白色念珠菌生物被膜体外模型的建立及分析

发布时间：2018-04-24 09:35

本文选题：白色念珠菌 + 生物被膜　；参考：《重庆医科大学》2010年硕士论文

【摘要】： 第一部分【目的】建立白色念珠菌(Candida albicans)生物被膜(Biofilm,BF)体外模型,运用多种形态学方法检测,初步探讨白色念珠菌BF相关感染发生机制。【方法】体外建立2、8、24,48及72 h时间组白色念珠菌BF模型,通过银染、荧光探针FITC-conA和SYTO9/PI标记等不同途径,结合荧光显微镜、扫描电镜(scanning electron microscope, SEM),激光共聚焦显微镜(Confocal Laser Scanning Microscopy,CLSM)观察不同时期白色念珠菌BF形成情况。【结果】白色念珠菌能够在玻片上形成典型的BF,通过荧光显微镜,SEM和CLSM完成对白色念珠菌BF形成过程动态观察,白色念珠菌随着时间增加逐渐增加,48 h形成初步生物被膜,72 h结构更加复杂和成熟。【结论】平板法能够较好地建立白色念珠菌BF体外模型,并可利用多种方式检测。第二部分【目的】应用ISA软件及Image-Pro Plus 6.0软件定量化分析白色念珠菌BF形成发展过程中变化,为进一步深研究其耐药机制及干预提供基础。【方法】将结合荧光探针SYTO9/PI标记的CLSM图片分别导入ISA软件及Image-Pro Plus 6.0软件分析获得白色念珠菌BF相关结构参数定量化数据。【结果】①ISA软件定量化分析显示,SYTO9/PI荧光探针标记的生物被膜模型的区域孔径(AP)在24, 48和72 h分别为0.95±0.06, 0.89±0.01和0.83±0.01,平均扩散距离(ADD)分别为1.16±0.13, 1.26±0.06和2.43±0.76,结构熵(TE)分别为4.87±0.34, 5.18±0.35和5.47±0.16,两两比较,差异均有显著性(P0.05)。②Image-Pro Plus 6.0软件分析显示,生物被膜内真菌死亡率分别为(34.71±2.72)%, (36.63±4.20)%和( 47.41±2.53)%,与24 h及48 h相比,72 h真菌死亡率增加具有显著性(P0.05)。【结论】利用ISA软件及Image-Pro Plus 6.0软件定量化分析可以反映白色念珠菌BF形成过程中的空间结构及死活菌比例变化特征。
[Abstract]:Part one [objective] to establish Candida albicansa biofilm biofilm BFs model in vitro and to investigate the mechanism of BF- related infection of Candida albicansa using various morphological methods. [methods] the BF model of Candida albicans was established in vitro by silver staining, fluorescence probe FITC-conA and SYTO9/PI labeling. Scanning electron microscopy (SEM) and confocal Laser Scanning microscopy (CLSM) were used to observe the formation of Candida albicans in different periods. [results] Candida albicans were able to form typical BFs on glass slides. The formation process of BF was observed by fluorescence microscope and CLSM. The structure of Candida albicans was more complex and mature at 72 h after 48 h of initial biofilm formation. [conclusion] the in vitro model of Candida albicans BF can be established by plate method, and it can be detected in many ways. Part two [objective] to analyze quantitatively the changes in the formation and development of Candida albicans BF by ISA software and Image-Pro Plus 6.0 software, so as to provide a basis for further study on the mechanism of drug resistance and intervention. [methods] the CLSM images labeled with fluorescent probe SYTO9/PI were imported into ISA software and Image-Pro Plus 6.0 software respectively to obtain quantitative data of BF related structure parameters of Candida albicans. [results] the quantitative analysis of 1ISA software showed that the APs of the biofilm model labeled with fluorescent probes of SYTO9 / Pi were 0.95 卤0.06,0.89 卤0.01,0.83 卤0.01 at 24,48 and 72 h, respectively, and the average diffusion distances were 1.16 卤0.13,1.26 卤0.06 and 2.43 卤0.76, respectively, and the structure entropy was 4.87 卤0.34, respectively, and the mean diffusion distance was 1.16 卤0.13,1.26 卤0.06 and 2.43 卤0.76, respectively, and the average diffusion distance was 1.16 卤0.13,1.26 卤0.06 and 2.43 卤0.76, respectively. 5.18 卤0.35 and 5.47 卤0.16, respectively. The results showed that the mortality rates of biofilm fungi were 34.71 卤2.72%, 36.63 卤4.20% and (47.41 卤2.53)%, respectively. The mortality of fungi in biofilm was significantly higher than that in 24 h and 48 h. [conclusion] the quantitative analysis of ISA and Image-Pro Plus 6.0 software can reflect the spatial structure of Candida albicans during the formation of BF and the variation characteristics of the proportion of live and dead bacteria.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R379.4

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