伊犁黑蜂蜂胶对不同状态白色念珠菌作用的实验研究

发布时间：2018-05-10 12:46

本文选题：黑蜂蜂胶 + 白色念珠菌　；参考：《新疆医科大学》2017年硕士论文

【摘要】：目的:研究新疆伊犁黑蜂蜂胶对白色念珠菌的抑菌作用,通过蜂胶的抑菌和杀菌效果,进一步研究游离状态下白色念珠菌的生长、产酸耐酸、粘附的抑制作用以及通过观察蜂胶对白色念珠菌生物膜活性的影响,探讨伊犁黑蜂蜂胶可能的防龋机制,为开拓新疆天然药物防龋及其新用途提供实验参考。方法:采用NCCLS M27-A2酵母菌微量稀释法结合平板法测定蜂胶对游离状态下白色念珠菌的最小抑菌浓度和最低杀菌浓度;通过测定最小抑菌浓度以下4个不同梯度浓度蜂胶培养基的前后OD540变化值和pH变化值,判断蜂胶对游离状态下白色念珠菌的产酸耐酸的影响;采用玻壁法计算黏附比探讨不同梯度浓度蜂胶抑制白色念珠菌的黏附情况;玻片法在体外构建白色念珠菌生物膜模型,采用MTT比色法检测蜂胶对白色念珠菌生物膜状态下的最低生物膜清除浓度,激光共聚焦显微镜观察不同浓度蜂胶醇提取物对同一时间白色念珠菌生物膜的抑菌效果,并进行红蓝荧光染色定量分析。结果:伊犁黑蜂蜂胶对白色念珠菌的最小抑菌浓度为4 mg/mL,最低杀菌浓度为8 mg/m L;蜂胶可使各游离状态下的白色念珠菌ΔpH降低,蜂胶各组与对照组相比差异均有统计学意义(P0.05);蜂胶可使各游离状态下的白色念珠菌在酸性环境中ΔOD540降低,蜂胶各组与对照组相比差异均有统计学意义(P0.05);蜂胶可使各游离状态下的白色念珠菌黏附比值降低,蜂胶各组与对照组相比差异均有统计学意义(P0.05)。伊犁黑蜂蜂胶对白色念珠菌的最低生物膜清除浓度是16 mg/mL,随着药物浓度的增加抑菌性逐渐增大;激光共聚焦显微镜观察,伊犁黑蜂蜂胶作用于白色念珠菌生物膜后可使生物膜内活菌比例下降,生物膜活性降低。结论:伊犁黑蜂蜂胶在体外对浮游状态和生物膜状态下白色念珠菌的生长均起到了明显的抑制作用,并对浮游菌的产酸、耐酸、粘附均起到了抑制作用。
[Abstract]:Objective: to study the bacteriostatic effect of Propolis on Candida albicans in Xinjiang, and to further study the growth of Candida albicans in free state. By observing the effect of propolis on the biofilm activity of Candida albicans, the possible anti-caries mechanism of propolis was discussed. Methods: the minimal inhibitory concentration and bactericidal concentration of propolis against Candida albicans in free state were determined by NCCLS M27-A2 microdilution method and plate method. The effects of propolis on the acid-resistance of Candida albicans in free condition were determined by measuring the changes of OD540 and pH before and after the four different gradient concentrations of propolis medium with the minimum inhibitory concentration of propolis. The adhesion ratio of Candida albicans was calculated by glass wall method, and the adhesion inhibition of Candida albicans by propolis with different gradient concentrations was studied, and the biofilm model of Candida albicans was established by glass slide method. The minimum biofilm scavenging concentration of propolis against Candida albicans biofilm was detected by MTT colorimetry. The inhibitory effect of propolis alcohol extract with different concentrations on Candida albicans biofilm at the same time was observed by confocal laser microscope. The quantitative analysis of red and blue fluorescence staining was carried out. Results: the minimum inhibitory concentration of propolis against Candida albicans was 4 mg / mL and the minimum bactericidal concentration was 8 mg/m / L. Propolis could reduce the 螖 pH of Candida albicans in different free states. Propolis could reduce the 螖 OD540 of Candida albicans in acidic environment. Propolis can reduce the adhesion ratio of Candida albicans in each free state, and there is a significant difference between the propolis groups and the control group (P 0.05). The minimum biofilm scavenging concentration of propolis against Candida albicans was 16 mg / mL, which increased with the increase of drug concentration. After acting on Candida albicans biofilm, the ratio of living bacteria in biofilm and the activity of biofilm were decreased. Conclusion: Ili bee propolis can inhibit the growth of Candida albicans in both planktonic state and biofilm state in vitro, and inhibit the acid production, acid resistance and adhesion of phytoplankton.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R781.1

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