伊犁黑蜂蜂胶对不同生长状态白假丝酵母菌抑菌作用的体外研究

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(P0.05);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(P0.05)。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。

The inhibitory effects of propolis of Ili black bee on Candida albicans in vitro

Abstract: Objective To observe the effects of propolis of Ili black bee against Candida albicans in different states. Methods The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the propolis against Candida albicans in planktonic state were determined by using NCCLS M27-A2 yeast microdilution,combined with plate method.The biofilm model of Candida albicans was established in vitro; the minimal biofilm eradication concentration (MBEC) and the inhibitory effects were determined by using MTT method. Laser confocal microscopy (CLSM) was used to observe the inhibitory effects of different concentrations of propolis alcohol extracts on Candida albicans biofilm. Red and blue fluorescence staining was used for quantitative analysis.Results The MIC and MBC of the propolis of Ili black bee against planktonic Candida albicans was 4 mg/mL and was 8 mg/mL, respectively. The MBEC was 16 mg/mL. Along with the increase of the concentration of propolis, the inhibitory effect increased gradually, and the differences were statistically significant (P<0.05). CLSM revealed that the propolis of Ili black bee against Candida albicans significantly reduced the percentage of viable bacteria in biofilm and the biofilm activity, and the mortality rate of the fungi in biofilm showed an increasing trend along with the increasing of propolis concentration.Compared with negative control group, the difference was statistically significant (P<0.05).Conclusion The propolis of Ili black bee can significantly inhibit Candida albicans in different states in vitro.