马尔尼菲青霉菌耐氟康唑的蛋白质组初步分析

A preliminary proteome analysis on the fluconazole-resistant strains of Penicillium marneffei

目的探索马尔尼菲青霉菌耐氟康唑的相关蛋白质组。方法将11株菌丝相和酵母相对氟康唑均敏感的马尔尼菲青霉菌菌株在体外含8μg/mL氟康唑的沙氏液基进行耐药诱导培养。E-test法检测各菌株诱导培养前后氟康唑的MIC值,分别筛选菌丝相和酵母相MIC值增加最显著的菌株,通过CM10蛋白质芯片检测菌体药物诱导培养前后蛋白质表达的差异。结果11株菌株的菌丝相和2株菌株的酵母相能耐受8μg/mL氟康唑生长,而且诱导培养7d后菌丝相MIC值显著升高,MIC几何均值由1.22μg/mL上升到55.56μg/mL;菌丝相在耐受氟康唑中16个蛋白质表现为高表达且特异性表达4581.1Da和6109.7Da蛋白质,酵母相则在8μg/mL氟康唑诱导培养下22个蛋白质表现为高表达且特异性表达3575.2Da,8507.0Da和8563.3Da蛋白质。结论马尔尼菲青霉菌菌丝相比酵母相更能耐受氟康唑,该菌对氟康唑的抵抗可能和某些特异性蛋白质相关。

The correlated proteome of Penicillium marneffei resistant to fluconazole was investigated in the present study,in which 11 strains of P.marneffei of both the mycelial and yeast forms sensitive to fluconazole were cultivated in Sabouraud＇s liquid medium containing 8 g fluconazole and the MICs of fluoconazile to P.marneffei before and after inducing cultures were tested by E-test method.The strains of the mycelial and yeast forms showing the most significant increase in MIC value were selected and the protein clip CM10 was used to detect the proteome differences before and after inducing cultures.It was found that 11 strains of the mycelial forms and 2 strains of the yeast forms could tolerate the action of fluconazole and could grow at a drug concentration of 8 μg/mL.Furthermore,the MICs of fluoconazole to the mycelial forms were significantly increased after 7 days of inducing culture and the geometric means of MICs were increased from 1.22 μg /mL to 55.56 μg/mL.After the mycelial forms had been induced to be resistant,16 proteins were highly expressed with specific expression of the 4581.1 Da and 6109.7 Da proteins;whereas 22 resistant yeast form were highly expressed with specific expression of the 3575.2 Da,8507.0 Da and 8563.3 Da proteins.These results suggest that the mycelial form of P.narneffei seems to be more tolerant to the action of fluconazole than the yeast form.The resistance of these organisms to fluconazole may be associated to some specific proteins.