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  本文選題：白念珠菌　切入點：生物膜　出處：《復(fù)旦大學(xué)》2011年碩士論文

【摘要】：白念珠菌是一種極其重要的條件致病真菌,白念珠菌可形成生物膜,極大的增強了其抗藥性,是臨床上的一大難題。本實驗中通過顯微鏡觀察的方法,檢測了cis-BDSF和trans-BDSF在體外對于白念珠菌的生長、出芽以及生物膜形成的影響。通過靜置微孔板法構(gòu)建白念珠菌生物膜模型,利用XTT法定量測定在白念珠菌生物膜生長的不同時間加入不同濃度的cis-BDSF和trans-BDSF對白念珠菌生物膜形成的抑制作用,并且通過顯微鏡觀察其對生物膜結(jié)構(gòu)和形成的影響,northern雜交檢測cis-BDSF和trans-BDSF處理過的生物膜中菌絲特異性基因HWP1的表達變化。結(jié)果顯示300μM的cis-BDSF和trans-BDSF均可抑制白念珠菌酵母相的生長,出芽抑制率可達70%。在0小時和1小時時加入300μM的cis-BDSF和trans-BDSF,對于生物膜的抑制率均可達90%(0小時)和60%(1小時),顯著高于法尼醇和DSF。然而,四種藥均不能對已經(jīng)形成的生物膜造成影響。Northern雜交結(jié)果表明cis-BDSF和trans-BDSF可下調(diào)HWP1的表達。實時熒光定量分析同樣顯示經(jīng)60μM的cis-BDSF和trans-BDSF處理,白念珠菌生物膜模型中HWP1和ALS3的表達量分別下調(diào)約90%(cis-BDSF)和70-80%(trans-BDSF),證明在生物膜形成早期(0或1小時)加入cis-BDSF和trans-BDSF可以顯著抑制白念珠菌生物膜的形成,低濃度(3μM、30μM和60μM)時主要通過抑制其形態(tài)轉(zhuǎn)變及菌絲相的附著實現(xiàn)的,而高濃度(300μM)時則通過抑制形態(tài)轉(zhuǎn)變和菌絲相附著以及酵母相的生長的聯(lián)合作用實現(xiàn)的。Cis-BDSF和trans-BDSF作為可能的治療方法值得進行進一步研究。
[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.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R379

【參考文獻】

相關(guān)期刊論文 前8條

1 石宏宴;馬榮;王彥;;149株念株菌鑒定與藥敏分析[J];大連醫(yī)科大學(xué)學(xué)報;2007年04期

2 劉小平,樊尚榮,李建武;婦科門診患者外陰陰道念珠菌病的檢出率和抗真菌藥物敏感性研究[J];中國婦產(chǎn)科臨床雜志;2004年02期

3 羅利龍;婁瑞娟;邱健;宋水山;;細菌群體感應(yīng)及其干擾策略的研究進展[J];生物學(xué)雜志;2010年06期

4 沈亮亮;葉元康;;氟康唑與5-氟胞嘧啶體外聯(lián)合抗白念珠菌臨床分離株的研究[J];同濟大學(xué)學(xué)報(醫(yī)學(xué)版);2005年06期

5 王丹敏;靳穎;董小青;劉麗英;;白色念珠菌生物膜生長動力學(xué)分析和形態(tài)學(xué)觀察[J];武警醫(yī)學(xué)院學(xué)報;2010年09期

6 苑天紅,王明永,吳升偉,王正蓉,吳承龍;白念珠菌二相性與致病性關(guān)系[J];中國公共衛(wèi)生;2005年01期

7 王丹敏,韓景田,董小青,屈野;白色念珠菌蛋白酶與其毒力關(guān)系的研究[J];中國微生態(tài)學(xué)雜志;2000年04期

8 溫旺榮,王德春,陳紅,朱忠勇;白念珠菌的毒力研究——蛋白酶活力的測定[J];中國人獸共患病雜志;1999年06期

，

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