白念珠菌经汉防己甲素作用前后的差异蛋白质组学分析

目的 研究汉防己甲素对白念珠菌蛋白质表达的影响,寻找汉防己甲素作用相关蛋白质.方法 以白念珠菌氟康唑敏感株CA-3为研究对象,常规培养白念珠菌,实验分两组,分别为不加药的CA-3组及加汉防己甲素(终浓度为250 mg/L)的CA-3组,汉防己甲素作用6 h后收集细胞,提取两组样本的总蛋白质,运用固相pH梯度双相凝胶电泳分离,Image Master 2D Platinum软件分析蛋白图谱的差异,以经基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF-MS)对蛋白质进行质谱鉴定.结果 成功建立汉防己甲素作用前后白念珠菌敏感株蛋白质双相凝胶电泳图谱,图像重复性好.图像分析出差异蛋白质26个,最终质谱鉴定出差异蛋白质7个,其中上调蛋白质1个,为Pst1;下调蛋白质6个,分别为Idh1、Asc1、Rps5、Asn1、Asn1、Srb1.结论 汉防己甲素作用于白念珠菌敏感株前后蛋白质有差异,Pst1、Idh1、Asc1、Rps5、Asn1、Asn1、Srb1在汉防己甲素对白念珠菌作用中起重要作用.

Abstract：

Objective To study the effects of tetrandrine on the protein expression profile of C. albicans,and to screen for proteins associated with the effect of tetrandrine. Methods Fluconazole-sensitive C. albicans CA-3 was cultured with or without tetrandrine (250 mg/L) for 6 hours followed by the collection of Candida cells. Total proteins were extracted from these cells and separated by using immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis. Protein spots were detected and analyzed by Image Master 2D Platinum software, and MALDI-TOF/TOF-MS-based method was used to identify these proteins. Results The two-dimensional gel electrophoresis maps of C. albicans proteins before and after the treatment with tetrandrine were successfully obtained with high repeatability. Image analysis revealed a total of 26 differentially expressed protein spots in C. albicans treated with tetrandrine, and 7 differentially expressed proteins were identified by the mass chromatographic analysis, including 1 up-regulated protein (Pst1), and 6 down-regulated proteins (Idh1,Asc1, Rps5, Asn1, Asn1, Srb1 ). Conclusions The expressions of some proteins in fluconazole-sensitive C. albicans experience significant changes after tetrandrine treatment, and Pst1, Idh1, Asc1, Rps5, Asn1, Asn1 and Srb1 may considerab1y contribute to the effects of tetrandrine on C. albicans.

Differential proteomic analysis of fluconazole-sensitive Candida albicans before and after tetrandrine treatment

Objective To study the effects of tetrandrine on the protein expression profile of C. albicans,and to screen for proteins associated with the effect of tetrandrine. Methods Fluconazole-sensitive C. albicans CA-3 was cultured with or without tetrandrine (250 mg/L) for 6 hours followed by the collection of Candida cells. Total proteins were extracted from these cells and separated by using immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis. Protein spots were detected and analyzed by Image Master 2D Platinum software, and MALDI-TOF/TOF-MS-based method was used to identify these proteins. Results The two-dimensional gel electrophoresis maps of C. albicans proteins before and after the treatment with tetrandrine were successfully obtained with high repeatability. Image analysis revealed a total of 26 differentially expressed protein spots in C. albicans treated with tetrandrine, and 7 differentially expressed proteins were identified by the mass chromatographic analysis, including 1 up-regulated protein (Pst1), and 6 down-regulated proteins (Idh1,Asc1, Rps5, Asn1, Asn1, Srb1 ). Conclusions The expressions of some proteins in fluconazole-sensitive C. albicans experience significant changes after tetrandrine treatment, and Pst1, Idh1, Asc1, Rps5, Asn1, Asn1 and Srb1 may considerab1y contribute to the effects of tetrandrine on C. albicans.