中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (9): 641-644.

• 论著 • 上一篇    下一篇

白念珠菌磷脂酶表达水平与氟康唑耐药关系的实验研究

苏英1,李春阳2   

  1. 1. 山东大学齐鲁医院皮肤科
    2. 济南市山东大学齐鲁医院皮肤科
  • 收稿日期:2010-01-15 修回日期:2010-05-18 发布日期:2010-09-10
  • 通讯作者: 李春阳 E-mail:lichunyang@medmail.com.cn
  • 基金资助:

    白念珠菌多分子耐药机制的研究及ERG11基因点突变芯片的制备

Relationship between phospholipase expression and fluconazole resistance in Candida albicans

  • Received:2010-01-15 Revised:2010-05-18 Published:2010-09-10

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摘要：

目的 探讨白念珠菌毒力因子磷脂酶B1与耐药间的关系。方法 实验用白念珠菌氟康唑耐药株和敏感株各15株，提取白念珠菌总RNA，应用RT－PCR检测比较其耐药株与敏感株磷脂酶B1 mRNA的表达。盐析法提取白念珠菌耐药株与敏感株胞外蛋白，P0013B 放射免疫沉淀分析（RIPA）裂解液（强）裂解提取细胞内蛋白质，以β-肌动蛋白为内参照，用Western印迹法分别比较胞内和胞外耐药株和敏感株磷脂酶B1蛋白的表达水平。结果 耐药株和敏感株磷脂酶B1mRNA的相对表达量分别为0.6173 ± 0.1090和0.2653 ± 0.0935，耐药株磷脂酶B1mRNA的表达强于敏感株（t = 2.452，P < 0.05）。耐药株和敏感株胞内磷脂酶B1蛋白相对表达量为0.4145 ± 0.2773和0.1825 ± 0.1831，胞外磷脂酶B1蛋白相对表达量为0.2720 ± 0.2194和0.0688 ± 0.0631，耐药株胞内磷脂酶B1和细胞外分泌的磷脂酶B1蛋白表达水平均高于敏感株（t = 2.703、3.443，P值均 < 0.05）。 结论 白念珠菌耐药株毒力因子磷脂酶B1 mRNA及蛋白表达水平均高于敏感株。白念珠菌磷脂酶表达水平与耐药的发生可能存在一定关系。

关键词: 念珠菌, 白色, 溶血磷脂酶, 抗药性, 真菌

Abstract:

Objective To explore the relationship between antifungal resistance and expression of a virulence factor phospholipase B1 in Candida albicans. Methods Total RNA was extracted from 15 fluconazole-resistant and 15 fluconazole-sensitive Candida albicans isolates. RT-PCR was performed to detect the expression of phospholipase B1 mRNA. Extracellular and intracellular proteins secreted by Candida albicans cells were concentrated using salting-out method and extracted by P0013B RIPA lysis buffer, respectively followed by Western-blotting analysis to detect the expression of extracellular and intracellular phospholipase B1 protein. Results The relative expression of phospholipase B1 mRNA was 0.6173 ± 0.1090 in fluconazole-resistant Candida albicans isolates, significantly higher than in fluconazole-sensitive isolates （0.2653 ± 0.0935, P < 0.05）. Increased expressions of extracellular and intracellular phospholipase B1 protein were noted in fluconazole-resistant Candida albicans isolates compared with fluconazole-sensitive isolates （0.4145 ± 0.2773 vs 0.2720 ± 0.2194, 0.1825 ± 0.1831 vs 0.2653 ± 0.0935, t = 2.703, 3.443, both P < 0.05）. Conclusions The expressions of phospholipase B1 mRNA and protein are elevated in fluconazole-resistant Candida albicans isolates, suggesting that the expression of phospholipase B1 may be associated with the occurrence of antifungal resistance in Candida albicans.

Key words: Candida albicans, lysophospholipase, Drug resistance，fungal