bee propolis to P. albicans was 4 mg / mL and the MBC was 8 mg / mL. The MBEC of Yili black bee propolis to C. albicans in biofilm was 16 mg / mL, (P <0.05). The results of CLSM showed that the ratio of viable cells in biofilm decreased after treatment with Yili black bee propolis on the biofilm, and the biofilm activity The mortality of experimental bacteria in biofilm increased with the increasing of propolis concentration, which was significantly different from that of negative control group (P <0.05). Conclusion Ili black bee propolis in vitro in different states of Candida albicans have played a significant inhibitory effect.