艾叶挥发油诱导白念珠菌凋亡

Activity of essential oil extracted from Artemisia argyi in inducing apoptosis of Candida albicans

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。

Abstract：To explore the activity of essential oil extracted from Artemisia argyi （AAEO） in inducing the apoptosis of Candida albicans SC5314. The effect of AAEO on reactive oxygen species（ROS） and mitochondria membrane potential（MMP） of C. albicans SC5314 was detected by flow cytometry. Phosphatidylserine externalization was observed under fluorescence microscopic with Annexin-V/PI staining at the early stage of apoptosis in C. albicans. Metacaspase activity was observed under fluorescence microscopic with FITC-VAD-FMK staining at the early stage of apoptosis in C. albicans. C. albicans morphology was observed by DAPI nuclear staining and fluorescence microscopy. After intervention with 0.5 mL·L-1 AAEO, apoptosis of C. albicans significantly increased, metacaspase activity increased, nuclear pyknosis and fragmentation, and intracellular ROS were significantly increased, and mitochondrial membrane potential decreased significantly. The certain concentrations of AAEO could induce the apoptosis of C. albicans.