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中国真菌学杂志 2017, Vol. 12  Issue (1): 13-18.

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白头翁汤正丁醇提取物诱导白念珠菌生物被膜细胞凋亡

施高翔1,2, 汪云霞1,2, 冯鑫1,2, 邵菁1,2, 汪天明1,2, 汪长中1,2   

  1. 1. 安徽中医药大学中西医结合临床学院, 合肥 230038;
    2. 安徽省中医药科学院中西医结合研究所, 合肥 230038
  • 收稿日期:2016-11-23 出版日期:2017-02-28 发布日期:2017-02-28
  • 通讯作者: 汪长中,E-mail:ahwcz63@sina.com E-mail:ahwcz63@sina.com
  • 作者简介:施高翔,男(汉族),硕士,助教.E-mail:ssjlsgx@163.com

Butyl alcohol extract of BaiTouWeng decoction induced apoptosis in Candida albicans biofilms

SHI Gao-xiang1,2, WANG Yun-xia1,2, FENG Xin1,2, SHAO Jing1,2, WANG Tian-ming1,2, WANG Chang-zhong1,2   

  1. 1. School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230038, China;
    2. Institute of Integrated Traditional and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230038, China
  • Received:2016-11-23 Online:2017-02-28 Published:2017-02-28

摘要:

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。

关键词: 白头翁汤正丁醇提取物, 白念珠菌, 生物被膜, 凋亡

Abstract:

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.

Key words: Butyl alcohol extract of Bai Tou Weng decoction, C.albicans, biofilms, apoptosis

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