Zhang Xiao-Li, Zhang Ge-Hua, Wang Zhi-Yuan, Guo Hui, Gao Lai-Qiang, Wu Si, Song Yan-Jun, Zhang Hong
The First Affiliated Hospital and Institute of Mycology of Jinan University, Guangzhou, China.
Department of Otolaryngology-Head and Neck Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Pharmazie. 2013 Dec;68(12):951-4.
We previously showed that Adh1p participates in fluconazole (FLC) resistance in Candida albicans through a mechanism that may involve efflux pumps. We also found that the concomitant use of tetrandrine (TET) and FLC provided a synergistic action against C. albicans and that the mechanism of action could be related to inhibition of a drug efflux system. To determine whether Adh1p participates in the synergistic antifungal activity of TET against C. albicans, we performed a comparative proteomic study comparing cells treated with FLC and/or TET in FLC-sensitive CA-3 and untreated control cells. Proteins were analyzed using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), and differentially expressed proteins were identified through matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF/TOF) mass spectrometry. The resulting data were searched against a C. albicans protein database. Our analyses identified six differentially expressed proteins; four (Eno1p, Adh1p, Slb1p, and Tdh1p) were down-regulated, and two (Xyl2p, and Cdc19p) were up-regulated. The Adh1p mRNA levels were consistent with the Adh1p protein levels in all of the groups. The results suggest that Adh1p participates in the synergistic antifungal activity of TET against C. albicans.
我们之前表明，乙醇脱氢酶1（Adh1p）通过一种可能涉及外排泵的机制参与白色念珠菌对氟康唑（FLC）的耐药性。我们还发现，汉防己甲素（TET）与FLC联合使用对白色念珠菌具有协同作用，且作用机制可能与抑制药物外排系统有关。为了确定Adh1p是否参与TET对白色念珠菌的协同抗真菌活性，我们进行了一项比较蛋白质组学研究，比较了在FLC敏感的CA - 3中用FLC和/或TET处理的细胞与未处理的对照细胞。使用二维聚丙烯酰胺凝胶电泳（2 - D PAGE）分析蛋白质，并通过基质辅助激光解吸/电离飞行时间（MALDI TOF/TOF）质谱鉴定差异表达的蛋白质。将所得数据与白色念珠菌蛋白质数据库进行比对。我们的分析鉴定出六种差异表达的蛋白质；四种（烯醇酶1（Eno1p）、Adh1p、Slb1p和甘油醛 - 3 - 磷酸脱氢酶1（Tdh1p））表达下调，两种（木酮糖激酶2（Xyl2p）和细胞周期蛋白19（Cdc19p））表达上调。所有组中Adh1p的mRNA水平与Adh1p蛋白水平一致。结果表明，Adh1p参与TET对白色念珠菌的协同抗真菌活性。
