用流式细胞术研究中药对白念珠菌的抗菌作用

Effects of Antifungal Agents from Chinese Herbs on Candida albicans by Flow Cytometry

目的应用流式细胞仪测定中药有效成分小檗碱、黄芩甙、丁香酚和姜黄素对白念珠菌细胞周期的影响。方法将白念珠菌培养在含不同药物浓度的YEPD培养基中,培养48h,流式细胞仪检测细胞生长周期、DNA荧光强度和细胞体积大小。结果4种抗真菌中药有效成分对白念珠菌细胞生长周期有不同程度的影响,随着药物浓度的增高,其处于S-G2-M期的细胞比率越低,亦即细胞分裂受抑制越明显。在含药物培养基中生长的真菌细胞的荧光强度减弱,反映了细胞DNA片段的丢失,并随着药物浓度的升高,荧光强度减弱越明显,反映细胞体积大小、折光度和颗粒度的散点图向下和向左移动,随着药物浓度的升高,这种图形变化越明显。结论中药单体通过抑制细胞分裂发挥抗真菌作用,流式细胞仪可用于抗真菌药物敏感性测定。

Objective To determine the effects of Chinese herbs effective components,i.e.berberine,baicalin,eugenol and curcumin,on cell cycle of Candida albicans by flow cytometry(FCM).Methods After Candida albicans were incubated in YEPD broth which contained different concentration of antifungal agents for48hours,the cell cycle,fluorescent intensity and size of cell volume were detected by FCM.Results The4kinds of Chinese herb monomer could affect the cell cycle of Candida albicans in different range.The ratio of cells in S-G 2 -M period decreased as the agents concentration increased,i.e.the cell division was inhibited.The fluorescent intensity of Candida albicans cells became weak after incubated,which reflected the loss of DNA fragments.The higher concentration,the weaker fluorescent inten-sity was.The size of cell volume,cell diopter and particle size changed more as the agents concentration increased.Con-clusions Chinese herb monomers played the antifungal role through inhibiting cell division.FCM could be used as a method in determining the susceptibility of antifungal agents.