白头翁汤正丁醇提取物诱导白念珠菌生物被膜细胞凋亡

摘要

Objective To explore the Butyl alcohol extract of Bai Tou Weng decoction effect on apoptosis of Candida albicans biofilms.Methods Using DCFH-DA staining flow cytometry to detect Candida albicans biofilm cells of reactive oxygen species (ROS) level;Using JC-1 staining of mitochondria of Candida albicans biofilm membrane flow cytometry to detect MMP;Phosphatidylserine externalization was recorded by fluorescence microscopic with Annexin V staining at the early stage of apoptosis in C.albicans cells of biofilms;Metacaspase activity was observed by Fluorescence microscopic with FITC-VAD-FMK staining at the early stage of apoptosis in C.albicans cells of biofilms;Observation on Candida albicans biofilm morphology was performed by DAPI nuclear staining and fluorescence microscopy.Results The apoptosis of Candida albicans significantly increased the proportion of more than 128 ug mL-1BAEB treatment,Caspase activity increased,nuclear pyknosis and fragmentation were observed.Intracellular ROS level was significantly increased,mitochondrial membrane potential decreased significantly.Conclusion The Butyl alcohol extract of BaiTou Weng decoction of certain concentrations could induce apoptosis of Candida albicans biofilm cells.%目的 探讨白头翁汤正丁醇提取物(Butyl alcohol extract of Bai Tou Weng decoction,BAEB)对白念珠菌生物被膜细胞凋亡的影响.方法 分别以DCFH-DA染色和JC-1染色,用流式细胞仪检测白念珠菌生物被膜细胞内活性氧(ROS)水平和线粒体膜电位(MMP)变化;Annexin V-FITC/PI染色染色荧光显微镜观察白念珠菌生物被膜细胞磷脂酰丝氨酸(phosphatidylserine,PS)外翻并检测凋亡率;FITC-VAD-FMK染色观察白念珠菌生物被膜细胞metacaspase活性;DAPI染色观察白念珠菌生物被膜细胞核形态.结果 ≥128 μg/mL BAEB处理后白念珠菌生物被膜细胞内活性氧水平显著升高,线粒体膜电位显著降低,PS外翻增加,凋亡率明显升高,metacaspase酶活性显著升高,细胞核出现固缩和碎裂.结论 白头翁汤正丁醇提取物可诱导白念珠菌生物被膜细胞凋亡.