BDSF抑制白念珠菌生物膜的形成

发布时间：2018-03-24 00:08

本文选题：白念珠菌　切入点：生物膜　出处：《复旦大学》2011年硕士论文

【摘要】：白念珠菌是一种极其重要的条件致病真菌,白念珠菌可形成生物膜,极大的增强了其抗药性,是临床上的一大难题。本实验中通过显微镜观察的方法,检测了cis-BDSF和trans-BDSF在体外对于白念珠菌的生长、出芽以及生物膜形成的影响。通过静置微孔板法构建白念珠菌生物膜模型,利用XTT法定量测定在白念珠菌生物膜生长的不同时间加入不同浓度的cis-BDSF和trans-BDSF对白念珠菌生物膜形成的抑制作用,并且通过显微镜观察其对生物膜结构和形成的影响,northern杂交检测cis-BDSF和trans-BDSF处理过的生物膜中菌丝特异性基因HWP1的表达变化。结果显示300μM的cis-BDSF和trans-BDSF均可抑制白念珠菌酵母相的生长,出芽抑制率可达70%。在0小时和1小时时加入300μM的cis-BDSF和trans-BDSF,对于生物膜的抑制率均可达90%(0小时)和60%(1小时),显著高于法尼醇和DSF。然而,四种药均不能对已经形成的生物膜造成影响。Northern杂交结果表明cis-BDSF和trans-BDSF可下调HWP1的表达。实时荧光定量分析同样显示经60μM的cis-BDSF和trans-BDSF处理,白念珠菌生物膜模型中HWP1和ALS3的表达量分别下调约90%(cis-BDSF)和70-80%(trans-BDSF),证明在生物膜形成早期(0或1小时)加入cis-BDSF和trans-BDSF可以显著抑制白念珠菌生物膜的形成,低浓度(3μM、30μM和60μM)时主要通过抑制其形态转变及菌丝相的附着实现的,而高浓度(300μM)时则通过抑制形态转变和菌丝相附着以及酵母相的生长的联合作用实现的。Cis-BDSF和trans-BDSF作为可能的治疗方法值得进行进一步研究。
[Abstract]:Candida albicans is an extremely important condition pathogenic fungus, Candida albicans can form biofilm, greatly enhance its resistance, which is a difficult problem in clinic. The effects of cis-BDSF and trans-BDSF on the growth, budding and biofilm formation of Candida albicans in vitro were investigated. The inhibition of Candida albicans biofilm formation by adding different concentrations of cis-BDSF and trans-BDSF at different time of biofilm growth was measured by XTT assay. The effects on the structure and formation of biofilm were observed by microscope. The changes of HWP1 expression of mycelium specific gene in biofilm treated with cis-BDSF and trans-BDSF were detected by Northern hybridization. The results showed that 300 渭 M of cis-BDSF and trans-BDSF could inhibit the growth of Candida albicans yeast phase. At 0 and 1 hour, the inhibition rates of cis-BDSF and trans-BDSFs on biofilm were up to 90 h and 60% respectively, which were significantly higher than those of Fannitol and DSF. Northern blot analysis showed that cis-BDSF and trans-BDSF could down-regulate the expression of HWP1. Real-time fluorescence quantitative analysis showed that they were also treated with 60 渭 M cis-BDSF and trans-BDSF. In Candida albicans biofilm model, the expression of HWP1 and ALS3 were down-regulated by 90 cis-BDSFs and 70-80BDSFs, respectively. The results showed that cis-BDSF and trans-BDSF could significantly inhibit the formation of Candida albicans biofilm in the early stage of biofilm formation. At low concentrations of 3 渭 M, 30 渭 M and 60 渭 M, it was mainly achieved by inhibiting the morphological transformation and the attachment of hyphae phase. The possible therapeutic methods of Cis-BDSF and trans-BDSF by inhibiting morphological transformation, hyphal attachment and yeast growth at high concentration (300 渭 M) are worth further study.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R379

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