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研究艾叶挥发油(Artemisia argyi essential oil,AAEO)诱导白念珠菌凋亡的活性。实验采用微量液基稀释法检测AAEO对白念珠菌(Candida albicans)SC5314的MIC;采用XTT还原法检测AAEO对白念珠菌SC5314代谢的影响;采用流式细胞仪检测AAEO对白念珠菌SC5314细胞内活性氧(reactive oxygen species,ROS)和线粒体膜电位(mitochondrial membrane potential,MMP)水平的影响;采用Annexin-V/PI法测定AAEO对白念珠菌SC5314细胞凋亡率的影响;采用荧光显微镜观察AAEO对白念珠菌SC5314细胞Caspase酶活性的影响;采用DAPI染色法观察AAEO对白念珠菌SC5314细胞核固缩的影响。结果显示,AAEO对白念珠菌SC5314的MIC为0.5 mL·L~(-1);XTT代谢活性显示AAEO对白念珠菌SC5314代谢呈剂量依赖性的抑制;>0.5 mL·L~(-1)的AAEO干预后,白念珠菌SC5314细胞内活性氧水平显著升高、线粒体膜电位显著降低、凋亡比例明显增加、metacaspase活性显著升高、细胞核固缩。AAEO具有诱导白念珠菌细胞凋亡的活性,该作用可能与ROS积累和线粒体损伤相关。
To study the activity of Artemisia argyi essential oil (AAEO) in inducing Candida albicans apoptosis. The MIC of AAEO against Candida albicans SC5314 was detected by micro-liquid-based dilution method. The effect of AAEO on the metabolism of Candida albicans SC5314 was detected by XTT reduction method. The effect of AAEO on the activity of reactive oxygen species reactive oxygen species (ROS) and mitochondrial membrane potential (MMP). The effect of AAEO on the apoptosis rate of Candida albicans SC5314 cells was determined by Annexin-V / PI assay. The effect of AAEO on Candida albicans SC5314 The effect of AAEO on the nuclear condensation of Candida albicans SC5314 cells was observed by DAPI staining. The results showed that the MIC of AAEO against Candida albicans SC5314 was 0.5 mL·L -1. The metabolic activity of XTT showed AAEO inhibited the Candida albicans SC5314 in a dose-dependent manner. AAEO> 0.5 mL·L -1 After intervention, the levels of reactive oxygen species (ROS) in C. albicans SC5314 cells were significantly increased, mitochondrial membrane potential was significantly decreased, the percentage of apoptosis was significantly increased, metacaspase activity was significantly increased, and nuclear condensation was observed. AAEO has the activity of inducing C. albicans cell apoptosis, which may be related to ROS accumulation and mitochondrial damage.