CAJ | 학술논문

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.
목적 연구한방기갑소대백념주균단백질표체적영향,심조한방기갑소작용상관단백질.방법 이백념주균불강서민감주CA-3위연구대상,상규배양백념주균,실험분량조,분별위불가약적CA-3조급가한방기갑소(종농도위250 mg/L)적CA-3조,한방기갑소작용6 h후수집세포,제취량조양본적총단백질,운용고상pH제도쌍상응효전영분리,Image Master 2D Platinum연건분석단백도보적차이,이경기질보조격광해흡전리비행시간천련질보(MALDI-TOF/TOF-MS)대단백질진행질보감정.결과 성공건립한방기갑소작용전후백념주균민감주단백질쌍상응효전영도보,도상중복성호.도상분석출차이단백질26개,최종질보감정출차이단백질7개,기중상조단백질1개,위Pst1;하조단백질6개,분별위Idh1、Asc1、Rps5、Asn1、Asn1、Srb1.결론 한방기갑소작용우백념주균민감주전후단백질유차이,Pst1、Idh1、Asc1、Rps5、Asn1、Asn1、Srb1재한방기갑소대백념주균작용중기중요작용.
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.